Thanks. I was under the presumption that adding methyl groups can turns genes on as well as off depending on the gene. Looks like that is not the case…
Does anybody know if fever or illness affects the methylation?
Dont get to carried away with that. I felt like shit for the last week or so from the flu.
I have not read much about this subject yet, but I think we need to test this hypothesis. For me it would be interesting to know if this hypothesis explains some of the short-time recovery effects.
Could someone explain why we suppose that our problem is over-methylation and not under-methylation?
As far as i understand this, is our theory based on this paper(posted by awor):
I can’t read the whole theses, but in the abstract you can find this sentence:
“For example the antihypertensive hydralazine inhibits DNA methylation”
Why shouldn’t fin also inhibts DNA methylation? Which would mean we suffer from under-methylation.
Epigenetic setting for long-term expression of estrogen receptor α and androgen receptor in cells
Epigenetic regulation of the nuclear estrogen and androgen receptors, ER and AR, constitutes the molecular basis for the long-lasting effects of sex steroids on gene expression in cells. The effects prevail at hundreds of gene loci in the proximity of estrogen- and androgen-responsive elements and many more such loci through intra- and even inter-chromosomal level regulation. Such a memory system should be active in a flexible manner during the early development of vertebrates, and later replaced to establish more stable marks on genomic DNA. In mammals, DNA methylation is utilized as a very stable mark for silencing of the ERα and AR isoform expression during cancer cell and normal brain development. The factors affecting the DNA methylation of the ERα and AR genes in cells include estrogen and androgen. Since testosterone induces brain masculinization through its aromatization to estradiol in a narrow time window of the perinatal stage in rodents, the autoregulation of estrogen receptors, especially the predominant form of ERα, at the level of DNA methylation to set up the “cell memory” affecting the sexually differentiated status of brain function has been attracting increasing attention. The alternative usage of the androgen-AR system for brain masculinization and estrogenic regulation of AR expression in some species imply that the DNA methylation pattern of the AR gene can be established by closely related but different systems for sex steroid-induced phenomena, including brain masculinization.
Androgen Receptor Mutations Are Associated with Altered Epigenomic Programming as Evidenced by HOXA5 Methylation
Male external genital differentiation is accompanied by implementation of a long-term, male-specific gene expression pattern indicating androgen programming in cultured genital fibroblasts. We hypothesized the existence of an epigenetic background contributing to this phenomenon. DNA methylation levels in 2 normal scrotal fibroblast strains from 46,XY males compared to 2 labia majora fibroblast strains from 46,XY females with complete androgen insensitivity syndrome (AIS) due to androgen receptor (AR) mutations were analyzed by Illumina GoldenGate methylation arrays®. Results were validated with pyrosequencing in labia majora fibroblast strains from fifteen 46,XY patients and compared to nine normal male scrotal fibroblast strains. HOXA5 showed a significantly higher methylation level in complete AIS. This finding was confirmed by bisulfite pyrosequencing of 14 CpG positions within the HOXA5 promoter in the same strains. Extension of the 2 groups revealed a constant low HOXA5 methylation pattern in the controls in contrast to a highly variable methylation pattern in the AIS patients. HOXA5 represents a candidate gene of androgen-mediated promoter methylation. The constantly low HOXA5 DNA methylation level of normal male scrotal fibroblast strains and the frequently high methylation levels in labia majora fibroblast strains in AIS indicate for the first time that androgen programming in sexual differentiation is not restricted to global gene transcription but also occurs at the epigenetic level.
Metabolomic Profiling Reveals a Role for Androgen in Activating Amino Acid Metabolism and Methylation in Prostate Cancer Cells
Prostate cancer is the second leading cause of cancer related death in American men. Development and progression of clinically localized prostate cancer is highly dependent on androgen signaling. Metastatic tumors are initially responsive to anti-androgen therapy, however become resistant to this regimen upon progression. Genomic and proteomic studies have implicated a role for androgen in regulating metabolic processes in prostate cancer. However, there have been no metabolomic profiling studies conducted thus far that have examined androgen-regulated biochemical processes in prostate cancer. Here, we have used unbiased metabolomic profiling coupled with enrichment-based bioprocess mapping to obtain insights into the biochemical alterations mediated by androgen in prostate cancer cell lines. Our findings indicate that androgen exposure results in elevation of amino acid metabolism and alteration of methylation potential in prostate cancer cells. Further, metabolic phenotyping studies confirm higher flux through pathways associated with amino acid metabolism in prostate cancer cells treated with androgen. These findings provide insight into the potential biochemical processes regulated by androgen signaling in prostate cancer. Clinically, if validated, these pathways could be exploited to develop therapeutic strategies that supplement current androgen ablative treatments while the observed androgen-regulated metabolic signatures could be employed as biomarkers that presage the development of castrate-resistant prostate cancer.
After the inconclusive results of the recent research effort, is hypermethylation still a possible explanation of our problems?
Yes. The next step i imagine is to try and pinpoint where along the sequence of events from androgen binding to receptor is the problem. Is it epigenetic (methylation, histone acetylation), is it after this and more about proteins (post translational modifications) and signalling, is it secondary to another pathology such as autoimmune conditions or infection?
There has been some breakthoughs recently with the study of misfolded proteins in HD and AIDS if that is part of the problem.
We cannot be insensitive to androgen (no matter what the cause may be). If that was the case, then:
- why do most of us have low T? The hypothalamus should sense little T and ask for more.
- why can somebody grow muscles on TRT/clomid (me, moonman1)?
- why are some people growing excessive body hair or having an increase in hair loss? If your answer is that the receptors are hypersensitive, then why don’t we have a hyperlibido?
