13 replies to this topic

[cylack]

Saw this article in The Economist talking about this gene KL, particularly the KL-VS variant, that had a strong link to intelligence. The protein product of the gene is called klotho protein. Anyone have any idea what foods or supps could upregulate this gene or the protein product?

 

http://www.economist...y-has-just-been

[Blankspace]

Here is the full text:

http://www.cell.com/...1247(14)00287-3

 

The SNP:

Rs9536314

 

Snpedia Klotho:

http://www.snpedia.com/index.php/KL

 

 

[LexLux]

Its also worth mentioning that this is actually also a longevity gene. Interestingly it increases "...brain skills such as thinking, learning and memory regardless of their age, sex, or whether they have a genetic risk factor for Alzheimer's disease".

 

So far I've only found some associations with HDAC inhibitor trichostatin A and DNA demethylating agent 2'-deoxy-5-azacytidine. I am not advising anyone to try these, I'm merely interested in exploring this from an academic perspective. 

 

1) The anti-aging gene KLOTHO is a novel target for epigenetic silencing in human cervical carcinoma - 

 

"To determine whether the loss of KLOTHO expression is associated with epigenetic gene silencing, we used RT-PCR to examine transcriptional reactivation in representative cell lines lacking KLOTHOmRNA following treatment with either the DNA methyltransferase (DNMT) inhibitor 2'-deoxy-5-azacytidine (DAC) or the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) (Figure 1C). DNA demethylation by DAC induced significant restoration of KLOTHO mRNA expression in the SNU-1299 cell line, but incomplete upregulation was seen in the SiHa cell line. When each cell line was treated with TSA alone, KLOTHO restoration was detected only in the SiHa cell line. A synergistic effect of DAC and TSA was observed in the SNU-1299 cell line, in which the KLOTHOmRNA was restored only with a DNA demethylating drug. Taken together, these data imply that the downregulation of KLOTHO in cervical cancer cell lines is correlated with epigenetic inactivation mechanisms involving DNA methylation and histone deacetylation."

 

[...]

"Histone deacetylation is the major epigenetic silencing mechanism for KLOTHO in the SiHa cell line

 

A chromatin immunoprecipitation (ChIP) assay using anti-acetyl histone H3 (AcH3) and H4 (AcH4) antibodies was performed to determine whether SiHa cells utilize local histone modification as a mechanism of KLOTHO silencing. The immunoprecipitated DNA was analyzed by PCR to elucidate the histone H3 and H4 acetylation level of the KLOTHO promoter region. Figure 2A shows the promoter region around the transcription start site (+1) that was analyzed by the ChIP assay. As shown in Figure 4, marked differences were observed between the acetylation level of histones H3 and H4 in untreated SiHa cells. Although very low levels of PCR products were detected in the control SiHa cells, TSA treatment resulted in a dramatic amplification of specific DNA fragments immunoprecipitated with the AcH3 antibody. This result suggested that the KLOTHO promoter is enriched in deacetylated histone H3, leading to repressive histone modification in the SiHa cell line. On the other hand, the acetylation level of histone H4 seemed to not be associated with restoration of KLOTHO mRNA after treatment with TSA in the SiHa cell line. Suzuki et al. reported that a group of genes lacking promoter CpG methylation tended to be reactivated after HDAC inhibition alone[20]. Our study identified unmethylated CpGs present in the KLOTHO promoter in the SiHa cell line (Figure 3). Thus, deacetylated histone H3 in the promoter region is strongly correlated with epigenetic silencing of KLOTHO in the SiHa cell line."

 

2) http://www.biomedcen...-4598-9-109.pdf -

 

Abstract

Background: Klotho was originally characterized as an anti-aging gene that predisposed Klotho-deficient mice to a 

premature aging-like syndrome. Recently, KLOTHO was reported to function as a secreted Wnt antagonist and as a 

tumor suppressor. Epigenetic gene silencing of secreted Wnt antagonists is considered a common event in a wide 

range of human malignancies. Abnormal activation of the canonical Wnt pathway due to epigenetic deregulation of 

Wnt antagonists is thought to play a crucial role in cervical tumorigenesis. In this study, we examined epigenetic 

silencing of KLOTHO in human cervical carcinoma.

