Elena Tschumak

INFLUENCE OF FOX GENES ON AGING AND AGING-ASSOCIATED DISEASES

Inhaltsverzeichnis

FOX GENES AFFECT AGING AND RELATED DISEASES

FOXP2 genes have a direct and indirect effect on aging

Influence of other members of Fox-family on aging processes

FOX genes and anti-aging agents

FOXP2 and neuroplasticity

FOXP2 and neurodegeneration

FOXP2 and Alzheimer

Further aging and neurodegeneration relevant FOXP2 targets

Structure and function of FoxP genes is responsible for his function

Natural products and FOXP Targets

Post-stroke FOXP2 expression and discussion on appropriate animal models to study FOXP2 language function

Further literature:

FOX GENES AFFECT AGING AND RELATED DISEASES

FOXP2 genes have a direct and indirect effect on aging

FOX genes include 19 highly conserved, structurally related families from FoxA to FoxS (Sridhar Hannenhalli and Klaus H. Kaestner, 2009). Different publications mentioned the role of FOXgenes in aging processes.

One of the main factors of aging is telomere attrition. Telomeres modulate Rb and p53, that decrease PGC-1α and PGC-1β expression (Sahin and Depinho, 2012) and activates the cyclin-dependent kinase inhibitor p21CIP1.16 H. Shelterin (consists of Tin2, Rap1, TRF1, TRF2, Pot1-TPP1 heterodimer) protects telomers and play a role in mtDNA damage. Telomeric heterochromatin is influenced by lncRNA TERRA via binding to TRF1, TRF2,ORC, HP1 and H3K9me3(Scaffidi et al., 2005; Han and Brunet, 2011).

Tanabe et al. described 2011 in „FOXP2 Promotes the Nuclear Translocation of POT1, but FOXP2 (R553H), Mutation Related to Speech-Language Disorder, Partially Prevents It“, how

FOXP2 influences on POT1 (protection of telomeres 1) as a FOXP2-associated protein. (Tanabe et al. reported protection of nuclear-colocated telomerase.) This gene POT1 as member of the telombin family encodes a nuclear protein involved in telomere maintenance. The loss of its POT1 function induces the cell arrest. The researchers identified that POT1 is alone localized in the cytoplasm but co-localized with FOXP2 and the forkhead domain of FOXP2 in nuclei. FOXP2 with mutated nuclear localization signals as well as R553H mutated forkhead, which is associated with speech-language disorder, prevented the nuclear translocation of POT1. The authors propose FOXP2 as a binding partner for the nuclear translocation of POT1.

According to Multanii and Chang, 2007 „WRN at telomeres: implications for aging and cancer“

POT1 is critically important for telomere extension by telomerase, POT1 functions to negatively regulate telomere length by competing with telomerase for access to the telomeric substrate.

Review Wohlgemuth et al, 2014 showed that FOXP2 also provided the NMDAR-mediated neuronal plasticity affecting MAPKK. It is known that Aging also influences free radicals via Fc-gamma receptor and phagocytosis via p42/p44 MAPK signalling pathways and aging relevant ROS activate inflammatory cascade reaction via JAK/STAT, NF-κB/MAPK.

Molina-Serrano et al. showed 2019 in „Histone Modifications as an Intersection Between Diet and Longevity“ that Lysine 4 on histone H3 can be found in one of three possible methylated forms: mono, di or trimethylated (H3K4me1, -2, and -3).The trimethylated form usually localizes at the promoter of actively transcribed genes (Santos-Rosa et al., 2002), and has been shown to have a strong implication with aging in several model organisms. S. cerevisiae H3K4 methyltransferase complex COMPASS mutants had significantly reduced lifespan (Smith et al., 2008; Ryu et al., 2014; Cruz et al., 2018). “H3K9me3 was only associated with antioxidant genes in females (Strakovsky et al., 2014). Paternal HF diet changed the epigenome in spermatozoa and offspring liver. Specifically, under HF diet, H3K4me was enriched in paternal sperm around the transcriptional start site (TSS) of genes involved in development regulatory processes such as Hoxd11, Hoxd13, Bai3, Foxp2, and Foxa2. In contrast, in the offspring liver, H3K4me was enriched in genes controlling lipid biosynthesis, fatty acid synthesis and the oxidation-reduction process (Terashima et al., 2015).Aging relevant H3K27ac and H3K14ac are acetylated via p300/CBP and its co-activator CREB. cAMP responsive CREB expression is responsible to fasting. So CBP, CREB, CRTC2 and TAF-4 activate together gluconeogenesis genes (Altarejos and Montminy, 2011)” (Molina-Serrano et al., 2019, p.11)

FOXP2 indirectly regulates the FEZF2, the RELN, the FOXO1 and the DYRK1A genes via theRUNX-AUTS2-TBR1cascade In a feedback loop the FEZF2 regulates FOXP2 expression. (Molyneaux et al., 2005) TBR1 also regulates RELN expression (Chen et al., 2002). The RELN in turn regulates FOXO1 expression, which is also indirectly regulated by RUNX2 (Kuhlwilm et al., 2013). It would be of great scientific interest to investigate further whether and to what extent FOXP2 expression via FOXO interaction can influence tumorigenesis and antiaging, as FOXO genes are generally known for their importance in cell cycle and apoptosis. (Kops et al., 2002)

Gascoyne et al. suggested 2015 in „The Forkhead Transcription Factor FOXP2 Is Required for Regulation of p21WAF1/CIP1 in 143B Osteosarcoma Cell Growth Arrest“ that “FOXP2 expression could be induced by MAPK pathway inhibition in growth-arrested 143B cells, but not in traditional cell line models of osteoblast differentiation (MG-63, C2C12, MC3T3-E1)” (Gascoyne et al., 2015, p.1) They studied a model in which transient upregulation of Foxp2 in pre-osteoblast mesenchymal cells regulates a p21-dependent growth arrest checkpoint and identified that Foxp2 expression is not neuronally restricted and is linked to regulation of the cell cycle via various mechanisms: p53 target gene cyclin-dependent kinase inhibitor p21, is as a primary controller of multiple diverse cellular pathways, e.g. neuronal differentiation, growth arrest of pre-osteoblast type 143B osteosarcoma cells etc. They reported that regarding the RUNX-2-dependent pathway for FOXP the bone-specific isoform of RUNX2, which these cells lack, is not a requirement for FOXP2 function and in “developing murine mesenchymal cells the loss of growth factor signalling upon entry to the periosteum might up-regulate Foxp2. Expression of Foxp2 during both osteoblast and some neuronal development appears to be transient and this is also observed for other family members”(Gascoyne et al., 2015, p.13), considered potential connections between FOXP2 and p53 pathways. “Normal osteoblast development is compromised in bone metastases of solid tumours and in bone malignancies such as multiple myeloma, and this characterisation of FOXP2 growth arrest function in a disease context may identify novel malignant pathways. Furthermore, FOXP2 status in osteosarcoma may provide information regarding the stage of developmental block, with potential clinical significance. Direct connection of FOXP2 to the mutated p53 pathway in osteosarcoma, via a common target p21/CDKN1A, is also likely to have implications for understanding osteosarcoma biology.” (Gascoyne et al., 2015, p.14)

Also the review of Wohlgemuth et al., 2014 showed that FOXP2 also provided the NMDAR-mediated neuronal plasticity affecting MAPKK and tyrosine phosphatase. The MARK is a RAS Downstream Effector. RAS is a GTPase that indirectly interacts with oncological relevant p21 and p53 indirectly. So FOXP2 can act alone or in combination with other members of its or other gene families. Its own expression can be influenced both by the cooperation partners and by the feedback loops or by other interactions. These innumerable tissue-specific regulation possibilities allow a fine-tuning and a rapid adaptation to ever-changing environmental conditions which need to be studied more closely.

Influence of other members of Fox-family on aging processes

According to Ni et al. 2012, Ribarič 2012 Foxo plays an important epigenetic role in aging and Hansen et al, 2013 describe DAF/FOXO effect on longevity. Steroid hormone dehydrogenases, cytochrome P450s and others aging relevant steroid hormone signalling pathways are influenced not only by LIPL-4 and TOR regulated autophagy but also by daf-16/FOXO (Lapierre et al., 2011).In C. Elegans daf-16/FOXO affects aging and tumor growth. (Pinkston-Gosse and Kenyon, 2007) Experiments with germline-less C. Elegans showed that FOXO3A effects NAD-dependent protein deacetylases and ADP-ribosyl transferases activity of aging relevant sirtuin (Oberdoerffer et al., 2008; Wang et al., 2008) e.g. with the help of histone H3K9 deacetylation, glucose homeostasis, genomic stability and via subunit RelA NF-κB regulating SIRT6 (Kanfi et al., 2010; Kawahara et al., 2009; Zhong et al., 2010). SIRT6 also effects telomers. (Michishita et al., 2008; Mostoslavsky et al., 2006; Tennen & Chua, 2011) AMPK and SIRT1 also directly influence PGC-1α via deacetylation and phosphorylation). Aging is associated with upregulation of chaperones and proteases level via misfolded proteins also appeared by Alzheimer’s and Parkinson’s disease,(Bernales et al., 2012) and UPRmt. UPRmt as well as FOXO signalling can be activated by NAD+ and increase longevity (Mouchiroud et al., 2013) Lon protease is another important for degradation of oxidized proteins within the mitochondrial matrix aging factor (Ngo et al., 2013, 2009; Bota et al, 2002)

FOXO3 (like mTOR and AMPK) effects aging relevant mitophagy (Rodriguez-Hernandez et al., 2009). Kim et al.,2014 describe how phosphorylation, acetylation, or ubiquitination of FoxO-genes effect histones and chromatin and change this way ROS-level. Therefore FoxO4 shows negativ effect on PI3K/Akt pathways (Karger et al. 2009) as well as on MAFbx and MURF1 in muscle aging (Clavel et al. 2006). Oxidative stress leads to FoxO phosphorylation via Akt and its transport from the nucleus to the cytoplasm. FoxO3 and FoxO4 deacetylation by Sirt1 (Jian et al. 2011) and its upregulation via catalase (Fukuoka et al. 2003; Brunet et al. 1999) play an important role in aging. FoxO1 also regulates apoptosis, DNA repair and cell cycle arrest (Yamaza et al.2010; McLoughlin, 2009; Ma et al., 2016), but also plays a key role in stem cell pluripotency. (Zhang et al., 2011). FoxO6 effects gastric carcinoma (Kim et al. 2011) and together with PGC-1a influences oxidative metabolism in skeletal muscle. Chung et al.,2013 reported that FoxO6 and PGC-1a form a regulatory loop to regulate oxidative metabolism in skeletal muscle. In addition, Foxoa2 plays a potential role in sepsis (Berg et al., 2006) and in asthmatic mucus secretion (Park et al., 2009).

An interesting aging approach (connected with adjacent to Paneth cells and crypts) are stem cells and their regulation e.g. via cancer relevant Lrig1, Wnt, Olfm4, Hopx, p57, Sox9, Tert, Bmi1, β-catenin, Ascl2, Lgr5, Myc, Ephb2, CD44, PPAR and SMAD signalling (Nalapareddy et al., 2017; Akunuru, and Geiger, 2016) Deficiencies in DNA damage repair limits the function of haematopoietic stem cells with age. (Rossi et al., 2007) Low ISC and increased ROS levels relate to Foxo1, Foxo3a and Foxo4 activity. (Tothova et al., 2007; Miyamoto et al., 2007)

But FOXO like CoQ and PGC-1α also influences electron transport changes contribute to aging effects such as cardiac failure. (Rosca et al., 2008; Frenzel et al., 2010; López-Lluch et al.,2010; Houtkooper et al., 2010) Shijin et al. mentioned 2016 in „An Update on inflammaging: Mechanisms, Prevention, and Treatment“ the influence of inflammatory cytokines on lymphocytes. Type I and type II cytokines effect CD4+ T lymphocytes (Alberti et al., 2006) and CD8+ and CD4+ T lymphocytes (Franceschi et al., 2000). Especially role on imflamaging play IL-1, IL-6, TNF-α and PGE2 (Bruunsgaard et al., 2003; Cesari et al., 2004). Aging is associated with IGF-1 down- and IL-6- and TNF-α- upregulation (Lio et al., 2003) IL-1, IL-6, and IL-8 are activated via NF-κB signalling pathway (Bartek et al., 2008).

Different publications mentioned that FOX genes are an important factor in the development of tumors (Nishimura et al.,1998; Wang et al., 2015; Tian et al., 2015; Lo et al., 2016,2018; Lu et al., 2017; Nik et al., 2013; Shi et al., 2016; Yu et al., 2017; Dou et al., 2017; Milewski et al., 2017; Cai et al., 2015; Xian et al., 2018; Herrero and Gitton, 2018; Toma et al., 2011; Zhang et al., 2012; Rousso et al., 2012; Howarth et al., 2008; Teufel et al., 2003; Li and Tucker, 1993; Ji et al., 2016; Myatt et al., 2007) The FOXO family and the transcription factor NF-kB influence the upstream protein kinase B e.g. in connection with the receptors of the Trk family (e.g. the Neurotrophin). Inhibition of NF-κB signalling, NLRP3 inflammasome and other pro-inflammatory pathways or hormonal treatments eventually can restore aging relevant Progerin level, elevated by telomeres disfunction. (Osorio et al., 2010; Cao et al., 2011) In the same time senescence is accompanied by increased IL-1ß level and activated tumor necrosis factor and interferons production. (Review López-Otín, 2013; Green et al., 2011; Salminen et al., 2012, 2019; Adler et al.,2007). The Hsp90 and the IκB-Kinase (IKK) inactivate the NF-κB pathway and inhibit autophagy via induction a cell signalling switch from autophagy to apoptosis in tumor cells downregulation and of Beclin 1 expression. (Jiang et al, 2011) NF-κB also reduces GnRH production (Zhang et al., 2013) and affects this way muscle weakness, reduced neurogenesis, bone fragility and skin atrophy. Further NF-κB decreases H3K27me3 level via H3K27me3 demethylase (Lauren et al., 2016; De Santa et al., 2007). BCL2, transcriptionally regulated by nuclear factor-kappa B, affects cell shrinkage (Chakraborty et al., 2015; Lambie1 and Conradt, 2016) and acts directly on apoptosis-activating BH3. Reqmi et al. discovered that BCL2-related proteins also regulate mitochondrial dynamics using dynamin-related GTPases. (Lüpertz, 2008; Hornstein et al., 2013)

Hsp90 protects 20S proteasome from oxidative damage inactivation (Höhn et al., 2017; Conconi et al., 1998) and helps to fold oncogenic proteins e.g. of p53 (Saibi et al., 2013). Aging relevant Hsp70 is activated with the help of HSF-1deacetylation by SIRT1 (Westerheide et al., 2009). Aging dependent changes in the amount of sensitive to diet and exercise NAD+ (Cantó et al., 2015) can affect the activity of sirtuins. SIRT1 activity can be decreased via NAD+ level, which also increases activity of the HIF-1α transcription factor resulting in changed in oxidative phosphorylation and mitochondrial dysfunction. (Gomes et al., 2012, 2013; Greer et al., 2007) SIRT1, p53 and HIF-1α in turn effect NAD+ level. Also, AMPK, which inhibits insulin/IGF-1/mTOR, cooperate with SIRT1 to create new mitochondria with the help of PGC-1 and p53. In the same time mitochondria influences TCA cycle. (Salminen et al., 2014) Tollefsbol described 2014 the connection between caloric restriction and longevity. Schultz and Sinclair described 2016 that intestinal stem cells can renewal via BST1 (converts NAD+ cADPR) (Yilmaz et al., 2012 These cells express Notch ligand delta Dl and ESG.

