Restoring androgen sensitivity with epigenetic modulators

Hi guys. It’s been a while since I’ve posted. I’ve been through a couple job transitions and haven’t had as much time to devote to this as I wanted. Anyway, I’ve been thinking a lot about the issue and wanted to bring up some points for discussion (sorry in advance for the long post). I still strongly believe that the root cause of PFS has to do with epigenetic modulation. For such a persistent effect, there is not much from a molecular biology standpoint that could cause such and extended effect aside from epigenetic changes (e.g., methylation, acetylation) and genetic mutations. I highly highly doubt that the issue stems from actual changes to our DNA because there is nothing in the structure of finasteride that suggests it would have DNA mutagenic properties. There are also a dozen more reasons which I can get into, but basically, for those worried about the condition being passed on to children (I saw someone post about it) I wouldn’t worry about it.

One thing Awor mentioned to me got me thinking a lot. As I mentioned in my last post, I was a prostate cancer researcher in my previous life and awor asked me about a condition called hormone refractory prostate cancer. Normally, for advanced stage prostate cancer, cells still require androgen to grow. Thus in this advanced stage, patients are treated with “hormone therapy” which basically blocks androgen production and simultaneously blocks residual androgen from binding to androgen receptors (ARs) in the cancerous cells, thereby shutting down the growth signal. Eventually, in most patients their cancer comes back as hormone refractory prostate cancer. In this case, cells no longer require androgen to grow and hormone therapy (HT) fails to block growth. Interestingly, the anti-androgens in the HT appear to stimulate growth and adding androgen in some cases can even inhibit growth (in vitro). Basically the androgen response is reversed. This has been covered many times in the forum so apologies for ppl who already know this.

The cause of this insensitivity to androgen therapy is correlated with many factors including overexpression of AR and epigenetic changes. The point that Awor brought up is that the situation is analogous to PFS because when individuals with PFS administer testosterone, they feel nothing or even feel worse. What I’ve read again and again on the forum is that increasing androgen levels really doesn’t help. Again, this points to the issue being one of androgen responsiveness.

So thus far I haven’t said anything new. But, what I was thinking about was there is a good amount of research being done in the prostate cancer field on restoring sensitivity to hormone therapy. It would be feasible that a treatment which re-sensitizes prostate cancer cells to the inhibitory effects of androgen blocking therapy, would also cause cells to grow in response to androgen. And by correlation, we could hypothesize that it would also cause PFS sufferers to become sensitized to androgen again.

I did some searching on this topic to see whether anyone had looked into this, and I came across this article:

Restoring chemotherapy and hormone therapy sensitivity by parthenolide in a xenograft hormone refractory prostate cancer model
ncbi.nlm.nih.gov/pubmed/16921510

Basically parthenolide co-administration allowed androgen-resistant cells to once again be inhibited by hormone therapy. Now the interesting thing is that parthenolide is an HDAC inhibitor/epigenetic modulator ncbi.nlm.nih.gov/pmc/articles/PMC2672871/ which fits nicely into the hypothesis. It is also the active ingredient in an herb called Feverfew. Before any you go trying feverfew, remember that this experiment was basically done in petri dishes with pure parthenolide and there is even a chance it could have the opposite effect in vivo. Most compounds, even if available in dietary supplements get broken down during digestion in the liver so that there is only minimal active ingredient in the bloodstream. Not to mention that we don’t even know how much parthenolide, if any, is available in most feverfew dietary supplements since it is not regulated. It is more likely that you would not be able to get levels high enough in the bloodstream to feel an effect. However, if there were a well characterized FDA approved version available, I would really be interested in seeing if administering parthenolide would re-sensitize individuals w PFS to androgen. Since PFS often causes low levels of androgen, you would probably need co-administer with a testosterone gel.

Another interesting compound is panobinostat, which is currently being tested in humans at NYU:

“Using what is known about androgen receptors and their effects on prostate cancer growth, our researchers are designing clinical trials of new drugs with the potential to inhibit androgen receptor activity and restore the sensitivity of prostate cancer cells to hormonal therapies. One example is panobinostat (LBH589), an inhibitor of an enzyme called deacetylase. Panobinostat causes genetic changes in the androgen receptor, and laboratory studies have shown that it makes prostate cancer cells sensitive to the hormonal therapy bicalutamide. Our investigators are leading research evaluating panobinostat plus bicalutamide in men with castration-resistant prostate cancer.”

cancer.med.nyu.edu/researchers/r … r-research

Surprise surprise, it is also an HDAC inhibitor.

Okay, so now we’ve established there is some evidence for parthenolide (albeit pre-clinical and in a pretty mediocre journal) that one can restore sensitivity to androgen. And at least some confirmation of this rationale by evidence of an ongoing trial with panobinostat. Where else can we look for potential leads for restoring sensitivity to androgens?