 

Results:

Loss of KLOTHO mRNA was observed in several cervical cancer cell lines and in invasive carcinoma samples, 

but not during the early, preinvasive phase of primary cervical tumorigenesis. KLOTHO mRNA was restored after 

treatment with either the DNA demethylating agent 2'-deoxy-5-azacytidine or histone deacetylase inhibitor 

trichostatin A. Methylation-specific PCR and bisulfite genomic sequencing analysis of the promoter region of KLOTHO 

revealed CpG hypermethylation in non-KLOTHO-expressing cervical cancer cell lines and in 41% (9/22) of invasive 

carcinoma cases. Histone deacetylation was also found to be the major epigenetic silencing mechanism for KLOTHO in 

the SiHa cell line. Ectopic expression of the secreted form of KLOTHO restored anti-Wnt signaling and anti-clonogenic 

activity in the CaSki cell line including decreased active β-catenin levels, suppression of T-cell factor/β-catenin target 

genes, such as c-MYC and CCND1, and inhibition of colony growth.

 

Conclusions:

Epigenetic silencing of KLOTHO may occur during the late phase of cervical tumorigenesis, and 

consequent functional loss of KLOTHO as the secreted Wnt antagonist may contribute to aberrant activation of the 

canonical Wnt pathway in cervical carcinoma.

Edited by LexLux, 10 May 2014 - 07:22 PM.

[blood]

ARBs like valsartan have been shown to increase Klotho levels in people (the studies were done with diabetics, from memory).

ARB use has been linked to reduced incidence of Alzheimer's (observational/epidemiological studies).

Some ARBs have been shown to have mild cognitive enhancing properties in healthy subjects.

Apologies for lack of references, posting from bed on my ipad.

Edited by blood, 10 May 2014 - 07:18 PM.

[Mr Matsubayashi]

I'm a (G:T) so on the better end alas not the best (G:G).

 

A further target for future gene therapy

[Blankspace]

I'm a (G:T) so on the better end alas not the best (G:G).

 

A further target for future gene therapy

 

It looks like (G:T) is superior to both of the homozygous alleles, with (G:G) coming out as the worst of the three, at least as far as lifespan is concerned.

 

Life Extension Factor Klotho Enhances Cognition (Results):

"Twenty-six percent of individuals were heterozygous for the KL-VS allele, slightly above typical frequencies of 20%–25% (Arking et al., 2002). Three percent were homozygous for KL-VS, a rare genotype that for unknown reasons is associated with decreased lifespan and detrimental effects Arking et al., 2002, Arking et al., 2005, Deary et al., 2005); they were excluded from the study."

Edited by Blankspace, 11 May 2014 - 02:07 PM.

[Phoenicis]

Shame there aren't more drugs that induce klotho?

[Phoenicis]

 

 

1) The anti-aging gene KLOTHO is a novel target for epigenetic silencing in human cervical carcinoma - 

 

"To determine whether the loss of KLOTHO expression is associated with epigenetic gene silencing, we used RT-PCR to examine transcriptional reactivation in representative cell lines lacking KLOTHOmRNA following treatment with either the DNA methyltransferase (DNMT) inhibitor 2'-deoxy-5-azacytidine (DAC) or the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) (Figure 1C). DNA demethylation by DAC induced significant restoration of KLOTHO mRNA expression in the SNU-1299 cell line, but incomplete upregulation was seen in the SiHa cell line. When each cell line was treated with TSA alone, KLOTHO restoration was detected only in the SiHa cell line. A synergistic effect of DAC and TSA was observed in the SNU-1299 cell line, in which the KLOTHOmRNA was restored only with a DNA demethylating drug. Taken together, these data imply that the downregulation of KLOTHO in cervical cancer cell lines is correlated with epigenetic inactivation mechanisms involving DNA methylation and histone deacetylation."