Low protein-high carbohydrate diet influences energy level via enhancing FGF21 expression and in the same time reduction of mTOR activity. This increases Brain-derived neurotrophic factor expression (Zaptan et al., 2015) and influences neural precursor cells (Marosi and Mattson, 2014; Vivar and van Praag, 2013) Neural progenitor cells are also positively influenced by nutrient-sensing protein FOXO3 ( Renault et al., 2009; Devin et al., 2016; Renault et al., 2009) Also grey matter volume of subcortical regions is positively affected this way. (Colman et al., 2009)

Low caloric intake has also positive effect on SIRT1 activity which leads to dendritic outgrowth and plasticity and low inflammatory cytokine activity. (Maalouf et al., 2009) Ohkura et al., 2010 described how FOXO interacts with the sirtuin, which is relevant to aging and cancer. Sirtuin in turn decreases apoptosis via FOXO or BAX. (Dang et al., 2009; Dang et al., 2009; Martins et al., 2015) According to „CYB5R3: a key player in aerobic metabolism and aging?“ (de Cabo et al. 2010) coenzyme Q (CoQ) helps to reduce NADH-level in mitochondria and regulate this way NAD+-dependent enzymes e.g., sirtuins. So, cytochrome b5 coding reductase CYB5R needs NADH and CoQ for its function.(Villalba et al., 1995) Also low calories intake enhanced aerobic metabolism. This needs cytosolic cooperation of N-myristoylated CYB5R3 (a component of P-450 mediated hydroxylation of drugs and steroid hormones (Passon and Hultquist, 1972), fatty acid elongation (Keyes and Cinti ,1980) and cholesterol biosynthesis (Reddy et al, 1977)) and SIRT1. SIRT1 in turn needs lysine acetylation for this cooperation. It modifies CYB5R3 activity and regulates this way the cytosolic NAD+/NADH level.

The activity of FOXO/DAF-16 is positively and negatively regulated by different molecular players (e.g., the insulin/IGF signalling pathway and the nutrient sensor AMPK) and stresses (e.g., oxidative and heat stress). (Eijkelenboom and Burgering, 2013; Salih and Brunet, 2008; Dervis and al., 2008) Sirtuins and H2S induced AMPK and IIS (influences FOXO and mTOR (effected by GH)) have opposite actions. (Barzilai et al., 2012; Fontana et al., 2010; Kenyon, 2010, 2005; Blagosklonny, 2006; Kapahi et al., 2010; Stanfelet al., 2009; Polak and Hall, 2015; Moskalevet al., 2014) FOXO also effects detoxification enzymes MnSOD and GADD45. (Kops et al., 2002; Nemoto and Finkel, 2002)

Apolipoprotein E4 (apoE4) and FOXO are not only associated with quick cell aging (Aksenov et al., 2001; Martins, et al., 2015) but also with Alzheimer’s disease (Sando et al., 2008; DiLoreto & Murphy, 2015).

Further aging factors are rapamycin target S6K activator and 4EBP1 inhibiter mTORC1 and cytoskeletal relevant TORC2 protein kinases (Zoncu et al., 2011),

AMP/ATP ratio and metformin activated AMPK, insulin and IGF1 influenced (FOXO) , which in turn activates MnSOD.

FOX genes and anti-aging agents

Aging relevant SIRT2 deacetylase can be regulated via Resveratrol. Resveratrol is known for his positive affect on Alzheimer disease, on neuroendocrine tumors, on multiple myeloma, on follicular lymphoma, on colon-ca etc. Like EGCG and alpha-M it inhibits PI3K/PTEN/Akt/mTORC1 and WNT/beta-catenin pathway. Resveratrol effects c-Myc, MMP-7 and SIRT1activity and increases SLUG-, vimentin- and NF-kappa-B level. NF-kappa-B level in turn influences caspase-3, MMP-9- and CXCR4- level as well as EMT-level via TGF-beta/SMAD. TGF-beta/SMAD regulates SNAIL/E-cadherin level. ( Ji et al., 2015)

Resveratrol also increases PARP-1 and AMPK level (which in turn decreases MDR1 and CREB phosphorylation level (Wang et al., 2015), E-cadherin-level and apoptosis. It decreases cell migration, invasion and proliferation. Resveratrol interacts with type II topoisomerase and causes double strand DNA breaks and also influences TP53 via ATM. (Demoulin et al., 2015) Together with mitomycin it effects C p21it level and shows an anti-proliferative effect. (Ali et al., 2014) Resveratrol increases the expression of miR-34 which resulted in the decreased expression of E2F3 and SIRT1 (Kumazaki et al., 2013) Tumor suppressor Klotho gene have also aging relevant effect. It increases SIRT1 level (Melatonin shows similar effects) and activates FOXO3 phosphorylation via PI3K/PTEN/Akt pathway. Resveratrol has positive effect on liver cancer and macular degeneration (via VEGF-A and VEGF-C negative regulation), on cardiovascular diseases, on endometriosis, on PCOS, on dyslipidaemia, on diabetes, on chronic renal insufficiency, on Friedreich Ataxia, on Huntington disease and on brain ca via TP53 and p21, 1A/1B light chain 3B (LC3-II), Atg5, beclin-1 and NANOG Cip1 etc. Resveratrol inhibits cyclooxygenase activity, CYP A1 metabolism and influences aging relevant TNFRSF6,TNF-alpha, HIF-1alpha, VEGF, NF-kappaB activity, FAS/FAS-ligand, TP53, FAS-L = CD95, IL-17 but also apoptosis of activated T cells interleukin 17. In head and neck cancer Resveratrol decreases ALDH1, CD44 , HNC-TICs, EMT, NANOG, NESTIN and OCT4 level (Hu et al., 2012) In leukaemia it decreases pLKB1level via SIRT1 and STK11 (Peng et al., 2015) Antiaging phytochemicals are Stilbenoid Resveratrol from Veratrum, which among others influences caspase-1 (Yang et al., 2014; Pietrocola et al., 2012) as well as Catechins and EGCG from Green, which also influence Beclin 1 (Yang et al., 2013; Yang, 2008; Fan et al, 2014) and Camellia sinensis. which also positively influences Helicobacter pylori-triggered caspase-1 signalling pathway.

Not only Resveratrol but also Andrographolide and Parthenolide effect NF-kappaB (Gunn et al., 2011) Further antiaging products are e.g. Iberis amara, which effects apoptosis and ROS ( Weidner et al., 2016) and Beta-carotene, which interact with FOXP2 and piperine, which like Vitamin A kill CSCs. ( Scarpa et al., 2015) Omega-3 polyunsaturated fatty acids negatively influences PI3K/PTEN/Akt/mTORC1 signalling and effects apoptosis (Vasudevan et al., 2014) According to (Rafael de Cabo et al., 2014) caloric restriction influences FGF21, TOR/S6K pathways, insulin, SIRT3, mitochondrial acetyl proteome (Hebert et al., 2013) and metformin - ATP Level in mitochondria, SKN-1/Nrf (Berstein, 2012) (in the same time in worms it lowers ATP levels (Dillin et al., 2002; Lee et al., 2003) possible via mutations in the mitochondrial leucyl-tRNA synthetase gene (Lee et al., 2003),Atg5 (Pyo et al., 2013), TOR signalling, p53 (Jia and Levine, 2007; Tavernarakis et al., 2008).Also acetylproteome, spermidine and sirtuin, which influences autophagy, histone acetyltransferase like resveratrol effects histone deacetylase (sirtuin) (Morselli et al., 2010) and Atg5 (Morselli et al., 2011).SIRT3 is positively affected by dietary restriction via deacetylation of mitochondrial proteins (Someya et al., 2010) and also can be upregulated by Resveratrol. ROS activates hypoxia-inducible factor 1 (HIF-1) and (AMP)-activated protein kinase (AMPK) and affects transcription factors NRF2/SKN-1 and p53/CEP-1 (Chang et al., 2015; Ventura et al., 2009 ). AMPK activates PGC1α , which regulates mitochondrial respiration and detoxification( Liang and Ward, 2006 ). Low PGC1α level effects antioxidant NO (Borniquel et al., 2006 ) which also contribute to proper memory functions. CR also positively influences Hormesis (Calabrese et al., 1987).and anti-stress transcription factors e.g. Gis1, Msn2/Msn4 and Rim15 in yeast and FOXO in mammals.(Wang et al., 2011) In flies it needs help of Complex I and IV (Zid et al., 2009), d4E-BP (Zid et al., 2009) and catalase (Schriner et al., 2005).

Several reports illustrated that Metformin activates AMP-activated protein kinase (Zhou et al., 2001) and inhibits tyrosine phosphates activity (Holland et al., 2004) . and mTORC1 signalling (Kalender et al., 2010)

Campisi showed in her review „Senescent Cells, Tumor Suppression, and Organismal Aging: Good Citizens, Bad Neighbors25 February 2005“ p53 and RB tumor suppressor proteins as aging relevant agents with anticancer effect. Senescence is associated with RAS-RAF-MEK signalling cascade. Different senescence signals can converge (Bringold and Serrano, 2000; Lundberg et al., 2000; Narita and Lowe, 2004), so p53 can be influenced by telomeres (d’Adda di Fagagna et al., 2003, Takai et al., 2003) and RAS (Serrano et al., 1997; Ferbeyre et al., 200;, Pearson et al., 2000), RAS also upregulates pRB (responsible for repressive heterochromatin at loci containing transcription factor E2F) and effects p16 (Beausejour et al., 2003;Wright and Shay, 200;, Collins and Sedivy, 2003; Ben-Porath and Weinberg, 2004; Harvey et al., 1993; Smogorzewska and de Lange, 2002) and plays a role in oxygen toxicity during replicative senescence (Parrinello et al., 2003), p16, p53 pathway, which overlap with oncogenic RAS (Beausejour et al., 2003; Brookes et al., 2002; Huot et al., 2002), but also with ROS (Irani et al., 1997), IGF1 signalling pathway, ARF, MDM2 (Krishnamurthy et al., 2004), p16 transcriptional activator Ets-1 (Ohtani et al., 2001), correlated with p16 expression (Krishnamurthy et al., 2004). It is possibly due to increased sensitivity of the INK4a/ARF locus to transcriptional activation.

According to Blagosklonny, 2012 sirtuins negatively regulate rDNA recombination (Kaeberlein et al., 1999) and number of extrachromosomal rDNA circles. (Stinclair and Guarente, 1997) Sir.2 improves FOXO ortholog, DAF-16 function in worms possibly via IIS (Tissenbaum et al., 2001) and in flies via Rpd3 deacetylation. (Rogina and Helfand, 2004)

Rapamycin inhibits mTOR and rejuvenates cardiac and skeletal muscles via increasing NAD+ level (Cantó et al., 2012)and improves aged immune system (Mannick et al., 2014). It also activates Sirtuin via STACs. STACs are impaired by TGF-β, the levels of which increases during aging in mouse and human sera. Another possibly aging relevant effect of Rapamycin is inhibition of S6K1 (Selman et al., 2009 ). Similar effect on increases longevity has nutrient supplementation with spermidine and polyamine-production of gut flora in mice (Tofalo et al., 2019; Matsumoto et al., 2011, 2007)

According to „Six plant extracts delay yeast chronological aging through different signalling pathways“ Lutchman et al., 2016 Cimicifuga via SNF1 (TORC1 inhibition), Ginkgo biloba via PKA inhibition, Valeriana officinalis L. via PKA pathway, Apium graveolens L. and Salix alba via PKH1/2-sensitive form of Sch9 inhibition as well as Passiflora incarnata L, caffeine, myriocin, spermidine, cryptotanshinone, rapamycin, lithocholic acid, resveratrol and methionine sulfoxide positively effects Saccharomyces cerevisiae aging . (de Cabo et al., 2014; Eisenberget al., 2009 Fontanaet al., 2010; Goldberget al., 2010, 2009; Hubbard and Sinclair, 2014; Kaeberlein, 2010; Leonov et al., 2015; Minois et al., 2011; Huang et al., 2014; Morselli et al., 2009; Arlia-Ciommo et al., 2014; Burstein et al., 2012; Lutchman et al., 2016)

SNF5 component SWI/SNF acts in opposite to polycomb-mediated PcG silencing in Drosophila and to p16INK4a silencing in mammals, leads to increased p16 level and is decreased in cancer cells. (Oruetxebarria et al., 2004)

Further natural aging relevant compounds describe McCubrey et al. 2017 in “Effects of resveratrol, curcumin, berberine and other nutraceuticals on aging, cancer development, cancer stem cells and microRNAs”

Rapamycin also protects again Alzheimer's disease, Parkinson and cardiomyopathy via mTOR (Cai Z and Yan LJ, 2013; Bitto et al., 2015). TOR overlaps with metformin, which increases insulin sensitivity, activates AMPK and inhibits IGF-1 signalling65. H2Ss is also a goal of metformin. (Anisimov, 2010; Awartani, 2002). Nestler et al., 2002 describe that Biguanide have positive effect on ovulation and gynaecological diseases. It also improves immune response (Dilman ,1994), has a positive effect on N-nitrosobis-(2-oxopropyl) induced pancreatic cancer malignancies (Schneider, 2001), inhibits lung carcinogenesis induced by tobacco carcinogen (Memmott et al., 2010) and benzo(a)pyrene-induced skin and cervico-vaginal carcinogenesis in mice as well as on melatonin level (Deriabina et al., 2010) .Phenformin improves immunity and inhibits carcinogenesis in mice (Vinnitski and Iakumenko, 1981) and can inhibit carcinogenesis induced by X-rays in rats. (Anisimov et al., 1982) Diabenol® showed anticancerogenic effects, eg. in colon cancer, improved oestrous function, decreased the size of mammary adenocarcinoma metastases into the lung, multiplicity of all colon tumors in mice. (Popovich et al., 2005)

Metformin as buformin positively effect aging relevant glycation end products (Kiho et al., 2005; Diamanti-Kandarakis et al., 2007) Therefore pentosidine is an aging-marker (Ulrich, 2001)

According to Anisimov et al., 2013; Perridon et al., 2016 ; Wu et al., 2012 various H2S upregulates SIRT, which can inhibit human colon adenocarcinoma. H2S also improves mitochondrial function and antioxidants level.( Kimura et al., 2010, 2004) It happens via ROS-scavengers (Sun et al., 2012) and S-sulfhydration of p66Shc (Xie et al., 2014) glutathione peroxidase, superoxide dismutase. H2S stabilize and activates p21 (van Deursen et al., 2014) It also effects anti-oxidative transcription factor Nrf2 activation via S-sulfhydration of Keap1 (Yang et al., 2013), induced the S-sulfhydration of MEK1 and protects against human platelet aggregation in vitro ( Gao et al., 2015) According to Review „The role of hydro gen sulphide in aging and age-related pathologies“ Bernard et al., 2016, H2S has positive effects of on genome stability via increasing MEK1 S-sulfhydration, ERK1/2 and PARP-1 activity leading to the activation of DNA damage repair mechanisms and protection from cellular senescence.(Zhao et al., 2014)

The CSE/H2S pathway is important in genome stability and cell proliferation due to downregulation of ERK1/2 activity: its inhibition in hepatoma cells decreases their proliferation and increases ROS production, mitochondrial disruption, DNA damage and apoptosis. H2S inhibition activates p53, p21, Bax and other pro-apoptotic genes (Pan et al., 2014) and protects against oxidative damage of Alzheimer's patients . H2S also effects telomere maintenance and influences expression and activity of SIRT1 (López‐Otín et al., 2013; Zhang et al., 2014; Li et al., 2014), IL-6, TNF-α (Rios et al., 2015; Zhang et al., 2013) and methylation via CSE/H2S. (Li et al., 2015; Kamatet all., 2015) NaHS inhibits glycation in humans. (Houtkooper et al., 2010)

H2S impairs the insulin/IGF-1 pathway in neuroblastoma (Liu et al., 2013) and blocks nutrient-sensing mTOR pathway and protein aggregations. (Talaei et al., 2013) H2S inhibits FOXO1 and FOXO3 phosphorylation and is an endogenous regulator of PTEN, the main antagonist of the PI3K-Akt axis in the insulin/IGF-1 signal pathway. The insulin/IGF1 signalling pathway and the FOXO transcription factors modulate the expression of pro-longevity genes (Lee et al., 2003 Oh et al., 2006 Murphy et al., 2003) Marjolein et al., 2017 also demonstrated Foxa2- FOXO4-p53 interaction and its role in cell aging. Its overexpression has been reported in metastases of prostate cancer (Mirosevich et al., 2006) and in colorectal carcinoma (Lehner et al., 2007), while in thyroid cancer cells Foxa2 suppression has been observed. (Akag et al., 2008)

Other FOX family genes are involved in apoptosis, cell invasion and migration, cellular development, metabolism, cell proliferation and development. (Myatt et al., 2007)

Their mutations can lead to cellular cancerization. (Kalin et al., 2006), (Paik et al., 2007)

Several other publications also confirmed the relationship between FoxP expression and cell proliferative, oncological and inflammatory processes as well as the relationship between other FOX genes and vitiligo (FOXD3), ptosis (FOXL2), adipose disease (FoxY2) (Zhou et al. 2018) and lymphoedema-associated autosomal dominant lymphedema-distichiasis syndrome (Fang et al., 2000).