I’m glad you asked. One interesting aspect of androgen receptor, is that it is part of a larger family of receptors called “nuclear receptors”. (en.wikipedia.org/wiki/Nuclear_receptor) These receptors reside in the cytoplasm of cells and bind to circulating hormones. Each type is specific for a certain type of hormone and once it binds, it undergoes a structural change and translocates to the nucleus, binds to specific DNA sequences, and begins transcribing (activating) genes which are involved in a variety of functions. In the case of AR, this includes genes responsible for sexual development and secondary sex characteristics. The interesting part is that the other hormone receptors share a good deal of structure with AR, utilize similar signaling intermediates, and are also regulated by similar epigenetic mechanisms. Even more interesting is that in some cases, these related hormone pathways can also become faulty and unresponsive to their respective hormones.

The closest example of this is estrogen receptor which is involved in breast cancer. Similar to prostate cancer, advanced breast cancer is also treated by blocking hormone activity. And again, eventually breast cancer cells become unresponsive to this therapy. Some searching led me to this article:

Valproic acid restores ER alpha and antiestrogen sensitivity to ER alpha-negative breast cancer cells.
ncbi.nlm.nih.gov/pubmed?term=19772891
Again, here is another example of an HDAC inhibitor which restores sensitivity to a hormone, albeit estrogen. Some of you may be familiar with Valproic acid as “Depakote” or a treatment for epilepsy and bipolar disorder. Looking at the side effects however, it looks like it is associated with reduced libido so it’s probably not a cure for PFS. Although, if anyone is using the compound I would be interested to hear what the effects are in people with PFS. I would also be interested to see if it increased sensitivity to testosterone at lower (or even higher) doses.

One last example I wanted to put out there is a condition seen with the glucocorticoid pathway (another hormone receptor pathway) in patients with COPD. COPD is a respiratory illness treated with corticosteroids which activate the GC pathway and reduce inflammation. At a certain point the body stops responding and loses sensitivity to glucocorticoids. A paper published last year showed that sulforaphane, a compound present in broccoli at low doses, was able to directly activate HDAC2 restoring sensitivity.

Denitrosylation of HDAC2 by targeting Nrf2 restores glucocorticosteroid sensitivity in macrophages from COPD patients
ncbi.nlm.nih.gov/pmc/articles/PMC3204828/

Although this study was a small one, sulforaphane is currently in Phase 2 development and it will be interesting to see how things pan out:
clinicaltrials.gov/ct2/show/NCT0 … ane&rank=3

Again with this product, the problem is that there is no pharmaceutical grade formulation, but it would nonetheless be interesting to see whether it is able to restore androgen sensitivity in a similar manner.

In conclusion, whether or not the above treatments work for PFS, I believe there is very strong rationale that PFS can be treated with epigenetic modulation. We have already seen three examples and I haven’t even begun to look at the other more than 20 hormone pathways. Furthermore, work on HDAC inhibitors is in its infancy and there are products coming out every year targeting new HDACs not to mention those targeting DNA methylation. This is good because you may need a combinatorial approach to targeting HDACs, activating some and repressing others, to effectively treat PFS. There are a daunting number of avenues to explore yes, but there are very promising leads with strong rationale.

Lastly, I just wanted to add a personal aside on this forum. I feel like all the members of this forum have been through a lot. Many feel jaded and hopeless, and have every reason to be. I know that it’s not much but as a formally trained molecular biologist, I am hugely optimistic. I see a mind-boggingly huge number of avenues to explore and possibilities for cure. I hope you got that sense even from just my brief research on hormone receptors which I presented here, that we are by no means at a dead-end in fighting this condition. In fact, with the establishment of the PFS Foundation and the new genomic tools being created everyday, we are just at the beginning. My colleagues in the biotech industry are in agreement that we are just at the early-stage of the genomic revolution. With personalized gene sequencing and gene expression analysis tools becoming cheaper and cheaper, over the next 10 years you will be seeing an advance in the understanding of disease that mirrors the impact the internet had on our lives. With these tools, and once we understand the molecular mechanism behind the disease, it’s really not a matter of how but when.

Thanks for reading this far and if anyone has had any experience with the above compounds, or have anything to add, I would love to hear it.

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Some very capable people have actually managed to get AR expression tested in a group of PFS sufferers.

Something was found but nothing that could explain the problem. viewtopic.php?f=33&t=6581 Should be published any day now.

And if its not the receptor (and post-translation studies are planned) arent these weak analogies about prostate cancer even weaker?

You can quote the exact post so you the reader knows which post you’re referring to. viewtopic.php?f=33&t=5282&start=160#p44242

I have no idea where you’re getting the “any day now” information. Don’t make shit up.

Wow,

this is a very good post.

Oscar, why are you so negative? You don´t know where the problem is, either…

We should get closer to the point where we can try such substances. Is this stuff used in treatment of prostata cancer?