 

[...]

"Histone deacetylation is the major epigenetic silencing mechanism for KLOTHO in the SiHa cell line

 

A chromatin immunoprecipitation (ChIP) assay using anti-acetyl histone H3 (AcH3) and H4 (AcH4) antibodies was performed to determine whether SiHa cells utilize local histone modification as a mechanism of KLOTHO silencing. The immunoprecipitated DNA was analyzed by PCR to elucidate the histone H3 and H4 acetylation level of the KLOTHO promoter region. Figure 2A shows the promoter region around the transcription start site (+1) that was analyzed by the ChIP assay. As shown in Figure 4, marked differences were observed between the acetylation level of histones H3 and H4 in untreated SiHa cells. Although very low levels of PCR products were detected in the control SiHa cells, TSA treatment resulted in a dramatic amplification of specific DNA fragments immunoprecipitated with the AcH3 antibody. This result suggested that the KLOTHO promoter is enriched in deacetylated histone H3, leading to repressive histone modification in the SiHa cell line. On the other hand, the acetylation level of histone H4 seemed to not be associated with restoration of KLOTHO mRNA after treatment with TSA in the SiHa cell line. Suzuki et al. reported that a group of genes lacking promoter CpG methylation tended to be reactivated after HDAC inhibition alone[20]. Our study identified unmethylated CpGs present in the KLOTHO promoter in the SiHa cell line (Figure 3). Thus, deacetylated histone H3 in the promoter region is strongly correlated with epigenetic silencing of KLOTHO in the SiHa cell line."

 

 

 

 

Don't go taking any of these before seeking independent medical advice since I'm not qualified to give any. Tributyrin should be studied further since it works very similar to trichostatin A as mentioned earlier. Can anyone point out the downsides of tributyrin specifically?

 

People have noted how quickly butyrate is metabolised, but it seems tributyrin has much better pharmokinetics and the other interesting thing is that tributyrate is sold as a food grade flavor?!  10kg for 200 British pounds? Am I mistaken? Apparently it's naturally present in butter as well.

 

Sodium butyrate stimulates expression of fibroblast growth factor 21 in liver by inhibition of histone deacetylase 3.

Li H1, Gao Z, Zhang J, Ye X, Xu A, Ye J, Jia W.

Abstract

 

Fibroblast growth factor 21 (FGF21) stimulates fatty acid oxidation and ketone body production in animals. In this study, we investigated the role of FGF21 in the metabolic activity of sodium butyrate, a dietary histone deacetylase (HDAC) inhibitor. FGF21 expression was examined in serum and liver after injection of sodium butyrate into dietary obese C57BL/6J mice. The role of FGF21 was determined using antibody neutralization or knockout mice. FGF21 transcription was investigated in liver and HepG2 hepatocytes. Trichostatin A (TSA) was used in the control as an HDAC inhibitor. Butyrate was compared with bezafibrate and fenofibrate in the induction of FGF21 expression. Butyrate induced FGF21 in the serum, enhanced fatty acid oxidation in mice, and stimulated ketone body production in liver. The butyrate activity was significantly reduced by the FGF21 antibody or gene knockout. Butyrate induced FGF21 gene expression in liver and hepatocytes by inhibiting HDAC3, which suppresses peroxisome proliferator-activated receptor-? function. Butyrate enhanced bezafibrate activity in the induction of FGF21. TSA exhibited a similar set of activities to butyrate. FGF21 mediates the butyrate activity to increase fatty acid use and ketogenesis. Butyrate induces FGF21 transcription by inhibition of HDAC3.