FOXG1 plays a role in apoptosis but is also expressed in the placenta. (Entrez Gene: FOXG1B forkhead box G1B FoxF2).

Among other, FOXJ1 regulates the immune system. (Lin et al., 2005)

FOXA2 influences aging relevant changes in nuclear lamina and increases the lamin B1- and aberrant prelamin A isoform (called Progerin)-level (Bochkis et al., 2014; Scaffidi et al., 2006).

FOXM1B, the „fountain of youth gene“, stimulates tissue growth. (Laoukili et al., 2005; Wierstra and Alves, 2007) and caloric restriction effect on aging can happen via FOXL2 and Foxa2 (hepatocyte nuclear factor 3-beta, HNF3b).

Foxa2 among others effects dopaminergic neurons (its mutation contributes to locomotor deficits in animals that remember the human Parkinson's disease (Kittappa et al., 2007; Arenas, 2008; Lin et al., 2009). It also influences sleep through melanin-concentrating hormone (MCH) and orexin (which also plays a role in alcohol dependence).

FOXP3 importance for carcinogenesis The tumor suppressor gene p53 increases Foxp3 expression by binding directly to the its promoter region of the conserved non-coding DNA sequence CNS-2. (Kawashima et al., 2013) P53 may not only play its immunosuppressive but also its anticarcinogenic role through FOXP3 activation. As already mentioned, protooncogene p53 plays an important role in aging and aging relevant diseases.(Reinhardt And Schumacher, 2012; Carrasco‐Garcia et al., 2017; Nicolai et al., 2015) p53 controls ROS in connection with expression of GLS2 via Tap63. p53 plays a role in apoptosis (Li et al., 2014) especially with the help of mitochondrial pathway of PUMA (Westphal et al., 2014; Yu et al., 2003) and with the help of pro-apoptotic proteins.(Zahran et al., 2014) e.g., via NOXA (Cuadrado Cuadrado et al.,2007) p53 also activates tumor metastasis via SNAI2 (Wu et al., 2005). Further ATP-dependent chromatin remodeler p53 interacts with CSB, which is important for DNA repair.(Lake et al., 2011) It would be interesting to investigate whether and to what extent FOXP3 is involved in a variety of carcinogenic diseases with T inflammatory response.

According to the 28th annual meeting of the „German Association for the Study of the Liver“ (GASL) almost every 5th cancer is caused by chronic inflammation. For example, H. pylori produces the toxic ammonia and vacuolating cytotoxin A, they are demaging gastric tissue. This activates cytokines, which in turn activate neutrophilic granulocytes, T cells and macrophages. H. Pylori attempts to induce apoptosis of macrophages and T cells. This dysregulation of tumor-protective immune processes leads to the development of gastric carcinoma. (Liliane Sygulla, 1998) Human herpesvirus-8 (HHV-8) also involves vascular endothelial growth factor interleukin-6 and VEGF-like genes, which promotes tumorigenesis. (Masood et al., 2002) Human T-lymphotropic virus 1 (HTLV-1) can induce tropical spastic paraparesis or adult T-cell leukaemia (ATL) in which T cells respond independently of the external growth signals. (Liesz et al., 2012)

Gerontologic medicine also tries to prevent the inflammations on a cellular level. For example: many natural anti-aging agents protect against inflammatory process (e.g. curcumin and cayenne pepper) (Aggarwal, 2010) or to influence growth processes (polyphenols, e.g. epigallocatechin EGCG from green tea or procyanidin from apples). (Lombardo et al, 2015)

Posselt's et al. working group investigated 2016 in their work „Spatial distribution of FoxP3 + and CD8 + tumor infiltrating T cells reflects their functional activity“ FoxP + and CD8 + distribution in regulatory and cytotoxic T cells. According to their results the cytotoxic T cells play the key role in the immunological cancer response and offer promising new ways in the fight against cancer. Further results were reported by Schmidt et al. (2011), Lin et al. (2015), Gerber et al. (2014), Li et al. (2016) and Balbaco et al. (2014). These studies showed that T-cell activity were decreased by the PD-L1 and increased by the CTLA-4 antibodies. Both molecules are influenced by the FOXP3 gene. Jim Allison took advantage of this feature in the development of the cancer immunotherapeutic Ipilipumab 2008. FOXP3 interaction with other chemotherapeutic agents has been reported by Takada et al. (2018), Tang et al. (2008) and Landoire et al. (2016). Interestingly, Programmed Cell Death Ligand 1 correlated with FOXP3 expression. (Enkhbat et al., 2018)

The role of l n RNA in aging

Long noncoding RNAs are also aging relevant (Kim et al., 2016) e.g. via phosphatase and tensin homolog PTEN. ln RNA represses expression of the age related tumor-suppressor gene via its interaction with DNMT3a. lncRNA TARID induces promoter demethylation of TCF21. lncRNAs 7SL and MEG3, which is of importance in carcinogen processes (Li et al., 2018), decreased cell proliferation via MDM2 and the tumor suppressor p53 inhibition (Zhou et al, 2007) is decreased in liver, bladder cancer and in pituitary cancer (Braconi et al. 2011; Zhang et al. 2003; Ying et al., 2013) It influences autophagy inversely in age related gastric cancer. LncRNA HULC triggers autophagy direct via stabilizing Sirt1 and attenuates the chemosensitivity of HCC cells (Xiong et al., 2017) As others lncRNAs (Reviewed Yanget al., 2014) HULC influences tumorigenesis in liver and colorectal cancer. (Panzitet al., 2007; Wang et al., 2010)

ncRNAs also modulate protein transcription indirectly via microRNAs. E.g., lncRNAs such as linc-MD1 and lincRNA-RoR change the level of another mRNAs. Grammatikakis

et al., 2014; Cesanaet al., 2011; Wanget al., 2013) In the same time Uchl1 lncRNA may influence senescence directly via p14ARF and p53 and its repressor MDM2 (via proteasome). NF90, required for hypoxia-induced cancer cell invasion (Petrovics et al, 2004; Yang et al., 2013) also effects lnc RNA decrease and is also cancer and apoptosis relevant , because it reduces p21, P53, caspase 7, PARP and PCGEM1 level. (Fu et al., 2006) Cancer relevant lncRNA PVT1 is downregulated via MYC, which is also associated with proliferation and apoptosis of breast and ovarian cancer (Carramusaet al., 2007).

According to Kim et al. „Long noncoding RNAs in Diseases of Aging“, 2016 MALAT1 interact with TP53 and decreases with aging, based on changing serine/arginine (SR) splicing factors level. (Abdelmohsen al. 2013 Tripathi et al.; Tripathi et al., 2010; Melzeret al., 2013) Its level is increased lung and liver carcinoma (Qi et al., 2013; Luo et al., 2006) and influences muscle aging.

ANRIL influences cell cycle as well senescence via upregulation of the tumor suppressor gene p15 (INK4B) (Latrese et al, 2000; Kotake et al., 2011; Peters 2008) and plays a role in breast and another carcinoma, e.g. in gastric, prostate cancer cells and non-small cell lung cancer cell via KLF2 and p21 downregulation (Yap et al., 2010; Nie et al., 2014;1:268–277; Zhang et al., 2014; Hannou et al., 2015; Pasmant et al., 2011) and disturbs miR-99 and miR-449a. LncRNA PINT interacts with PRC2 complex via SUZ12, with plays a role in histone modification and cancer. (Marin-Bejar et al., 2013)

HOTAIR also influences senescence and cancer, e.g., breast tumors as well as endometrial, colorectal, cervical carcinomas and metastasis via H3K27me3 changes and chromatin modifiers, e.g. PRC2. (Kogo et al., 2011; Zhang et al., 2013; Heet al, 2014; Huanget al., 2014; Gupta et al., 2010)

The lncRNA XIST is decreased in breast cancer (Vincent-Salomon et al., 2007) and correlates with Taxol sensitivity. (Huang et al., 2002)

a 200-nt-long ncRNA BCYRN1 is associated with AD b and aging (Mus et al., 2007) but also with breast (Iacoangeli et al. 2004), parotid, tongue, oesophagus, lung, cervix and ovary cancer (Chen et al, 1997).

GAS5 lncRNA suppressed GR and induced apoptosis (Mourtada-Maarabouni et al., 2009) and is downregulated in human HCC and breast cancer (Tu and al., 2014).

lncRNA-17A BACE1, repressed GABABR2 variant A, promoted variant B, and enhanced accumulation of peptides Aβ42 and Aβ40 (Massone et al., 2011), BCYRN1 (Mus et al.,2007) and GOMAFU (MIAT)BC089918 GAS5 RMSTAS are associated with neurodegeneration. (Wapinski et al.; 2011Yu et al., 2013; Ng et al., 2013; Kim et al., 2013; Meier et al., 2010) and cancer (Mourtada-Maarabouni et al., 2009)

BACE1 protein levels and activity increase with brain aging and AD (Modarresi et al., 2011; Fukumoto et al., 2002)

7 SL interacts with the TP53 mRNA and suppresses p53 translation, during HuR can displace 7SL and increase p53 translation. lncRNA 7SLsuppresses TP53 (Abdelmohsen et al., 2014) and influences autophagy, senescence and cancer (Grammatikakis et al., 2014; Chen et al., 1997).

FOXP2 and mRNA in aging

But other RNAs are also an important component of aging, e.g., extracellular RNA play an important role in this process.(Douglas et al. 2016) 24–31nts piRNAs are responsible for proper genome in germ-line cells. 142.22 nts microRNAs cooperate with the 3′ UTR of target mRNAs binding RISC. This complex leads to mRNA degradation and disturbs protein translation. This affects components of the RNA spliceosome, 100–300 nucleotides Small nuclear RNAs (snRNAs), which process mRNAs in the nucleus. (Carthew et al. 2009) Aging relevant oxidative stress effects 5′ tRNA and 3′ tRNA fragments (Fu et al., 2009), which are necessary for cellular proliferation and interact with Argonauts proteins like extracellular miRNAs. (O'Brien et al., 2018) AGO2 is known to influence tumorigenesis through miRNAs-dependent or independent ways. (ZhenLong et al., 2015) Inflammatory miR-223 regulate the help of HDL ICAM-1 46 protein expression and snoRNAs, responsible for chromatin remodelling and rRNA modification. (Vickers and Remaley, 2017; Tabet et al., 2014) 200 nts non-coding lncRNAs regulate gene expression via chromatin association, Y-RNA 80–112 nts, coded in 4 genes, initiate DNA replication (Kheir & Krude, 2017; Yeri at al.2017) and are important for apoptosis. (Chakrabortty et al. 2015; Fritz et al., 2016). Circular RNAs, created from spliced exons and introns, influence tissue-specific development via miRNA. (Chen et al., 2015 Salzman et al., 2016). In endothelial cells miRNAs influence senescence, inflammation, cell differentiation and angiogenesis with the help of HDL Cockerill et al., 1995; Tabet et al., 1995, 2009; Schroen et al, 2012; Sun X, et al.,2012; Kane et al., 2012; Sumi et al. 2007;Pu and Liu L., 2008) Mi 223 also corelates with Hepatitis C healing. (Hyrina et al., 2017)

73 miRNAs e.g. miR-24-3p, miR-371a-5p, miR-3175, miR-3162-5p, miR-671-5p, miR-4667-5p, 146a-5p, 342-5p and miRs 5107-5p, which influence Gpx3 and thyroids, have significantly increased level in serum of old mouse. Level of 47 miRNAs e.g. miR-195a-5p and miR-34c-5p, also decrease with age. (Machida et al., 2015) This process can be influenced by caloric restriction. Mi RNAs also effects axon growth, melanogenesis, MARK, adherent and gap junctions (Victoria et al. 2015) and neuronal activity is affected by miRs 5107-5p, 146a-5p, and 342-5p. miR-5107-5 protects thyroids via Gpx3 and miR-195a-5p affect the aging relevant PI3K-AKT signalling pathway. miRNAs can also influence HTLV-1 infection, pluripotent stem cells, bone microarchitecture, osteoarthritis and cancer. (Hafez et al, 2015; Rodriguez-Fontenla et al. 2014; Wu et al., 2014) as well es aging relevant Bmpr1a,Vegfa, Bcl-2, Map2k1 Wnt4. MiR-34-5p influences GABAergic neurons, vascular aging response and cancer relevant Notch1, Notch4, Ccne2, E2f3, Gabra3 and Cylcin E2 through a Notch-dependent mechanism and Sirt1 (Li et al. 2011) Prop1 leads to decreased level of GH, Igf-1 and TSH. (Bartkeet al. 2001; Victoria et al., 2015) Lysine-, Valine-, Arginine-, Cysteine-, Glycine tRNA were decreased and Histidine- and Aspartic- acid derived 5′ tRNA increased in age. These changes were also CR sensitive (Dhahbi et al. 2013)

Aging is also associated with increased mRNAs level of CSF2, which is necessary for monocytes activation (Croxfordet al. 2015), granulocyte and macrophage maturation (Hamilton, 2002), energy homeostasis and metabolism. mRNA of subunit of pyruvate dehydrogenase enzyme DLD, which is necessary for exocytosis is also increased. RAB3 GTPase activating Protein Subunit 2 (Bem et al., 2011), an alternative splicing nuclear protein, which is involved in metastasis suppression with the help of splicing factor SRSF1 RRP1B (Lee et al. 2014), CSF2RA, SLC35B4, LAMB2 (Schaeffer, 2012), LAMB2 and LC35B4 showed increased mRNA level. Some lncRNAs were also hyperexpessed in ageing. At the same time 12 snoRNAs and 20 piRNAs were decreased. Decreased rRNA, which interacts with TTF1, may play a role in age related Alzheimer disease. (Freedman et al.,2016) miR-31 decrease is associated with aging relevant chronic myeloid leukaemia and ovarian carcinoma (Mitamura et al., 2013; Rokah et al.,2012; Korner et al., 2013; Zhong et al., 2013) miR-485-5p expression also alternates in aging. (Faghihi et al.,2008; 2010;Lee et al. 2014)