- why has never awor replied to question 1, which I asked several times to him in different threads?
In general terms we are suffering from various degrees of androgen insensitivity. I personally took over twice the normal amount of Depo T over several months and felt nothing. Jacobs has said he had someone over 4000 total T and he too was still hypogonadal. Personally I cannot grow muscle were it was very easy to do so prior to Fin and some muscle groups seem to atrophy faster than others.
The answers to questions 1-4 is I have no idea. Maybe you and moonman arnt quite as androgen insensitive as the rest of us for whatever reason. For some reason you can compensate in those areas when others of us can’t. I agree that is is certainly likely there are several issues at play or maybe some mechanism we don’t even uderstand yet. Thankfully a biological investigation is underway and I hope we all do everything we can to encourage it and further research.
Awor chose a logical place to begin an investigation. I don’t recall him ever saying this would be a perfect theory or there that there are no other issues at play. I really don’t understand everyone’s need to attack him. Since you are in academia I’m sure you also well understand it’s a very rare occasion when a theory in turns out to be perfectly correct.
We are not. We cannot possibly be androgen insensitive, otherwise we would have high testosterone, per point 1 above. And don’t tell me that the receptors are in fact hypersensitive, otherwise we would have high libido.
This doesn’t prove that there is something wrong with our androgen receptors. As awor himself found out, other hormones (for example, T3) determine how easily androgens can enter receptors.
I’m probably more “sensitive” than most of the people here for the simple reason that I’m supplementing with T4. Before supplementing with T4, I was indeed responding negatively to androgens (see my story).
It’s not logical, as it contradicts point 1 above.
I’m attacking the attitude of getting fixated with the investigation of androgen receptors, until somebody can provide an answer to my questions.
Yes, but when a theory contradicts an observation in the real world, the theory needs to be changed.
My T has almost doubled since crash. I have had total T scores in the 700’s and free T in the 150 range and still had absolutely no changes in symptoms.
I did a T3 cycle per Shippen and had no results personally. I did this about the same time as a clomid cycle. Per my personal experience it hasnt worked.
If you have another theory I think that’s great. I don’t think Awor is being sarcastic at all when he says “go pursue research”. More eyes on this the better IMO.
As Boston332 has said, no one is stopping you if you want to pursue other angles, but i don’t get people who are actively discouraging the study of PFS at the molecular level. Wouldn’t you at least want to eliminate the possibility?
No one should be so arrogant as to claim with absolute certainty that they know exactly what the underlying problem of PFS is. All awor is trying to do is investigate an angle that he, and others believe could explain PFS, through scientific research. If it turns out that this is not the case then we have eliminated another possibility, nothing more nothing less. Either way we are better off for having investigated are we not?
I don’t think the degree of low T we have explains extreme penile shrinkage, muscle loss, weight gain and so on. In most cases we do not know what level of T most of us began with. I know my total T and free T have more than doubled over the more than year and a half I’ve been off Fin. Not to mention the time I was on TRT. Still, I have absolutely no resolution of side effects. Still have penile tissue changes, muscle waste, fat gain, etc.
As awor correctly said, we (as a community) should undertake 2 types of effort. One is a research effort organized together with a lab; the other is an approach based on treatments that our members try out. Any hopeless research would encourage people to uselessly wait for the results of this research, while it would be more productive if they undertook promising hormonal treatments and shared their experience. Similarly, I discourage undertaking hopeless hormonal treatments that will never work, such as going on TRT alone, or taking arimidex alone. The reason why these treatments should be discouraged is that seeing somebody for whom TRT doesn’t work won’t add anything to our knowledge. On the other hand, sharing the results of a treatment that nobody has tried, or a treatment that has lead to a recovery, etc would enrich our knowledge. In other words, I think that every hopeless avenue should be discouraged to increase our “efficiency”.
The possibility is already eliminated. If our receptors responded poorly to androgen, then we wouldn’t have members with low T, low DHT, and low E2. (unless you have a different explanation)
huh… where do I make such claim?
Not to speak for Mario but I think he means “no one” as in no one on this board/those working on PFS not you in particular.
I don’t believe anyone will put off treatment or will tend to put off treatment due to the current research. I’ll be seeing Shippen for my yearly in a few weeks and will be seeing a naturopath/internist for a homeopathic approach to work together with whatever new ideas the PFS docs can come up with. I’m just hoping the docs will take any published research findings into account.
Yes, i didn’t mean you specifically m_81.
Whether or not you think awor’s research is hopeless is a matter of opinion and you are entitled to yours. I am not going to go back and forth arguing the merits of different theories as we have both read enough on this forum to have made up our own minds.
In saying that i think that you would agree that we are all old enough to think for ourselves, and the idea that we should avoid scientific research (once again personal opinions on it’s validity aside) because it might stop people from going to see a doctor is a little ridiculous, especially given that from what i have seen doctors have had extremely limited success in treating PFS anyway (even that is being rather generous).
The bottom line for me is that awor’s work might be the one and only chance we get to investigate this at the molecular level, hormonal therapy is there for people to try now if they want to and will be there in the future but if we let this study pass then we miss an opportunity to get valuable insight.
Nearly all the recoveries from PFS have been natural recoveries or dumb luck. There are still many unanswered questions. If awor’s research can answer even a few of those, or even eliminate a few possibilities then it is worth the effort in my opinion. Ask yourself, are we honestly making that much progress on our own that we can afford to turn away help? I don’t think so.
How would a demethylase agent differ from a Methyltransferase inhibitor when it comes to treating our case?
I just wanted to add that I am a big fan of demethylating agents, and wanted to correct the post that said that Dacogen was Azacytidine (it’s actually Vidaza).