 

 

Tributyrin, a Stable and Rapidly Absorbed Prodrug of Butyric Acid, Enhances Antiproliferative Effects of Dihydroxycholecalciferol in Human Colon Cancer Cells

 

 Tanja Gaschott, Dieter Steinhilber*, Vladan Milovic, and Jürgen Stein2

 

"Butyrate, a normal constituent of the colonic luminal contents, is formed by bacterial fermentation of unabsorbed complex carbohydrates in the mammalian digestive tract. In normal colonic mucosa, butyrate serves as a primary energy source, promotes growth of normal colonic epithelial cells in vivo and in vitro and plays a role in preventing certain types of colitis (1). In contrast, in a wide variety of neoplastic cells, butyrate acts as a potent antineoplastic agent, i.e., it inhibits growth and induces differentiation, restoring normal phenotype and function (2). The studies done during the last decade provide multiple lines of evidence that butyrate indeed interferes with the pathogenesis of colorectal cancer. Butyrate inhibits DNA synthesis and arrests growth of neoplastic colonocytes in G1 (3), modifies expression of genes involved in chemotherapy resistance (4) and in cell proliferation/differentiation (5, ,6), and induces apoptosis by a p53-independent pathway (7). At least some of butyrate?s antineoplastic effects in colon cancer cells may be due to its synergistic action with another antiproliferative agent, 1,25-dihydroxyvitamin D3 [dihydroxycholecalciferol; (OH)2D3]. In various cancer cell lines it has been shown that butyrate and (OH)2D3 act synergistically in reducing proliferation and enhancing differentiation of neoplastic cells (8, 9, 10).

 

In spite of its early promise, butyrate is not among the drugs used for cancer treatment. The major problem has been to achieve and maintain its millimolar concentrations in blood. Butyrate is metabolized rapidly as soon as it enters the colonocyte via its active transport system (11, 12, 13), and its plasma concentrations are far below those required to exert its antiproliferative/differentiating actions.

 

A prodrug of natural butyrate, tributyrin, is a neutral short-chain fatty acid triglyceride that is likely to overcome the pharmacokinetic drawbacks of natural butyrate as a drug (14). Because it is rapidly absorbed and chemically stable in plasma, tributyrin diffuses through biological membranes and is metabolized by intracellular lipases, releasing therapeutically effective butyrate over time directly into the cell. Compared with butyrate, tributyrin has more favorable pharmacokinetics (14, 15, 16) and is well tolerated (17). Liquid tributyrin filled into gelatin capsules and administered orally resulted in millimolar concentrations of butyrate both in plasma and inside the cell (17). In vitro, tributyrin has potent antiproliferative, proapoptotic and differentiation-inducing effects in neoplastic cells (18, 19, 20). In this study, human colon cancer cells (Caco-2) were used to investigate the effects of tributyrin on growth and differentiation."

 

 

CLINICAL TRIALS -

 

1) Phase I study of the orally administered butyrate prodrug, tributyrin, in patients with solid tumors.

 

Butyrates have been studied as cancer differentiation agents in vitro and as a treatment for hemoglobinopathies. Tributyrin, a triglyceride with butyrate molecules esterified at the 1, 2, and 3 positions, induces differentiation and/or growth inhibition of a number of cell lines in vitro. When given p.o. to rodents, tributyrin produces substantial plasma butyrate concentrations. We treated 13 patients with escalating doses of tributyrin from 50 to 400 mg/kg/day. Doses were administered p.o. after an overnight fast, once daily for 3 weeks, followed by a 1-week rest. Intrapatient dose escalation occurred after two courses without toxicity greater than grade 2. The time course of butyrate in plasma was assessed on days 1 and 15 and after any dose escalation. Grade 3 toxicities consisted of nausea, vomiting, and myalgia. Grades 1 and 2 toxicities included diarrhea, headache, abdominal cramping, nausea, anemia, constipation, azotemia, lightheadedness, fatigue, rash, alopecia, odor, dysphoria, and clumsiness. There was no consistent increase in hemoglobin F with tributyrin treatment. Peak plasma butyrate concentrations occurred between 0.25 and 3 h after dose, increased with dose, and ranged from 0 to 0.45 mM. Peak concentrations did not increase in three patients who had dose escalation. Butyrate pharmacokinetics were not different on days 1 and 15. Because peak plasma concentrations near those effective in vitro (0.5-1 mM) were achieved, but butyrate disappeared from plasma by 5 h after dose, we are now pursuing dose escalation with dosing three times daily, beginning at a dose of 450 mg/kg/day.