FOXP2 and mi RNA in oncological processes

Like the other genes in the FOXP family, the Fox P2 gene can be inactivated in cancer cells through miRNAs and alter its action this way. The recent study by Herrero and Gitton (2018) showed that FOXP2 can be blocked by TWIST activated miRs 199a-214, miR-762, miR-1915, let-7b and miR-34a and miR-3666. let-7a-d, miRs, miR-26a, miR-10,1miR-200b (which leads to increased MT1-MMPu und decreased PTEN- expression) (Soubani et al., 2012) and decreases prostaglandin, NF-kappaB,PEG2, VEGF activity ( Ali et al., 2010; Bao et al., 2011), EpCAM and EZH2 level and inhibits NOTCH-1 in pancreatic cancer (Bao et al., 2012). Wen-Zhuo et al. (2016) investigated the regulation of the FOXP2 gene by the microRNA-190 in gastric cancer. Valencia-Sanchez et al. (2006) showed that miRNAs destroy FOXP2-mRNA through its interaction with 3 'UTR, prevent its translation and this way negatively regulate the FOXP2 target genes. Many other studies also indicated that miRNA dysregulation may play an important role in the initiation and progression of oncological processes. (Barbrotto et al., 2008; Kasinski and Slack, 2011) Two years earlier the group of Wen-Zhuo et al. used the dual luciferase enzyme assays and confirmed that the miR-190 interacts with the FOXP2. RT-PCR and Western blot verified that miR-190 overexpression suppresses expression of FOXP2. Decrease of miRNA-190 expression leads in turn to FOXP2-mRNA and -protein increase. Because FoxP2 plays an important role in many oncological processes, the miR-190 could serve as a potential gastric cancer marker. Zhang et al. (2009) investigated various miRNAs and identified eight of them, which levels were increased in pancreatic cancer, e.g. the miR-190. This miRNA correlated with the progression of gliomas (Almog et al., 2012; Cuiffo et al.,2014) demonstrated suppression of FOXP2 expression by miR-199a in breast cancer mesenchymal cells. In lung, breast, bladder, colerectal and liver cancers MiRNA expression was also increased. (Ichimi et al., 2009; Lowery et al., 2009; Navon et al., 2009; Ura et al., 2009; Ng et al., 2009)

The results indicate that miRNAs play a different role in different tumor types at different stages. The influence of various miRNAs on pancreatic cancer (Zhang et al., 2009), lymphoma (Musilova and Mraz, 2014; Jones et al., 2014) and breast cancer (Wu et al., 2009) was also shown. It would be useful to study whether this miRNAs dysregulation also influences these and other cancers in connection with FOXP2?

Regulation of other FOXP genes by various factors on the example of carcinogenic processes In oncological tissue FoxP genes as well as other FOX genes can be modulated by microRNA . (Zheng et al., 2015), (Shao et al., 2013), (Kong et al., 2013), (Zhang et al., 2015), (Kundu et al., 2016), (Mei et al., 2015), (Yu et al., 2017). This modulation determines whether FOXP2 acts as an oncogene or as a tumor suppressor gene. In these processes the FoxP genes can also be deactivated by micro RNAs. (Choi et, 2016) According to Chang et al. (2007), He et al. (2010) and Raver-Shapira et al. (2007) the transcription factor p53 induces the transcription of the microRNA miR-34s, which in turn suppresses anti-apoptotic genes, which are required for cell development and growth. According to Choi et al. (2016), Isken et al. (2008), Mraz et al. (2009), Zenz et al. (2009), miR-34a plays an important role in both chronic lymphatic and acute myeloid leukaemia and the Foxp1 is regulated by miR-34a. The miR-34a also builds a direct link between the tumor suppressor p53 and the oncoprotein Bcl-2. Rao et al. (2010) pointed out that in B lymphocytes too miR-34 modulates Foxp1 via the tumor suppressor gene p53. The tumor suppressor p53 could also be a direct FOXP2 target gene. (Herrero and Gitton, 2018)There are also further indications of androgen influences on the FoxP1 expression and associated carcinogenic processes. (Banham et al, 2007; Bates et al., 2008; Fox et al., 2004; Giatromanolaki et al., 2006; Takayama et al., 2008) Further FOXP1 appears to affect follicular lymphoma as well as in aging dependent ovarian and colorectal carcinoma. (Brown et al., 2004; Choi et al., 2016; De Smedt et al., 2015) In B cell lymphomas or hepatocellular carcinomas FOXP-1 serves as an oncogene. (Zhang et al., 2012; Jiang et al., 2012; Xia et al., 2014) In lungs, prostate and endometrial tumors FOXP1 is a potential predictor of disease progression. (Feng et al., 2012; Toma et al., 2011; Giatromanolaki et al., 2006; Takayama et al., 2008; Takayama et al., 2014)

FOXP2 and neuroplasticity

FOXP2 plays an extremely important role in neuroplasticity (Haesler et al., 2006; Ebisu et al., 2017; Konopka et al., 2009; Boeckx et al., 2014; Dodson et al., 2015; Garcia-Calero et al., 2016) and review of Wohlgemuth et al. (2014) showed that FOXP2 also provided the NMDAR-mediated neuronal plasticity affecting MAPKK and tyrosine phosphatase. The MARK is a RAS downstream effector. RAS is a a GTPase that indirectly interacts with oncological relevant ant p21 and p53 indirectly. RAS, in turn, is activated by growth factor signalling and interacts with FOXO-inhibiting PIK3. Further interaction with neuro and oncoactive Trk is also discussed. (Santos, 2011), (Mengeselt et al., 2008) It influences cell proliferation and this plays a role in the development of cancer. A direct FOXO4-p53 interaction and its importance in cell aging has been demonstrated too. (Baar et al., 2017) It would be interesting to look more closely at the role of FOXP in these processes. Possibly Fisetin, related to Qis (Zhu et al., 2017), but also FOXO4-related peptide that inhibits the PI3K/AKT/p53/p21/Serpine SCAP (Zhu et al., 2015b), recently noted to be senolytic (Baar et al., 2017) could have positive effect on cancer and aging.

Guibinga et al. showed in "Neuropathogenesis of LNS. Striatal Neurodevelopment is Dysregulated in Purine Metabolism Deficiency and Impacts DARPP-32, BDNF / TrkB Expression and Signaling: New Insights on the Molecular and Cellular Basis of Lesch-Nyhan Syndrome" (2014) that downregulating Bcl11b in HPRT-deficient cells elicited a change in DARPP-32 level in the neural stem cells. This phosphoprotein is regulated by dopamine and cAMP. Its altered level in turn led to striatal B-cell leukaemia 11b (Bcl11b). The activator of the DARPP-32 striatal BDNF / TrkB signalling pathway was highly increased in HPRT-deficient cells as well as in the striatum of the HPRT knockout mice. This protected the cells from reactive oxygen species (ROS)-mediated cell death.

There are also FOXP2 interactions with members of the histone family (H2AFX; H3f3B), two heat shock proteins (Hsp25; Hsp90a) and Calcycline-binding proteins ARHGEF9, CtBP1 and effectors Gli1, Gli2 and Gli3, EGR1, IEGs, D1R and DARPP32. In addition, FOXP2 affects NMDAR-mediated neuronal plasticity, MAPKK and tyrosine phosphatase. (Mc Auley et al., 2017)

γH2AX foci in aged HSCs is increased (Rossi et al., 2007; Beerman et al., 2014; Moehrle et al., 2015; Flach et al., 2014)

FOXP2 and neurodegeneration

Several studies showed the significance of FoxP2 for the brain and skull development (Benítez-Burraco et al., 2015) but above all for the language. (Watkins et al., 2002; Vargha-Khadem et al., 1998; Middleton ad Strick, 2000; Watkins et al., 2002; Liegeois et al., 2003; Lai et al., 2003). The FOXP2 gene (formerly also known as SPCH1, TNRC10 or CAGH44) was mainly associated with important language functions. First, the FOXP2 became known through the work of Anthony Monaco and Svante Pääbo Group at the Institute for Evolutionary Anthropology of the Max Planck Society in Leipzig. They studied the linguistic deficient KE family and found an autosomal dominant missense point mutation in exon 14 of the 7th chromosome (in the 7q31 band guanine was replaced by adenine). This mutation replaced arginine (R) by histidine (H) at the position 553 R553H of the FOXP2 protein and caused an inhibition of the DNA-binding domain and the inoperability of the protein. Other FOXP2 mutations were also observed by dyspraxia patients. (MacDermot et al., 2005; O'Brien et al., 2003; Jiménez-Romero et al., 2016; Becker et al., 2015; Zeesman et al., 2006).

In 2016 described Tborres-Ruiz et al. a girl with inherited complete chromosomal rearrangement. This rearrangement was accompanied by a fracture proximal and distal to the FOXP2 gene and resulted in cognitive disabilities. The fracture in a new neuroblastoma cell line SK-N-MC led to the decline of FOXP2 and increase of MDFIC-P protein levels. The MDFIC-P gene is localized near FOXP2. The FOXP-2 significance for the childhood apraxia of speech (CAS) and other speech disorders were also described. (Morgan et al., 2017; Kurt et al, 2012)

The FOXP2 mutations are often accompanied by structural brain changes, so the new multimodal MRI study of an eight-year-old boy (A-II) with a de novo FOXP2 deletion by Liégeois et al. „Early neuroimaging markers of FOXP2 intragenic deletion“ (2016). The researchers described in their work significant bilateral structural abnormalities in the basal ganglia and in hippocampus. In the hippocampus, in the thalamus, in the globus pallidum and in the caudate nucleus a volume reduction was also observed in comparison to the control group of 26 healthy volunteers. The patients showed no detectable functional MRI activity by the repetition of nonsense phrases.

FOXP2 also plays a role in the visual system (Iwai et al., 2013; Horng et al., 2009), in psychiatric disorders, and in aging dependent frontotemporal degenerative dementia accompanied by speech disorders (Sanjuán et al., 2006; Park et al., 2014; Wang et al, 2016; Bacon and Rappold, 2012; Fisher and Scharff, 2009; Premi et al., 2012; Kumar, et al., 2011).

FOXP2 is important for brain development and communication in many animal taxa (e.g. songbirds, marine mammals, bats and possibly elephants) which learn to communicate through imitation and whose auditory processing needs to interact with motor control. (Scharff and Haesler, 2005)

Scharff and Petri (2011) emphasized in „Evo-devo, deep homology and FoxP2: implications for the evolution of speech and language“ that FOXP2 shows different expression cycles in different neuronal subtypes and „the more efforts should be directed at identifying the genomic loci regulating temporal expression differences of FoxP2“.

Other members of the FOXP2 family show many parallels to FOXP2. (Viscard et al., 2017)

FOXP2 and Alzheimer

López-González et al., 2016 described in FOXP2 Expression in Frontotemporal Lobar Degeneration-Tau that reduced mRNA and protein expression of FOXP2 in frontal cortex area 8 in Pick’s disease and in frontotemporal lobar degeneration-tau. This tau degeneration was linked to P301L mutation, that was associated with language impairment in comparison to age-matched controls and cases with parkinsonian variant progressive supranuclear palsy. “Foxp2 mRNA and protein are also reduced with disease progression in the somatosensory cortex in transgenic mice bearing the P301S mutation in MAPT when compared with wild-type littermates. These findings support the presence of FOXP2 expression abnormalities in sporadic and familial frontotemporal degeneration tauopathies.” (López-González et al., 2016, p.1)

Padovani et al. investigated 2010 in „The speech and language FOXP2 gene modulates the phenotype of frontotemporal lobar degeneration“ the influence of genetic variations within FOXP2 in neurological disorders and how FOXP2 polymorphisms influences frontotemporal lobar degeneration. After neuropsychological examination as well as brain imaging of two-hundred ten FTLD patients and in 200 age-matched healthy controls the researchers evaluated four FOXP2 polymorphisms: rs2396753, rs1456031, rs17137124 and rs1852469 they observed no significant differences in SPECT images of the four FOXP2 polymorphisms in genotype distribution and allele frequency between FTLD and controls, in the same time they reported a significant and specific association between rs1456031 TT and rs17137124 TT genotypes and verbal fluency scores and an addictive effect of two polymorphisms. Afterwards they computed the number of observations over time and obtained 391 comparable results that showed: FTLD patients carrying at-risk polymorphisms have greater hypoperfusion in the frontal areas, namely the left inferior frontal gyrus, and putamen, compared to the non-carriers. Genetic variations within FOXP2 modulate FTLD presentation when disease is overt, affecting language performances and leading to hypoperfusion in language-associated brain areas.

Di Meco et al. proposed 2019 in „Gestational high fat diet protects 3xTg offspring from memory impairments, synaptic dysfunction, and brain pathology“ new insights in maternal history for sporadic Alzheimer’s disease (AD) and the possibility of susceptibility modulation to AD via gestational high fat diet. Triple transgenic dams (human PS1, human MAPT and KM670/671NL)

were fed with high fat diet: 42% calories from fat or regular chow 13% calories from fat throughout 3 weeks gestation. This study showed how gestational high fat diet attenuated memory decline, synaptic dysfunction, amyloid-β and tau neuropathology (decrease in the levels of Aβ1–40 and Aβ1–42) in the offspring by transcriptional regulation of BACE-1, CDK5, and tau gene expression via the upregulation of FOXP2 repressor. To proof that FOXP2 effects on tau,

CDK5 and BACE-1 the researchers exposed APPswe mutant N2A cells to FOXP2-GFP plasmid and measured tau, FOXP2, CDK5 and BACE-1 protein and mRNA levels 24 hours after, wich were decreased, but lower APP level was not observed. Behavioral impairments could be improved under and electrophysiology of 3TG hippocampal slices showed higher excitatory postsynaptic potentials as well as increasing strength of stimulus intensities. Longterm potentiation in the CA1 region of the hippocampus and paired pulse facilitation were measured and showed partial restoration of the

fEPSP in 3TG HF, also of neuronal plasticity and memory. So gestational high fat diet significantly decreases tau aggregation-prone isoforms level, soluble tau level as well as of its phosphorylated isoforms and protects this way offspring from later AD.

There is many evidences about indirect influence of FOXP2 on neurodegenerative diseases, so:

FOXP2 controls expression of for Alzheimer's disease relevant RELN (Adam et al.,2016)

Seripa et al. described 2012 in „The RELN locus in Alzheimer's disease. “ serine protease, encoded by the RELN gene, as part of in AD involved apoE pathways. The researchers investigated three polymorphisms in the RELN locus, i.e., a triplet tandem repeat in the 5'UTR and two single-nucleotide polymorphisms (SNPs) rs607755 and rs2229874, located in the exon 6 splice-junction and in the exon 50 coding region. The analysis of 223 sporadic AD patients 181 controls. There was no significant differences in rs2229874 but in 5'UTR and rs607755 genotypes, in females and not in males (even if APOE genotypes were adjustment). So RELN gene variants may effect AD pathogenesis of, especially in females.

In“ Reelin depletion is an early phenomenon of Alzheimer's pathology“ 2012 Herring et al. examined the expression profile of with Alzheimer's disease associated “RELN and its downstream signalling members APOER2, VLDLR, and DAB1 in AD-vulnerable regions of transgenic and wildtype mice as well as in AD patients and controls across disease stages and/or aging..”(Herring et al., 2012, p.1) Their results showed that “AD pathology and aging are associated with perturbation of the RELN pathway in a species-, region-, and molecule-specific manner” and the “depletion of RELN, but not its downstream signalling molecules, is detectable long before the onset of amyloid-β pathology in the murine hippocampus and in a pre-clinical AD stage in the human frontal cortex. This early event hints at a possible causative role of RELN decline in the precipitation of AD pathology and supports RELN's potential as a pre-clinical marker for AD.” (Herring et al., 2012, p.1)

FOXP-2 affects NCAM1, VLDLR and other target genes in the nervous system

FOXP2 controls expression of for Alzheimer's disease relevant glycoprotein NCAM1. (Atz et al., 2007) ( Konopka et al., 2009) The FOXP2 also influences DISC1, (Walker et al., 2012), (Miyoshi et al., 2004), a Reelin receptor VLDLR (Adam et al., 2016), NURR1, PHOX2B, TBX22, SEBOX and FOXL1, CDH4 and CDH11, DICER1, RISC TARBP2, DICER1, EIF2C1-4 , DCDC2, KIF13B, PTPRQ, MSN, FOXP2 , NAKR1, SEBOX, MARVELD1, PHOX2B, MYH8, MYH13, PIK3K, PTPRQ, PIM1, NFE2L2, ERP44, KEAP1, JAK / STAT signalling, the phosphatase PTEN, BACE2, SERPINH1, CDH4 and the Ezrin-Radixin-Moesin complex. These proteins are involved in nervous system myelination, neuroinflammation, amyloid precursor protein formation, Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Lewy body dementia and Parkinson's disease (Devanna et al.,2014) Different of these targets play an important role in aging and can be affected via caloric restriction. The amount of satellite cells decreases with age (Brack et al., 2005; Collins et al., 2007; Gibson and Schultz, 1983) and like hematopoietic stem cells they change their Wnt and Notch pathways with the age as well as TGF-β and sFGF ligands and differentiate less to myogenic lineage and more to fibrogenic lineage Brack et al., 2007 Carlson et al., 2009 Carlson and Faulkner, 1989 Chakkalakal et al., 2012 ; Conboy et al., 2003, 2005; Sinha et al., 2014) and cytokine signalling via the JAK-STAT pathway (Price et al., 2014) and increase p38-MAPK signalling (Bernet et al., 2014; Cosgrove et al., 2014) Sousa-Victor et al., 2014 ). Wnt3, GH, TGF-β and IGF improve neurogenesis (Blackmore et al., 2009 Katsimpardi et al., 2014 ; Lichtenwalner et al., 2001; Okamoto et al., 2011; Pineda et al., 2013 ; Villeda et al., 2014 ).Growth differentiation factor 11 (GDF11)can improve NSC- and satellite cell function, but ist production decreases with aging. (Katsimpardi et al., 2014; Loffredo et al., 2013.; Sinha et al., 2014). At the same time high TGF-β levels disturb satellite cells and neuronal stem cells function (Carlson et al., 2009;) but growth differentiation factor 11 improves it. (Katsimpardi et al., 2014 ; Loffredo et al., 2013; Sinha et al., 2014).

Kanekiyo and Bu showed 2014 that Low-density lipoprotein receptor-related protein 1 regulates cellular Aβ uptake and degradation in neurons, astrocytes, and microglia in brain parenchyma, and in vascular smooth muscle cells and pericytes in cerebrovascular. It also mediates Aβ clearance at the BBB by facilitating Aβ transport from brain to blood and Apolipoprotein E is a major ligand for LRP1 and influences AD risk by affecting Aβ aggregation, cellular uptake and degradation, apoE and Aβ “can interact with each other, they also share common receptors including LRP1, LDLR, and HSPG on cell surface. ApoE likely competes with Aβ for their receptor binding but can also facilitate cellular Aβ uptake by forming apoE/Aβ complexes depending on their concentrations, apoE isoform involved, lipidation status, Aβ aggregation status and receptor distribution patterns. Dissecting how LRP1 participates in apoE-mediated Aβ clearance will be critical to develop apoE-targeted therapy for AD.” (Kanekiyo and Bu, 2014, p. 7)

FOXP2 controls expression of for Alzheimer's disease relevant PTEN (Oswald et al.,2017; Frere and Slutsky, 2016; Knafo et al., 2016; Zhang et al. 2006; Rickle et al., 2006) Mislocalization of Pten in murine brain was observed to correlate with down-regulation of Foxp2 and upregulation of Msn (Tilot et al., 2016)

Further FOXP2 controls expression of for Alzheimer's disease relevant glycoprotein NURR1 Nurr1 was specifically expressed in glutamatergic neurons of the hippocampus of healthy brains and that these Nurr1‐expressing, Aβ‐positive glutamatergic neurons degenerated in an age‐dependent manner in 5XFAD mice and plays important roles in AD pathogenesis. (Moon et al.Nurr1, 2019)

According to Oswald et al. (2017) „The FOXP2-Driven Network in Developmental Disorders and Neurodegeneration“ the transcription factor encoded by the new FOXP2 target NURR1 (also NR4A2, NOT) seems to be of special importance for normal dopaminergic functioning. So stimulation of NURR1 improves behavioural deficits, associated with the degeneration of dopamine neurons in PD model mice – an effect which involves enhanced trans-repression of neurotoxic pro-inflammatory genes in microglia and increased transcriptional activation of midbrain dopaminergic (mDA)neurons (Kim et al., 2015). Nurr1 knockout mice even fail to develop dopamine neurons (Zetterström et al., 1997).

So dopamine-related diseases AD, PD SCZD, Lewy body dementia are accompanied by several NURR1 mutations. (e.g., Chen et al., 2001; Zheng et al., 2003; Chu et al., 2006).

FOXP2 controls expression of for Alzheimer's disease relevant glycoprotein NCAM1. (Gillian et al., 1994; Todaro et al.,2004 )

Moreover, NCAM1 is a putative target of both RUNX2 (Kuhlwilm et al., 2013) and FOXP2

(Konopka et al., 2009). In Boeckx and Benítez-Burraco “Globularity and language-readiness: generating new predictions by expanding the set of genes of interest”, the FOXP2 also influences Alzheimer relevant DISC1. Using dual luciferase assays Walker et al. (2012) demonstrated that a region -300 to -177 bp relative to the transcription start site (TSS) contributes positively to DISC1 promoter activity, while a region -982 to -301 bp relative to the TSS confers a repressive effect and inhibition of DISC1 promoter activity and protein expression by forkhead-box P2 (FOXP2). R553H and R328X FOXP2 point mutations found, known from developmental verbal dyspraxia affected families, decreases his inhibition. Further knockdown of DISC1 increases the expression of APP at the cell surface and decreases its internalization. (Shahani et al., 2015)

Further aging and neurodegeneration relevant FOXP2 targets

FOXP2 and Vitamin D effect on ZNS

FOXP2 target RUNX2 binds to the 1alpha, 25-dihydroxyvitamin D3 receptor. (Paredes et al., 2004). The 25-dihydroxyvitamin D3 receptor is encoded by VDR involved in immune response and tumor suppression. Together with the VDR express multiple immune relevant genes, eg. they regulate SPAG5. (Stephens and Morrison., 2014). The SPAG5 encodes a protein required for the correct function of mitotic spindles, which also regulates cell stress-induced apoptosis. (Thedieck et al., 2013) Together with FOXP2, RUNX2 regulates aging relevant vitamin D (Patrick and Ames, 2014), (Hawes et al., 2015) and other interaction partners, such as: CREB (Oury et al., 2010)

Aging relevant H3K27ac and H3K14ac are acetylated via p300/CBP and its co-activator CREB . cAMP responsive CREB expression is responsible to fasting. So CBP, CREB, CRTC2 and TAF-4 activate together gluconeogenesis genes (Altarejos and Montminy, 2011 )

The RUNX2 also affects GTF2I (Lazebnik et al., 2009), there is some evidence for feedback processes because both GTF2I and RUNX2 expression are regulated by the AUTS2. (Oksenberg et al., 2014) The AUTS2 cooperates with the PRC1, the GTF2I, the SATB2, the ZMAT3, the RELN and the TBR1.

Vitamin D-Esr1-Igf1 interaction effects molecular pathways relevant to Alzheimer’s disease and Molecular Neurodegeneration. (Landel et al.,2016)

Kaneko and colleges demonstrated that calcitriol regulates the expression of two human brain-related genes containing VDREs, tryptophan hydroxylase (Tph) and leptin. Landel et al, 2016 studied the effect of maternal vitamin D deficiency on fetal brain development and identified that these genes are also modulated in the brains of either Wt or Tg mice and found that the pups from deficient mothers display a modulated expression of Bdnf, Foxp2, Tgfb1 and Th, which are also affected in certain conditions of this study. Together with FOXP2, RUNX2 regulates aging relevant vitamin D (Patrick and Ames, 2014), (Hawes et al., 2015) and other interaction partners, such as: CREB (Oury et al., 2010).

Vitamins and aging

In general vitamins are also age-related factors. So, Vitamin D is important for ROS protection in the ZNS and for cell cycle regulation.( Pusceddu, 2015) Vitamin B2 is an antioxidant due to its involvement in the glutathione redox cycle (in glutathione reductase (Ashoori, 2014) and it is a cofactor in amino acid and lipid metabolism as well as in redox reactions. Riboflavin reduction increases lipid peroxidation. (Wang, 2011) Vitamin B6 reduces homocysteine concentrations and protects against cardiovascular diseases (Okura , 2014) Vitamin B12 is a cofactor for the methionine synthase (important for folate cycle and homocysteine re-methylation) and for the Methylmalonyl CoA mutase ( Hughes, 2013) Aging is associated with B12 reduction. Antioxidant Vitamin C (ascorbic acid) and dehydroascorbic acid are necessary for myelin, peptide amination, for synthesis of neurotransmitters and carnitine and it helps to recycle vitamin E and tetrahydrobiopterin, its deficiency is associated with amyloid-β plaques. ( Dixit, 2015) Like vitamin E (Gutierrez-Fernandez, 2015; Rinaldi, 2003) Vitamin A and retinoic acid are necessary for neurodevelopment (Touyarot , 2013) and its reduction is associated with aging, inflammation. It influences p21 and Alzheimer disease.

FOXP2 influence on RA receptor expression and its effect on the retinoic acid-mediated neuronal differentiation

Benítez-Burraco and Boeckx (2014) described in „FOXP2 drives neuronal differentiation by interacting with retinoic acid signaling pathways“ the importance of RA signalling and of FOXP2 for brain processes, the upregulation of RARβ by FOXP2 . The FOXP2 indirectly regulates the SIRT1 and other genes via RUNX-UTS2-TBR1-DYRK1A cascade and directly some SIRT1 target genes. The DYRK1A in turn phosphorylates the SIRT1. So there exists possible a connection between the FOXP2 and the RUNX2 via SIRT1. In addition, the SIRT1 directly controls thewith nuclear retinoid receptor proteins termed as retinoic acid receptors (RARs) and retinoid X RUNX2.The Dyrk1A also promotes de-acetylation of TP53, which is associated with carcinogenesis (Ni et al., 2005). TP53 induces PANDA lncRNA, which influence aging via binding the transcription factor NF-YA.

Sodhi and Singh, 2014 found in „Retinoids as potential targets for Alzheimer's disease“ that vitamin A and its derivatives, the retinoids, modulate several physiological and pathological processes through their interactions with nuclear retinoid receptor proteins termed as retinoic acid receptors (RARs) and retinoid X receptors (RXRs). Both have an antioxidant potential. Retinoid signalling exists in including amygdala, hypothalamus, hippocampus, striatum and cortex and its defects seem to be relevant for defects in learning, memory and for Alzheimer's disease. Retinoids also decrease pro-inflammatory cytokines- and chemokines-level by astrocytes and the microglia. RA exposure leads to an up-regulation of choline acetyltransferase (ChAT) level and activity, ameliorated the symptoms of AD and reduced amyloid accumulation and tau hyperphosphorylation in APP/PS1 transgenic mice and its isomers enhance, the expression of genes linked with cholesterol efflux e.g. apoe, abca-1 and abcg-1 proteins in astrocytes.

Also according to Das et al..2019 „Potential therapeutic roles of retinoids for prevention of neuroinflammation and neurodegeneration in Alzheimer's disease“ retinoids have an important impact on neural patterning, differentiation, axon outgrowth and brain function, impaired RA- signalling leads to oxidative stress, mitochondrial malfunction, neuroinflammation, neurodegeneration and Alzheimer's disease (associated with aggregated amyloid-beta and hyperphosphorylated tau protein). They also described loss of spatial learning and memory as a result of low RA-level, because retinoids inhibit expression of chemokines and neuroinflammatory cytokines in microglia and astrocytes, which are activated in Alzheimer's disease. Retinoic acid receptors stimulation decreases amyloids accumulation,neurodegeneration level, and thereby prevents pathogenesis of Alzheimer's disease in mice.

Shudo et al. published 2009 „Towards retinoid therapy for Alzheimer's disease. “This paper deals mainly with AD in relation to retinoic acid receptors (RARs: RARα, β and γ) and their ligands (retinoids), such as the endogenous RAR ligand all-trans-retinoic acid (RA), considering current knowledge about PD, ALS and other neurodegenerative diseases. “It is important to note that factors leading to the onset of these diseases are still poorly understood, and so there is a great deal of scope for novel therapeutic approaches. Recent findings indicate that the window of opportunity for enhancing or normalizing the growth of neuronal cells and promoting recovery from neurodegenerative diseases may be larger than previously thought.” (Shudo et al., 2009, p.1)

Another direct FOXP2 target is the general transcription factor GTF3C3, which plays an important role among others in apoptosis (Zhan Y, 2002).

According to Devanna et al. (2014) "FOXP2 drives neuronal differentiation by interacting with retinoic acid signalling pathways", FOXP2 interaction with retinoic acid makes cells more sensitive to RA effects and strengthens this way neuronal differentiation. This leads to increased neurite growth and branching as well as to decreased neuronal migration. These effects are particularly important in the striatum because speech-disabled people with a mutant FOXP2 gene have a pathology in this brain area. This gives further hint to the FOXP2 role in neuronal differentiation. The authors also mentioned that FOXP2 reduces DDL3 and RARβ (retinoic acid receptor) expression in the striatum.

In “Retinoic Acid Signaling: A New Piece in the Spoken Language Puzzle” (Rhijn et al. 2015) looked the researchers for evidence that the FOXP2 and RA pathways overlap. They analysed molecular, cellular and behavioural levels and found that FOXP2 changes RA receptor expression. These receptors directly control cellular response to RA. The retinoic acid receptor β (RAR β) was of particular interest because mice with the corresponding mutation showed severe movement deficits and its motor learning was severely impaired. (Krezel et al.) Increased RA level in pregnant rats led to behavioural problems and to impaired learning, memory and motor function. (Holson et al., 1997) Rats with a vitamin A deficiency also showed poor motor performance when they learned new movements. (Carta et al., 2006) In addition vitamin deficit had a negative impact on striatal development. Striatal progenitor cells could not differentiate without RA signals (Krezel et al., 1998), (Chatzi et al.,2011). Acute RA-level reduction in mice led to impaired induction in synaptic grading and impairment of hippocampal LTP and LTD, which was, however, reversible. Normal synaptic plasticity was quickly restored in this phenotype with the help of vitamin A supplementation. (Misner et al., 2001)

According to Boccardi et al. „Beta-carotene, telomerase activity and Alzheimer's disease in old age subjects“, 2019 β-carotene significantly and positively correlated with telomerase activity, independent of gender, Β-carotene plasma level was associated with AD diagnosis and

β-carotene may modulate telomerase activity in old age. Moreover, lower plasma β-carotene levels, correlating with peripheral telomerase activity, are associated with AD diagnosis independent of multiple covariates. This way FOXP2 genes can have a further effect on aging and tumorigenesis.