 

2) Clinical and pharmacologic study of tributyrin: an oral butyrate prodrug

 

Martin J. Edelman, Kenneth Bauer, Shyam Khanwani, Nancy Tait, Jane Trepel, Judith Karp, Noble Nemieboka, Eun-Joo Chung, David Van Echo

 

Purpose

Butyrate is a small polar compound able to produce terminal differentiation and apoptosis in a variety of in vitro models at levels above 50?100 ?M. Previously our group demonstrated that daily oral administration of the prodrug, tributyrin, is able to briefly achieve levels >100 ?M. Given in vitro data that differentiating activity requires continuous butyrate exposure, the short t1/2 of the drug and a desire to mimic the effects of an intravenous infusion, we evaluated a three times daily schedule.

 

Patients and methods

Enrolled in this study were 20 patients with advanced solid tumors for whom no other therapy was available, had life expectancy greater than 12 weeks, and normal organ function. They were treated with tributyrin at doses from 150 to 200 mg/kg three times daily. Blood was sampled for pharmacokinetic analysis prior to dosing and at 15 and 30 min and 1, 1.5, 2, 2.5, 3, 3.5 and 4 h thereafter.

 

Results

The patients entered comprised 15 men and 5 women with a median age of 61 years (range 30?74 years). Prior therapy regimens included: chemotherapy (median two prior regimens, range none to five), radiation therapy (one), no prior therapy (one). There was no dose-limiting toxicity. Escalation was halted at the 200 mg/kg three times daily level due to the number of capsules required. A median butyrate concentration of 52 ?M was obtained but there was considerable interpatient variability. No objective responses were seen. There were four patients with prolonged disease stabilization ranging from 3 to 23 months; median progression-free survival was 55 days. Two patients with chemotherapy-refractory non-small-cell lung cancer had survived for >1 year at the time of this report without evidence of progression.

 

Conclusion

Tributyrin is well tolerated and levels associated with in vitro activity are achieved with three times daily dosing.

 

 

3) No results posted here? http://www.clinicalt...ibutyrin&rank=1

 

 

  Anyone know enough about sirtuins to interpret this?:

 

Differential regulation of the Sir2 histone deacetylase gene family by inhibitors of class I and II histone deacetylases.

Kyrylenko S1, Kyrylenko O, Suuronen T, Salminen A.

Abstract  

The Sir2 histone deacetylase gene family consists of seven mammalian sirtuins (SIRTs) which are NAD-dependent histone/protein deacetylases. Sir2 proteins regulate, for instance, genome stability by chromatin silencing in yeast. In mammals, their function is still largely unknown. Due to the NAD+ dependency, Sir2 might be the link between metabolic activity and histone/protein acetylation. Regulation of gene expression also seems to play an important role in Sir2 functions, since increasing the dosage of Sir2 genes increases genome stability in yeast and Caenorhabditis elegans. We observed that the modification of histone/protein acetylation status by several class I and II histone deacetylase (HDAC) inhibitors induces differential changes in gene expression profiles of seven SIRT mRNAs in cultured neuronal cells. SIRT2, SIRT4 and SIRT7 were upregulated, whereas SIRT1, SIRT5 and SIRT6 were downregulated by trichostatin A (TSA) and n-butyrate. The upregulation of SIRT mRNAs was inhibited by actinomycin D. Interestingly, the regulation of SIRT mRNAs was highly similar both in mouse Neuro-2a neuroblastoma cells and post-mitotic rat primary hippocampal and cerebellar granule neurons. Using a chromatin immunoprecipitation technique, we showed that the upregulation of SIRT2 expression with TSA is related to the hyperacetylation of DNA-bound histone H4 within the first 500 bp upstream of the transcription start site of the SIRT2 gene. Chemically different types of HDAC inhibitors, such as TSA, apicidin, SAHA, M344 and n-butyrate induced remarkably similar responses in SIRT1-7 mRNA expression patterns. Differential responses in SIRT mRNA expression profiles indicate that the expression of the Sir2 family of genes is selectively regulated and dependent on histone/protein acetylation status.