According to Devanna et al. (2014) „FOXP2 drives neuronal differentiation by interacting with retinoic acid signalling pathways“ FOXP2 reduces the expression of DDL3 and RARβ (retinoic acid receptor). The CARET study showed that high-dose beta-carotene (a precursor of retinoic acid (vitamin A) for an extended period was suspect to cause by 18 percent smokers lung cancer and it is known that FoxP2 is high expressed in lungs (Li, et al., 2004; Zhou et al., 2008;Groszer et al., 2008). FoxP2-coexpression with the transcription factor homeodomain Nkx2.1 in the lung was described by Li et (2012) FoxP2 interaction with the CtBP1 co-repressor may be involved in tumor suppression of breast cancer. CtBp interacts with the oncofactor BRCA1 / 2 (Chinnadurai G., 2009) (Deng et al. 2012) It would be of great interest to investigate whether the pathogenic vitamin A effect in this case is due to the interactions with FOXP2 which play a role in many oncological processes.

CDH4

According to Liu et al., 2012 total cerebral brain volume depends on CDH4-level, involved also in AD.These findings suggest that Dicer1 may be a target in AD therapy.

DICER1

Yan et al. (2019) explained that Dicer1 is reduced in APPswe/PSEN1dE9 mice and is regulated by Nrf2 and Brain Dicer1 is down-regulated in a mouse model of Alzheimer’s disease via Aβ42-induced repression of nuclear factor erythroid 2-related factor 2. The researchers studied unexploited roles of Dicer1 in AD and a novel way of Dicer1 regulation. Their results make hope that Dicer1 may be a target in AD therapy.

TARBP2

According to Haroon et al., 2016 „A designed recombinant fusion protein for targeted delivery of siRNA to the mouse brain“TARBP2 Binding Protein fused to a brain targeting peptide that binds to monosialoganglioside GM1. “Conformation-specific binding of TARBP2 domain to siRNA led to the formation of homogenous serum-stable complex with targeting potential. Further, uptake of the complex in Neuro-2a, IMR32 and HepG2 cells analysed by confocal microscopy and fluorescence activated cell sorting, revealed selective requirement of GM1 for entry. Remarkably, systemic delivery of the fluorescently labelled complex (TARBP-BTP:siRNA) in ΑβPP-PS1 mouse model of Alzheimer's disease (AD) led to distinctive localization in the cerebral hemisphere. Further, the delivery of siRNA mediated by TARBP-BTP led to significant knockdown of BACE1 in the brain, in both ΑβPP-PS1 mice and wild type C57BL/6. The study establishes the growing importance of fusion proteins in delivering therapeutic siRNA to brain tissues.” (Haroon et al., 2016, p. 1)

PIK3K

Gabbouj et al. (2019) describe in „Altered Insulin Signaling in Alzheimer's Disease Brain - Special Emphasis on PI3K-Akt Pathway“ the PI3K-Akt signalling pathway, involved in microglia and astrocytes, as an important player in T2D pathogenesis and insulin mediation. Decreased levels of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) subunits and decreased Akt kinase phosphorylation is associated with AD, amyloid-β and tau pathologies. TWD intake leads to altered PI3K subunits-levels and of intranasal insulin to enhancement of PI3K-Akt signalling, improved memory in human trials.

PIM1

PRAS40 phosphorylation is regulated by Pim1 Velazquez et al. (2016) gave a strong evidence for interconnection between the mammalian target of rapamycin (mTOR), proline-rich AKT substrate, PRAS40-phosphorylation-levels and Aβ, tau pathologies and cognitive deficits.

BACE1

Das, B. and Yan, R. (2017) described in „Role of BACE1 in Alzheimer's synaptic function“ that Aβ is generated from amyloid precursor protein (APP) via proteolytic cleavage by β-site APP cleaving enzyme 1 (BACE1) and BACE1 inhibition reduces Aβ-level in humans. BACE1 inhibitors could be an effective AD remedy.

NFE2L2

Otter et al. (2010) illustrated in „Nrf2-encoding NFE2L2 haplotypes influence disease progression but not risk in Alzheimer's disease and age-related cataract“ how one haplotype allele of NFE2L2 gene, encoding the main regulators of the defence system against oxidative stress, age-related cataract and AD, Nrf2-protein, was associated with 2 years earlier age at AD onset and 4 years earlier age at surgery for posterior subcapsular cataract.

According to Joshi and Johnson (2012)“The Nrf2-ARE pathway: a valuable therapeutic target for the treatment of neurodegenerative diseases“ neurodegenerative relevant NF-E2 related factor-overexpression has a positive impact on Amyotrophic lateral sclerosis, Alzheimer’s disease and Parkinson. A cis-acting antioxidant response element regulates phase II detoxification enzymes via ARE-Nrf2 binding with the help of Keap1, a culin 3-based E3 ligase that targets Nrf2 for degradation, sequesters Nrf2 in cytoplasm. Disruption of Keap1-Nrf2 interaction or genetic overexpression of Nrf2 has a positive effect on oxidative stress.

Pajares et al. (2016) identified in „Transcription factor NFE2L2/NRF2 is a regulator of macroautophagy genes“ the transcription factor NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2) as a regulator of autophagy gene expression and its relevance to amyloid β precursor protein, MAPT/TAU and AD. According to ENCODE for BACH1 and MAFK, that bind the NFE2L2-regulated enhancer ARE, 27 putative AREs in 16 autophagy-related genes were identified and twelve of these sequences were validated as NFE2L2 regulated AREs in 9 autophagy genes after NFE2L2 activation with sulforaphane.

Saad El-Din et al. (2020) describe in „Active form of vitamin D analogue mitigates neurodegenerative changes in Alzheimer's disease in rats by targeting Keap1/Nrf2 and MAPK-38p/ERK signaling pathways“ the Nrf2 as a promising target for the prevention of Alzheimer's disease and vitamin D, its analogue, Maxacalcitol as crucial for improving AD cognitive functions via Keap1-Nrf2 signalling pathway.

Rojo et al. (2017) also confirmed in „NRF2 deficiency replicates transcriptomic changes in Alzheimer's patients and worsens APP and TAU pathology“ NRF2 as a crucial regulator of multiple stress responses, which also protects against inflammation and proteotoxicity and ageing is associated with decline of its level. Young adult AT-NRF2-KO mice showed deficits in long term potentiation in the perforant pathway, learning and memory.

Bahn et al. (2019) showed in „NRF2/ARE pathway negatively regulates BACE1 expression and ameliorates cognitive deficits in mouse Alzheimer's models“ BACE1 as the rate limiting Aβ generation enzyme. AD is accompanied by BACE1 and a BACE1 mRNA-stabilizing antisense RNA elevation. NRF2/NFE2L2 represses the BACE1 and BACE1-AS-expression via ARE promoters binding, independent of redox regulation. Also NRF2 improves cognitive deficits in animal models of AD, so the authors regard NRF2 as a possible key factor in prevention of early pathogenic process in AD.

KEAP1

Kerr et al. (2017) associate in „Direct Keap1-Nrf2 disruption as a potential therapeutic target for Alzheimer's disease“ Nrf2 with cell protection and an attractive therapeutic target for the prevention of neurodegenerative diseases, including Alzheimer’s disease (AD), provided in vivo evidence that specific inhibition of negative regulator of Nrf2 Keap1 can prevent neuronal toxicity in response to the AD-initiating Aβ42 peptide. Lithium, an inhibitor of the Nrf2 suppressor GSK-3, prevented Aβ42 toxicity in Nrf2 independent way.

JAK / STAT signalling

Nevado-Holgado et al. published 2019 „Genetic and Real-World Clinical Data, Combined with Empirical Validation, Nominate Jak-Stat Signaling as a Target for Alzheimer's Disease Therapeutic Developmen“, where they combined GWAS results with the current knowledge of molecular pathways, real-world clinical data from six million patients, RNA expression across tissues from AD patients and rodent models and showed that the degree of comorbidity of these diseases with AD correlates with the strength of their genetic association with molecular participants in the Janus kinases/signal transducer and activator of transcription pathway. They demonstrated Aβ induction by JAK-STAT anomalies and identified these genes as a potential target for therapeutic approach.

SERPINH1

Aβ and cytokines, involved in microglial activation, play a crucial role in neuroinflammation and AD. Yoo et al. published 2015 „ Amyloid-beta-activated human microglial cells through ER-resident proteins“ . They performed a proteomic analysis of Aβ-stimulated human microglial cells by stable isotope labelling with amino acids in cell culture combined with LC-MS/MS and clarified ER-resident proteins-level of PDIA6, PDIA3, PPIB and SERPINH1 was altered by 1.5 fold or greater. The researchers suggested that ER proteins play an essential role in human microglial activation by Aβ and could be important therapeutic targets for treatment of AD.

Ezrin-Radixin-Moesin complex.

α-secretases cleave the amyloid precursor protein to neuroprotective soluble APP ectodomain. Darmellah et al. (2012) show in „ Ezrin/radixin/moesin are required for the purinergic P2X7 receptor (P2X7R)-dependent processing of the amyloid precursor protein“ that the activation of ezrin, radixin, and moesin proteins is required for the P2X7R-dependent proteolytic processing of APP leading to sAPPα release and the ERM down-regulation via siRNA blocked it and P2X7R stimulation triggered its phosphorylation. Ezrin must translocate to the plasma membrane to interact with P2X7R and enzymes Rho kinase and the MAPK modules ERK1/2 and JNK act upstream of ERM, whereas a PI3K activity is triggered downstream.

Vega et al. (2018) also demonstrated in „Ezrin Expression is Increased During Disease Progression in a Tauopathy Mouse Model and Alzheimer's Disease“ that the increased Ezrin-level leads to the early stages of neurodegeneration in tauopathy models and human disease.

According to Oswald et al. (2017) „The FOXP2-Driven Network in Developmental Disorders and Neurodegeneration“ these proteins are involved in nervous system myelination, neuroinflammation, amyloid precursor protein formation, Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Lewy body dementia and Parkinson's disease (Devanna et al.,2014) Different of these targets play an important role in aging and can be affected via caloric restriction. FOXP2-driven network enclosures DCDC2, CDH4, Ezrin-Radixin-Moesin complex, SERPINH1, JAK/STAT signaling,CDH4,DICER1, TARBP2, PIK3K, PIM1, NFE2L2,BACE1, KEAP1, Nrf2 and are important for nervous system development, maintenance, and functioning.

Other signalling pathways affect regulation of receptor-mediated endocytosis AβOs activated p38, mitogen-activated protein kinase, FOX P2 dependent MAPK (Review Wohlgemuth et al, 2014)

and ERK1/2 signalling pathways via the α7nAChR, which in turn results in AβO internalization.

MAPK signalling is implicated downstream of Aβ–PrPC–Fyn Alzheimer’s Amyloid-β oligomers (rescue cellular prion protein induced tau reduction via the Fyn pathway. Mitogen-activated protein kinase signalling pathways are involved in regulating alpha7 nicotinic acetylcholine receptor-mediated amyloid-beta uptake in SH-SY5Y cells, (Yan et al., 2014; Chen et al., 2013)

Also adipokines like adiponectin and leptin are AD-relevant. Adiponectin regulates glucose, lipid and energy metabolism and insulin sensitivity in many tissues via AdipoR1 and -R2 receptors and AMPK, p3-MAPK, PPAR-α and NF-kβ signalling is involved in these processes. (Chandran et al., 2003; Yamauchi et al., 2002; Soodini and Hamdy, 2004)

FoxP2-mi RNA modulation in neurological processes Several studies on songbirds explained how the expression and effects of FOXP2 are influenced by the miRNAs. According to Haesler et al. (2004) and Teramitsu et al. (2004) miRNA expression is indirectly proportional to the FOXP2 level. According to Mohd et al. (2017)intronic miR-3666 modulates different FOXP2 functions such as neuronal growth and development and may contribute to the pathogenesis of schizophrenia and autism. According to Haesler et al. „Incomplete and Inaccurate Vocal Imitation after Knockdown of FoxP2 in Songbird Basal Ganglia Nucleus Area X“ (2007) the reduction of FoxP2 in Area X impaires neuronal dendritic development and learning of singing patterns in young zebra finches. This impairment can be a result of negative miRNAs effect on FOXP2 (Shi et al., 2013). Hessler's group detected with the help of dual luciferase assays, western blotting, Area X tissue dissection, RNA isolation and in situ hybridization that miR-9 and miR-140-5p as well as FoxP2 Expression in Area X was non-linear during vocal learning, so the decline of FoxP2 expression was slow during the growth of the zebra finches, whereas its decline in adult males during undirected vocalisation took place within a few hours. This suggests that mRNA decay does not happen during

transcriptional repression. The researchers proposed a thesis that mRNA decay, induced by the vocalization and mediated by the miRNAs, provides a rapid response to environmental changes, which are necessary for social behaviour. Using lentivirus-mediated RNAi it was possible to prevent accurate song imitation by juveniles. (Haesler et al., 2007; Haeston and White, 2015)

In „Multiple microRNAs regulate human FOXP2 gene expression by targeting sequences in its 3 'untranslated region“ (2014) Fu et al. identified the untranslated UTR3 region of the FOXP2 gene as a regulatory element . Using the microRNAs that interact with this region, they were able to control FOXP2 expression. The FOXP2 mRNA has an approximately 4 kb 3 'untranslated region (3' UTR). It is twice as long as its protein-coding region. This indicates that FOXP2 can be regulated by miRNAs. The expression patterns of let-7a, miR-9 and miR-129-5p in human foetal cerebellum reflect their role in the regulation of FOXP2 expression during early development. These results suggest that various genetic and environmental factors may contribute to speech development. The associated neuronal developmental disorders are influenced, among others, by the miRNA-FOXP2 regulatory network.

Clovis et al. (2012) found that miR-9 and miR-132 could prevent ectopic Foxp2 expression on 3'UTR, which leads to disruption of radial migration in the neocortex of mouse embryos.

HFS diet showed neuroprotective affects, via miR-21 miR-22, miR-34 and miR-101, wich decreases expression of E2F3 and SIRT1 (Kumazaki et al., 2013) , but also via miR-146a, miR-200 and let-7.

Interestingly anti-aging natural products isoflavone, (-)-epigallocatechin-3-gallate, 3,3′-diindolylmethane, indole-3-carbinol, Curcumin positively affects Alzheimer, cardiovascular diseases, atopic asthma, Crohn’s disease, acute and chronic kidney injury, myeloma, glioblastoma, chronic lymphocytic leukaemia, cell lymphoma, osteosarcoma, colo-rectal-, breast-, non-small cell lung cancer and Helicobacter Pylori caused Ulcus. Berberine positively affects hyper-lipemia, cardiovascular diseases, diabetes, colorectal adenoma and Helicobacter pylori caused Ulcus too. It would be interesting to investigate if these effects are congruent with miR expression.