 

 

 

 

Edited by Phoenicis, 15 May 2014 - 02:10 PM.

[Phoenicis]

Hypothesized MoA:

 

Tributyrin -> Histone H3 Acetylation -> increase of KL expression (dependant on genotype) -> increased KLOTHO protein

Edited by Phoenicis, 15 May 2014 - 06:47 PM.

[therein]

Looks like I am T:T on Rs9536314. SNPedia says that's the "worst" but other sources say it's the "best" lifespan and intelligence-wise. While I do realize that these don't mean much, it is just one of the millions of genes that regulate intelligence, is T:T the best one or the worst one?

[tazzz96]

Looks like I am T:T on Rs9536314. SNPedia says that's the "worst" but other sources say it's the "best" lifespan and intelligence-wise. While I do realize that these don't mean much, it is just one of the millions of genes that regulate intelligence, is T:T the best one or the worst one?

I'm a (G:T) so on the better end alas not the best (G:G).
 
A further target for future gene therapy

How do you guys know that you're G:T or T:T or whatever?

Sent from my Nexus 4 using Tapatalk

[therein]

 

Looks like I am T:T on Rs9536314. SNPedia says that's the "worst" but other sources say it's the "best" lifespan and intelligence-wise. While I do realize that these don't mean much, it is just one of the millions of genes that regulate intelligence, is T:T the best one or the worst one?

 

I'm a (G:T) so on the better end alas not the best (G:G).
 
A further target for future gene therapy

How do you guys know that you're G:T or T:T or whatever?

Sent from my Nexus 4 using Tapatalk

 

 

The 23andMe test's raw dump

[LexLux]

This is just my humble non-medical research, get independent medical advice if anything - 

 

Another HDAC inhibitor  - beta-hydroxybutyrate, this recent publication has noted that the detrimental effects of HDAC1 inhibition are offset by SIRT1 activation (see pp.49 of the free full text). Similarly to trichostatin A, this HDACi actually also inhibits HDAC3, but it is less potent than butyrate. Tributyrin could be the among the most potent natural HDACis, but I'm not a doctor and cannot comment on it's safety. 

 

Could these then raise KLOTHO? This may require further study.

 

Beta-hydroxybutyrate is actually a ketone and can be raised by calorie restriction, fasting, and by medium chain triglycerides. This ketone was traditionally thought of as an alternative source of energy for the brain when glucose levels are low, but has recently been found to act as a signalling molecule and as an HDAC inhibitor.

 

A mere 30g/d of MCTs produces a mild ketosis, similar to calorie restriction, one could also reasonably expect similar effects. Aside from more energy and less appetite, these would be -

 

(Ref: John C. Newman, Ketone bodies as signaling metabolites, Trends Endocrinol Metab. 2014 Jan;25(1):42-52. [Be sure to see the full text])

• lower mTOR activity
• less IGF-1 signalling
• more AMPK activity
• Upregulation of FOXO3
• more protein acetylation
• more stress resistance

In Addition - 

• possibly better cognitive function, see also this
• substantial protection against oxidative stress
• It also inhibits HDAC1 which is over-expressed in psoriasis and apparently the detrimental effects of HDAC1 inhibition are offset by SIRT1 activation (see pp 49 of John Newman Article) .
•  

Edited by LexLux, 02 June 2014 - 09:08 PM.

[Phoenicis]

Resveratrol Increases Klotho Gene Expression see this thread - 

 

http://www.longecity...ene-expression/