Structure and function of FoxP genes is responsible for his function

Structure of the FoxP2 gene and its isoforms

The FOXP2 gene is located on chromosome 7 and contains at least 280 (according to some data 603) kb, many introns (about 280,000 non-coding base pairs, according to a publication in 2007 - 603,000 base pairs), 7 exons ( 2145 coding base pairs), but their number is variable. (Zhang et al., 2002;Wright and Hastie, 2007)

The protein product of the FOXP2 gene consists of 715 amino acids and has the following domains: a highly conserved DNA-binding domain, a 508 to 584 amino acid "winged" helix domain (BHT) and the forkhead box with highly conserved two beta-sheets, three alpha helixes and a helix-turn-helix-motif-wing.

Structural variations occur between the second and third helix. The polyglutamine-rich regions with the repetitive CAG and CAA sequences show a high mutation rate as well as different length in different taxa. The FOXP2 gene has a zip finger involved in protein-protein interactions and a leucine zipper. The DNA binding in the minor and the major groove to various targets occurs between the third alpha helix (recognition helix) and the second wing of the FOX transcript. (Enard et al., 2002;Kaestner et al., 2000; MacDermot et al., 2005) The hinge loop plays the most important role in the FOXP2 protein binding to the target genes and the mutation P539A changes its form.(Morris et al., 2018)

Alternative splicing creates different FOXP2 isoforms and causes changes in FOXP2 activity. (Castellano and Downward, 2011). Depending on the tissue and cell type, FOXP2 expression can be started on at least 4 starting points (TSSs). (Bruce and Margolis, 2002; Schroeder and Myers, 2008).

Regulation of various genes by dimer formation with FOXP genes

FOXP2 cooperation and FOXP homo- and heterodimers A strong cooperation between FOXP members could be due to the fact that the FOXP2, FOXP1, FOXP3, and FOXP4 are 55-65% identical. A possible explanation for this similarity is proposed by Song et al. in "Genesis of the vertebrate FoxP subfamily member genesis during two ancestral whole genome duplication events" (2016). According to Murugan et al. (2013) decreased FoxP2 expression in the striatal region of adult zebra finches also interferes with their sensitivity to dopaminergic regulation in signalling via D1 receptors in Area-X, they also have co-localized dopamine D1A, D1B and D2 receptors in striatal Foxp2-expressing neurons. Dopamine is considered to be an important neurotransmitter whose deficiency causes some neurodegenerative diseases, e.g. aging relevant Parkinson's disease. This disease is characterized among other things by a washed-out language. It would be interesting to investigate if FOXP2 plays a role in these processes. The FoxP proteins can regulate their target genes in various cellular contexts depending on binding cofactor. Different protein combinations can lead to opposite effects. This could explain why certain tissues might be much more susceptible to the effects of mutations than other tissues. This study opened new perspectives in the regulation of FOXP2 target genes via protein-protein interactions between the FoxP family members, enabling a deeper understanding of the combinatorial control between the FoxP2 and its interaction partners. This study provided new basic knowledge not only about birdsong, but also about the neural function of human speech.

FOXP2 regulates the expression of many genes during embryonic development as well as the WNT and Notch signalling pathways. Further interactions have been observed with some histone family members (H2AFX; H3f3B) and two heat shock proteins (Hsp25; Hsp90a).

Importance of FOXP1/2/4 interaction for oncological processes Several FOXP genes have been observed in many aging relevant oncological processes. Foxp1 / 2/4-NuRD interaction is processed by the p66beta, a transcriptional repressor and a component of NuRD. During this process, the chromatin-remodelling complex regulates gene expression. He also influences the Interleukin-6. Interleukin-6 in turn contributes to the epithelial injury response and activates „myeloid cells“. The „myeloid cells“ are generally associated with cancer and stimulate eg. intestinal cells to divide, which leads to colorectal carcinoma. Artavanis-Tsakonas et al. (1999) studied NCOR2 and SNW1 as part of notch-mediated signalling and its role in proliferation and differentiation processes as well as in apoptosis. The NCOR2 is not only a FOXP2 downstream target but also shows interaction with FOXP1 during myocardial development. (Jepsen et al., 2008), (Wilke et al., 2012) NCOR2-mediated regulation can be considered as a common mechanism by which FOXP1 and other members of the FOXP family regulate gene expression during organogenesis. This study showed different effects of six FOXP1 / 2/4 protein combinations on the NCOR2. The FOXP1 / 2 combination showed the strongest effect. All combinations except the FOXP1 / 4 dimer led to increased NCOR2 expression. The FOXP2 homodimers induced decreased SNW1 expression while the FOXP1 and FOXP4 homodimers led to increased expression. The influence of FOXP1 / 2 and FOXP4 / 2 on SNW1 expression seems to be unlikely. These results gave an interesting insight into the dual FOXP2 function both as a repressor and as an activator. This ability for build different dimer-combinations may be a hint to fine-tune cell-specific transcriptional regulation. The FOXP1 / 2/4 dimer combinations are preferred. These results suggest that relative levels of FOXP1 / 2/4 proteins determine FOXP2's ability to act as an activator or repressor. The researchers found also that FOXP1 / 2, FOXP1 / 4 and FOXP2 / 4 are co-localized.

FOXP2 modulation by alternative splicing The FOXP is also regulated by alternative splicing. This way the FoxP2 gets different isoforms and is homodimerized and this leads to a change of its activity. (Santos et al., 2011) Similar results were reported by Chen et al. (2014) and Olias et al. (2013). This FOXP2 modification was observed in the lower and dorsal thalamus, in the striatum (except Area X) and in the cerebellar Purkinje cells. These are brain areas where the FoxP2 is permanent strongly expressed. (Takahashi et al., 2003), (Ferland et al., 2003), (Haesler et al., 2004) Epigenetic FOXP2 Regulation by Methylation FOXP2 methylation plays an important role by adipositas. Spaeth et al. showed in „The FOXP1, FOXP2 and FOXP4 transcription factors are required for islet alpha cell proliferation and function in mice“ (2015) that Foxp2 is important for the growth and function of pancreatic alfa islands. The islets of Langerhans of the exocrine pancreas contain five different cell populations: the beta, the alpha, the delta, the epsilon and the pancreatic polypeptide cells. These secrete the ghrelin and pancreatic polypeptide hormones (insulin, glucagon, somatostatin). The authors noted that interaction between FOXP2 and FOXP1 / FOXP4 is required for alpha-islet cell proliferation of the mice. Adult beta cells normally secrete insulin, the alpha cells - the glucagon. Autoimmune beta-cell destruction causes type 1 diabetes while type 2 diabetes is characterized by insulin resistance in the peripheral tissues. Type 2 diabetes is accompanied by insulin deficiency and the loss of beta cell identity. The transcription factors accomplish the reprogramming of terminally differentiated cells and embryonic stem cells into the beta-like cells. The members of the FOX superfamily play crucial roles in these processes, which is also aging relevant. E.g., FOXA2 is known as a pancreatic cell fate regulator. FOXM1 controls expression of cycle factors and increases beta cell mass during metabolic stress, including pregnancy stress and partial pancreatectomy. Metformin, Berberine, EGCG, quercetin and other natural products activate cancer relevant AMPK and decrease mTORC1 activity. MTORC1 activity is frequently elevated in CSC including pancreatic CSCs (Ming et al., 2014; Rozengurt et al., 2014; Matsubara et al., 2013) EGCG also positively effects cancer and aging relevant p21Cip1 and negatively the PI3K/PTEN/Akt/mTORC1 pathway (Chen et al., 2012)

According to Alessandro et al., (2015) MET positively influences decreased mitochondrial transmembrane potential, sensitivity to TRAIL via DR5-upregulation, ROS-level and cell cycle in cancer via miR-221 (Tanaka et al., 2015 Matsubara et al., 2013) but also via TRAIL interaction. In this case G-Phase arrest happens via p27Kip-1 and via interaction between caspases (Coleman et al., 2013) Other plant-derived cancer relevant chemicals are e.g. SHH pathway regulating catechins cyclopamine, norcantharidin, sulforaphane and zerumbone (Huang et al., 2013) They act best in combination and suppress ALDH1, MMP-2 and MMP-9 via KRAS under- and let7 miR- upregulation. (Appari et al., 2014) Momordica charantia has positive effect on inflammation and on cancer (Dandawate et al.,2016), e.g. on ovarian cancer via of AMPK up-, mTOR/p70S6K and AKT/ERK/FOXM1 signalling cascade underregulation (Yung et al., 2016) possibly via Alpha-Momorcharin (Deng et al., 2014).

Also Rooperol influences apoptosis with the help of mitochondrial membrane potential. It upregulates ROS via TP53 activation, but also OCT4 and stemness relevant SOX2 and NANOG (Ali et al., 2015) It also increases Pomiferin level, which influences BIM1 and NESTIN (Zhao et al., 2013)

FOXP1, FOXP2 and FOXP4 are of great importance for alpha cell proliferation and function and are expressed in the pancreas and eyes of Xenopus laevis during its development. FOXA1 and FOXA2 also regulate glucagon production and secretion-controlling genes: the MAFB, the Brn4 (also known as the Pou3f4), the PCK2, the Nkx2-2, the Kir6.1 (also known as the Kcnj8), the Sur1 (also known as the ABCC8) and the GIPR.

FOXP2 regulation by external factors Regulation by PH level Blane and Fanucchi (2015) studied in "Effect of pH on the Structure and DNA Binding of the FOXP2 Forkhead Domain." effects of pH from changes 5 to 9 on FOXP2 function and reported that a change in pH (pH 7.5) directly affects the FOXP2 binding affinity via the altered hydrogen bonding pattern. This is due to the protonation or deprotonation of His554 (the amino group of its imidazole side chain, pKa ~ 6.5). The researchers used as methods gel permeation chromatography, ultraviolet circular dichroism, intrinsic and extrinsic fluorescence etc. Their results showed that the pH does not affect the protein secondary structure in the presence or absence of DNA but alters its tertiary structure. The protein showed a less compact structure at low pH in the absence of DNA. When the DNA was added, the protein became more compact, even at low pH and its dimerization potential increased. They regarded the pH as a regulatory mechanism of FOXP2 forkhead domain (FHD) transcription that interacts with the DNA by helix placement in the major groove. These results could also be important in cancer tissue, where FOXP2 expression plays an important role. E.g., not only the genetic component but also a previous chronic gastric ulcer with changed pH plays a significant role in gastric cancer. The gastric ulcer-causing Helicobacter pylori gains an almost pH-neutral environment with the help of the enzyme urease, which splits the urea into carbon dioxide and ammonia. It would be of great scientific interest to investigate whether the change in the gastric pH together with FOXP2 have an effect on the development of gastric cancer.

Regulation through Vitamin-D

Hawes et al. (2015) showed in „Maternal vitamin D deficiency age foetal brain development in the BALB / c mouse“ how the maternal vitamin D deficiency alters foetal brain development in transgenic Balb / c mice. Before and during pregnancy a vitamin D-rich (2.195 IU / kg) or a low (0 IU / kg) diet was given for 5 weeks and the foetal brains were analysed morphologically and for gene expression at 14.5 or 17.5. embryonic day. It was found that the vitamin D deficiency during pregnancy leads to reduction of rump length, lateral ventricle volume and head size. The FoxP2 expression and at the same time the expression of Brain-Derived Neurotrophic Factor (BDNF), the Transforming growth factor-β1 (TGF-β1) and brain tyrosine hydroxylase (TH) in dopaminergic neurons was altered. The vitamin D-poor diet reduced FOXP2 expression in immunoreactive cells and in the developing cortex in female foetuses. These results allowed deeper insight into the medically relevant reasons for foetal degeneration accompanied by prenatal vitamin D deficiency. It is known that vitamin D interacts with neurotrophic factors (NGF, NT3, NT4, GDNF) and its deficiency triggers neurodegenerative diseases in old age. (Jia et al, 2015) It would be of great interest to investigate if this effect is based on the interaction with FOXP2. Since FOXP2 seems to play an important role in many oncological diseases it would be also interesting to investigate if and to what extent this influence is controlled by vitamin D.

FOXP2 regulation by internal factors Regulation by HuR According to Popovitchenko et al. (2016) „Depending on its degree of phosphorylation, the antigen R (HuR) dictates the amount of FOXP2 mRNA and the development of the neocorticals controlled by it, depending on its degree of phosphorylation“ HuR is the main factor in differential translation of autism-associated FoxP subfamily members in the developing neocortex subpopulation of the projection neurons. Regulation by Risperidone and NAP The study of a big Scottish family with severe mental disorders and schizophrenia (Liu et al. 2009) showed a balanced chromosomal translocation [(1:11) (q42.1; q14.3)] and an abnormally-truncated DisC1 protein (a microtubule-regulating and the FOXP2-influenced protein encoded on chromosome 1). The DISC1 is expressed in the cerebral cortex, in the hypothalamus, in the amygdala, in the hippocampal dentate gyrus, in the olfactory bulb and in the cerebellum. The truncated human DISC1 (hDISC1) alters its localization and can no longer interact with microtubules and microtubule-associated proteins. It leads to decreased dendritic growth and branching. Such anomalies have also been found in brain samples from patients with schizophrenia. (Harrison, 2004) This mutation has also been linked to depression and bipolar disorder. (Burdick et al., 2005) A connection between DISC1 and FOXP2 effects human verbal fluency as well as the ability to acquire spoken language. Several studies indicate that FOXP2 polymorphisms are in some cases associated with schizophrenia. (Nicodemus et al., 2014), (Walker et al., 2012), (Lai et al., 2001), (MacDermo et al., 2005), (Sanjuan et al., 2005), (Sanjuan et al. , 2006), (Tolosa et al., 2010) Vaisburd et al. (2015) showed in „Risperidone and NAP protect cognition and normalize gene expression in a schizophrenia mouse model“ that risperidone reduces effects of the DISC1 mutation. Risperidone is an analogue of the microtubule-stabilizing activity-dependent neuroprotective protein (ADNP) with a NAP (NAPVSIPQ) sequence and a SxIP motif (a microtubule junction for microtubule-end binding, responsible for microtubule dynamics and this is also important for synaptic plasticity and neuroprotection). (Oz et al., 2014), (Gozes et al., 2011), (Holtser-Cochav et al., 2006), (Jouroukhin et al., 2013), (Kumar & Wittmann, 2012).

Both risperidone and NAP improved object recognition in the Morris water labyrinth. In contrast to Risperidone, NAP additionally reduced the anxiety in transgenic mice. Doxycycline blocked the expression of the mutated DISC1 gene. The candidate drugs were selected using bioinformatics programs and then affinity chromatographed. Mutations of the DISC1 gene were associated with increased FOXP2 level in hippocampus. FOXP2 levels could be significantly reduced by treatment with NAP, risperidone or their combination. This effect may be due to the blocking of the dopamine and 5HC2A serotonin receptors in the mesolimbic system, which leads to the reduction of negative schizophrenia symptoms. (Meltzer and McGurk, 1999), (Farde et al., 1995)

An increased dopamine level leads to psychotic symptoms. The study by Mendoza et al. (2015), showed that FoxP-expressing neurons in Area-X also contain dopamine receptors 1A, 1B, and 2. Further studies may clarify whether the schizophrenic positive symptoms are due to the dopamine-FOXP2 interaction and how much FOXP 2 can influence dopamine dependent neurodegeneration in aging.

Regulation of FOXP2 and other FOXP genes by SUMOilization

Effect of sumoilization on the FOXP2 gene In „The Key Regulator for Language and Speech Development, FOXP2, is a Novel Substrate for SUMOylation“ Meredith et al. (2016) reported that the FOXP2 is covalently modified on its amino acid residues by both SUMO1 and SUMO3 at phylogenetically conserved lysine 674. The acid residues downstream of the FOXP2 SUMOylation motif are required for full SUMOylation capacity and modulation of its downstream target genes (DISC1, SRPX2 and MiR200c). Sumoylation is a form of posttranslational modification. The SUMOs (small ubiquitin-like modifiers) is a class of regulatory proteins that control the activity of the target protein and its interaction with other proteins. But the modification does not alter the localization and stability of FOXP2. The modulation effect can be reduced by SENP2 (a specific SUMOylation protease). It has also been observed that the human R553H mutation of FOXP2 reduces its SUMOylating ability compared to wild-type FOXP2. Isui et al. (2017) showed in „Sumoylation of FOXP2 Regulated Motor Function and Vocal Communication Through Purkinje Cell“ with the help of in utero electroporation that the SUMOylation of FOXP2 has, among others, an effect on the development of Purkinje cells and plays an important role in motor skills and vocal communication. They identified FOXP2 SUMOylation in the cerebellum of new-born K673 mice with the help of in vivo and in vitro coimmunoprecipitation and concluded that PIAS3 (an E3 ligase of the small ubiquitin-like modifier) catalyses FOXP2 SUMOylation. This SUMOylation modifies the transcription regulation of FOXP2. Estruch et al. (2016) showed in „The language-related transcription factor FOXP2 is post-translationally modified with small ubiquitin-li modifiers“ that post-translational FOXP2 modification is accomplished by members of the PIAS family. These members of the PIAS family are E3 ligases that promote SUMO transfer from the SUMO-conjugating enzyme UBC9 to an acceptor lysine residue of the target protein. (Rytinki et al., 2009) The SUMO2 and the SUMO3 are 95% identical. However, SUMO1 is only to 50% identical to SUMO2 and SUMO3 (Meulmeester and Melchior, 2008). Estruch research group studied various DNA constructs. They used myc-labelled PIAS1 and the His, mCherry and the YFP-labelled SUMOs and the bioluminescence resonance energy transfer assays. For FOXP2 detection they used Western blotting with the anti-V5 antibody and the anti-beta actin as reference. They took UBC9 enzyme to measure the extent of FOXP2 SUMOylation. During the study of FOXP2-PIAS interaction the working group used the BRET assay and the luciferase assays with the Renilla luciferase (Luc). Further methods were the gel-shift assay, the pull-down assay, the yeast two-hybrid assay and the fluorescence microscopy of HEK293 cells. The study revealed that FOXP-2 could be modified by all three human SUMO proteins and that PIAS1 promotes this process. The docking site of SUMO proteins in the FOXP1 and FOXP4 appears to be the most conserved N-terminal region of FOXP2 away from the polyglutamine tract. This region also interacts with the autism-relevant transcription factor TBR1. (Parikshak et al., 2013)

Biochemical aspects of FOXP2 SUMOylation C-terminal carboxyl groups of SUMOS appear to interact with the amino group of FOXP2 via an isopeptide linkage. This amino group of the FOXP2 protein has a lysine side chain with the ΨKX (where Ψ is a hydrophobic amino acid and X is any other amino acid) consensus sequence . The SP-RING domain of the SUMO helps its FOXP2 recognition. N-terminal FOXP2 region contain some of the most important determinants of PIAS binding. These are K674 with PE motif and other residues, e.g. K285, K417 and K560 of the VE sequence. All of these residues are conserved in FOXP2. In this experiment a for SUMO conjugation required Arginine was removed and the SUMOLlyation did not happened. This showed that the K674 is the most important determinant for the SUMO1, SUMO2 and SUMO3 modifications. The mutants assay showed normal dimerization ability in the BRET, so the researchers concluded that the suppressed SUMOylation does not affect the FOXP2 ability to form homodimers or to interact with the CtBP129 co-repressor. This is because FOXP2 dimerization occurs with the help of the leucine zipper domain. However, the leucine zipper domain is not located near the SUMOylation site. This region includes domains that are highly conserved in FOXP1 and FOXP4. These domains serve as recognition sites for the autism-relevant transcription factor TBR1. This area is probably responsible for several protein-protein interactions. (Deriziotis et al., 2014) SUMOilization in FOXP1 / 4 genes Three SUMO proteomic studies of human cell lines identified FOXP1 and FOXP4 as substrates for SUMOilination. (Golebiowski et al., 2009; Tatham et al., 2011; Wen et al., 2014) FOXP1 and FOXP4 contain the SUMOylation relevant DNA motif that is identical to the corresponding FOXP2 motif of the surrounding PE sequence. In BRET study, FOXP1 also showed interactions with PIAS1, PIAS2, and PIAS4, but in contrast to FOXP2, FOXP1 showed little or no interaction with PIAS3. The FOXP1 also showed interactions with all three SUMOs. Probably FOXP1 is also Sumoylated by SUMO1, SUMO2 and SUMO3 at a location, like the K636 site of FOXP2. So, members of the PIAS family are act different in FOXP1 and FOXP2 SUMOylation.

Influence of FOXP2 SUMOlyatin on various diseases It is known that not only FOXP1 and FOXP4 transcription factor TBR1 and co-repressor CtBP1 interact with FOXP2. (Deriziotis et al., 2014; Li et al., 2004; Meulmeester and Melchior, 2008; Rytinki et al., 2009). So by speech and language disorders afflicted family showed a strongly reduced FOXP2 SUMOylation. SUMOylation appears to occur in thousands of nuclear proteins in all cell types at different stages of development (Meulmeester and Melchior, 2008; Gwizdek et al., 2013). According to Bacon and Rappold (2012) and Zhao et al. (2014) SUMO influences dendritic and synaptic morphogenesis by modifying neuronal proteins. Different PIASs may be involved in SUMOylation of various FOXPs and may affect the modification of neuronal circuits. In addition, they support the neuronal plasticity, but it is unclear whether a change in SUMOylation level during brain development leads to functional impairment. It has been demonstrated that PIAS1 knockout causes perinatal lethality in mice. (Loriol et al., 2012; Hasegawa et al., 2014) The study of Estruch et al. (2016) revealed that the four PIAS were continuously expressed but the PIAS1 and PIAS3 were expressed at the same extent in all tissues tested while the PIAS2 and PIAS4 were expressed primarily in testis. Studies of Turner et al. (2013), Adegbola et al. (2015), Nazaryan et al. (2014), Utine et al. (2014) and the fact that the FOXP2 plays a role in neuronal migration, neurite growth, in synaptic plasticity and in the language-related brain circuits (Garcia-Caleroet al., 2015; Tsui et al., 2013; Verne et al., 2011; Groszer et al., 2008) allowed another hypothesis about the cause of speech disorders in the KE family. All members of the by the speech disorder afflicted KE family had a heterozygous R553H mutation in the DNA-binding domain. In twenty other similar cases a missense mutation (R553H) in the FOX DNA binding domain that makes DNA binding and transcriptional repression by FOXP2 impossible was found. Nonsense and frameshift mutations, as well as chromosomal rearrangements were accompanied by similar disabilities. In contrast to the partially compensated interactions between the SUMOS and the K674R mutant the BRET assay showed an almost complete loss of the interactions between the SUMOS and R553H mutant compared to the wild type. The most important SUMOylation site stayed intact in the R553H mutants so the reduction of SUMOylation may base on reduced or absent interaction between the mutant and the PIASs or UBC9. In contrast to the K674R mutants which interact with PIAS but cannot be SUMOylated, the R553H mutants had reduced interaction with PIAS, which can be compensated by PIAS overexpression or by a fusion with UBC9. The R553H mutation also led to partial mislocalization and increased protein aggregation. The R553H mutation may lead to a conformational change that blocks the PIAS binding site. Alternatively, the loss of DNA binding capacity resulting from the R553H mutation and / or destabilization of the protein domain may trigger FOX interactions with other cellular proteins that interfere with PIAS binding. It seems to be useful to investigate the extent of the FOXP2 SUMOylation and the FOXP2-PIAS interactions in different issues and possibly further regulatory mechanisms of aging related neurodegeneration.

FOXP2 regulates proto-oncogenes p21WAF1 / RAS 1, BCL-2, HES1 and other cancer-relevant genes. Gascoyne et al. (2015) demonstrated in „The forkhead transcription factor FOXP2 is required for regulation of p21WAF1 / CIP1 in 143B osteosarcoma cell growth arrest“ with the help of human osteosarcoma cell cultures and normal human osteoblasts that the forkhead transcription factor FOXP2 regulates proto-oncogenes p21WAF1 and CIP1. These proto-oncogenes are required for osteosarcoma cell growth arrest. lncRNA-p21 influences cell proliferation and carcinogenesis via vβ-catenin and JunB. The lncRNA HULC is highly expressed in age related colorectal carcinomas and UCA1 is highly expressed in bladder transitional cell carcinoma by modulating p27 level and can affect cellular senescence.(Parket al., 2011; Smith-Vikos and Slack, 2012) The decrease of growth factor level as well as MAPK reduction induces FOXP2 expression. In turn FOXP2 expression triggers cyclin-dependent kinase inhibitor p21WAF1 / CIP1 expression. It is known that p21 is one of the p53 target genes. (Deiry et al., 1994)19ARF/p53 and p16INK4a/Rb stop cancer cells from proliferation and play an important role in cellular senescence.

During these experiments, the DMEM-cultured and ATCC-derived osteosarcoma cell lines were treated with the IKK pathway inhibitor VII, the MAPK pathway inhibitor, the LY-294002 PI3K pathway inhibitor, the 117082 NF-kappa B pathway inhibitor, the Notch signalling pathway inhibitor or with the DMSO vesicle. Then the alkaline phosphatase and MTS assays were done and 405nm absorbance was measured. Other methods were RNA isolation, cDNA preparation and real-time PCR, protein isolation, immunoblotting, chromatin immunoprecipitation and flow cytometric analysis of the cell cycle distribution. The authors found a link between the 7-time increased FOXP2 expression in murine bone 48 h after treatment and the decreased growth of osteosarcoma cell lines. Unlike mesenchymal tissue osteoblasts did not require RUN involvement in FOXP2 induction. (Zhao et al., 2014) In neuroblastoma cells growth arrest was not accompanied by FOXP2 induction. Cyclin-dependent kinase inhibitors such as p21WAF1, p27KIP1, CIP1CDKN1A and CDKN1B organize CDK inhibition. These experiments also demonstrated that FOXP2 induces CDKN1A / p21 expression in osteoblasts and serves as a primary trigger for cell growth arrest. FOXP2 reduction resulted in strong CDKN1A / p21 and slight CDK4 decline and showed no effect on p53 level. Increased p53 / p21 level had no effect on FOXP-2 level. Immunoprecipitation did not show direct FOXP2 / p21 binding sites and it was demonstrated that although increased FOXP2 expression was accompanied by concomitant IL-6 expression but FOXP2 did not controls p21 expression via IL-6, p21 induction by the FOXP2 takes place indirectly. This is still to be studied. The researchers found a correlation between Lhx8 deficiency factor and FOXP2 expression. There may be a relationship between the FOXP2 and the Soxp2.

Yan X et al. showed 2015 in „Downregulation of FOXP2 promoter human hepatocellular carcinoma cell invasion“ that FOXP2 level in human hepatocellular carcinoma cells is significantly reduced compared to adjacent non-tumor tissue. They used Western blot, transwell assays and immunohistochemistry to measure FOXP2 expression in HCC and in adjacent normal tissues of 50 patients. Their results showed that FOXP2 expression was down-regulated in HCC tumor tissue and that FOXP2-level decrease correlated with poor survival rates. FOXP2 could decrease cell invasion and affect the expression of vimentin and OXE-cadherin.

Since the study by Schroeder and Myers (2008) has emphasized the importance of cell-specific FoxP expression it is especially important to measure it in different tissues and to take a closer look at its cell-specific interaction mechanisms. Chiu et al. (2014) described how FOXP2 inhibits the ERK / MAPK (HSPB7), NOS1, KCNJ15 and SHH pathways (PTCH1) and Vernes et al. (2011) and Ayub et al. (2013) how it influences the oncogene CDK8. FOXP2 effects Bcl-2 expression which seems to be involved in pro-caspase activation and apoptosis. (Devanna et al., 2014). Spiteri et al. (2007) described the cancer-related actin-binding protein genes TAGLN, NOS1, LBR, KCNJ15 and ANK1 as additional FOXP2 target genes. FP2 affects the proto-oncogene C-MET by binding its regulator the AP1-NFAT complex. It also affects the p53 which in turn binds to C-MMET. (Mukamel et al., 2011) (Ho and Crabtree, 2006). MAPK and TGF-β pathways are modified by lncPINT, which is also affected by the with PRC2 interacting p53.

FOXP1 functions, parallels to FOXP2 Like FOXP2's, FOXP1 plays an important role in brain development, because its haploinsufficiency is associated with speech deficits and autism. (Tamura et al., 2003; Le Fevre et al., 2013; Horn et al., 2010; Hamdan et al., 2010; Bacon and Rappold, 2011; Lien et al., 2001; Pariani et al, 2009; Chien et al., 2013; Palmerbo et al., 2013; Carr et al, 2010; Vargha-Khadem et al, 1998; Watkins et al. Brain, 2002; Reimers-Kipping et al. al., 2011; Scharff et Petri, 2011; Li et al., 2015; Song et al., 2015) However, according to „Equivalent missense variant in the FOXP2 and FOXP1 transcription factors causes distinct neurodevelopmental disorders“ (2017) of Sollis et al. the R514H mutation of the FOXP1 gene has a wider and heavier disease manifestation than corresponding p.R553H mutation of the FOXP2 gene. Other functions of the FOXP1 gene were described by Rousso et al. (2016), Wang et al. (2003, 2004), Xing et al.(2017) and Taskapilioglu et al. (2016). According to the recent study by Fröhlich et al. „Foxp1 expression is essential for sex-specific murine neonatal ultrasonic vocalization“ (2017), Foxp1 expression is essential for sex-specific murine neonatal ultrasound vocalization. This study could be of clinical importance because boys are more affected by utism and other speech and speech deficits than girls. They showed that Foxp1 and the androgen receptor are co-expressed in striatal neurons and that brain-specific androgen receptor KO (ArNesCre) mice have a reduced Foxp1 expression in the striatum and an increased Foxp2 level. Oestrogen also reduced the FOXP1 level in breast cancer tissue. (Shigekawa et al., 2011) FOXP3 significance for Alzheimer's disease and for apoptosis Baruch et al. (2015) highlighted the importance of FOXP3 in Alzheimer's disease, in with chronic inflammation leads to the development of myeloid plaques. This inflammation is caused by a loss of Foxp3 + regulatory T cells (Tregs). It would be interesting to investigate whether FOXP3 modulation plays a role in Alzheimer's disease, as it is the case with FOXO1 and FOXO3a. So FOX-miRNA modulation is involved in the progression of Alzheimer's disease. (Lau et al., 2013). This study „Alteration of the microRNA network during the progression of Alzheimer's disease“ observed the hippocampal miRNAs dysregulation of AD-patients. In them miR-132-3p expression on chromosome 17 was altered in both hemispheres. In AD stages III and IV it was severely decreased (41 persons with late-stage AD were compared to 23 healthy volunteers). This dysregulation was mainly observed in tau hyperphosphorylated neurons. This miRNA may affect both the TAU and the transcription factor FOXO1, whose expression is increased in the LOAD hippocampus. According to Wong et al. (2013), miR-132-3p also regulates FOXO3a, which in turn activates pro-apoptotic genes and is overexpressed in LOAD

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