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NYscientist
Thank your for your post.

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Great post!

Uplifting and positive… a nice change of pace around here.

I will be eagerly following this thread for more information.

Thank you!

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viewtopic.php?f=33&t=6581 “Publication: Q3/Q4 time frame” ??

Was I negative? This isnt a Disney cartoon, I dont know what you expect. If one claims to know the cause of ‘PFS’ be prepared to defend that assertion.

If androgen deprevation caused androgens to stop working prostate cancer would have been cured long ago. There can be methylation of the AR due to aberrant changes in some advanced cancer - but it now seems that hasnt happened to us. (also see; viewtopic.php?f=27&t=6968)

So how is Prostate cancer similar to our situation?

I brought up the use of Sulfpraphane a while back in the thread below…
viewtopic.php?f=9&t=6437&hilit=sulforaphane#p54738

“Sulforaphane” has been found to work both as a HDAC inhibitor and also a promoter of de-methylization."

=========

Every tablet contains 1,000mcg of Sulforaphane. Any clue as to what the starting point would be?
physicianformulas.com/store/ … roduct=267

Oscar, thanks for the feedback and I agree with some of your points. Also want to clarify again that I am in no way claiming to know the cause of PFS. In the absence of genomic data, we can only hypothesize and draw correlations with currently available research.

My thoughts, in the absence of data, is that androgen receptor (AR) underexpression (or overexpression) is not a route cause of PFS. If that were the case we would expect that adding supraphysiological doses of androgen would be able to compensate for underexpression of AR. You also bring up post-translational modifications. It is true that certain direct modifications to the AR receptor (e.g. phosphorylation) may alter the AR structure so it binds androgen less avidly, or impacts its ability to interact with key co-activators. One of the reasons I feel that this is also not the case in PFS is that after reading lots of member stories, it seems like in many cases people are exhibiting responsiveness to androgen in some areas but not others. For example, hairloss continues while libido is virtually unaffected. This implies that in PFS, we are experiencing selective inhibition of a subset of androgen responsive genes. If it was a general problem with our AR (structure, levels of expression, post-translational modification), we would see a global decrease in all androgen responsive sex characteristics similar to true AIS.

Now there are only a few ways that selective gene modulation can occur, and one major way is through epigenetic mechanisms such as acetylation. In fact, when you treat patients with HDAC inhibitors, you see that each inhibitor of a specific HDAC affects a different subset of genes, upregulating some genes while down-regulating others.

True, we have not seen recorded instances of males becoming insensitive to androgen in the exact way we theorize for PFS. But, this does occur frequently and is quite well documented in another hormone receptor pathway, the glucocorticoid pathway. As mentioned earlier, over-activating this pathway with corticosteroid (the equivalent of androgen in this pathway) causes extreme and irreversible de-sensitization to corticosteroid. It is not unreasonable to think that such fail safe mechanisms exist for other pathways in the hormone receptor family. Furthermore, (it’s one study so we have to take it with a grain of salt) it was demonstrated that an HDAC inhibitor (sulforaphane) was able to restore some sensitivity to the ligand.

I’m admittedly pretty unfamliar with the whole 3adiolG and 3a-HSD area so can’t comment there but if you have any links to good review articles please PM them to me.

Reading the comments here make me realize just how important it is that we support research that helps us better understand the molecular basis of PFS in any way possible whether it be spreading the word about PFS, donating, or participating in studies. The studies that Awor is helping with, particularly the full gene expression analysis, epigenetics, and proteomics studies are the key to the cure. In fact, it is quite possible the cure is already out there but we just don’t know which drug(s) to use until we have the target.

Moonman1, I’m also thinking about trying sulforaphane out but it’s difficult to tell what the appropriate dose would be because we don’t know how much is getting broken down in gut and how much (if any) makes it into the bloodstream, what the half-life in the bloodstream is, and how consistent the dose is per tablet (dietary supplements may have widely ranging levels of active ingredients by tablet). If you want to test it yourself please be careful. The way it is done in clinical trials, is that they start at dose they feel is safe (maybe two tablets), take it for a few weeks consistently, and if it is well tolerated, increase the dosage by ~30-50% each time (e.g., 2000, 3000, 4500, etc…). In the meantime, you would keep a very close watch on side effects, particularly those common with HDAC inhibitors: nausea, vomiting, severe fatigue, infections, loss of appetite, skin reactions, and diarrhea to name a few. Once you find the “maximum tolerated dose”, you would scale it back to the previous dose. Because PFS is associated with low androgen levels, I would think it would increase the likelihood of seeing something if one added a testosterone gel to the regimen after selecting the maximum dose.

Okay, I just did a quick google search and came up with this article which discusses sulforaphane levels in humans.
mct.aacrjournals.org/content/6/3/1013.full

It looks like it gets absorbed into the blood stream but the problem is it has a really short half-life of 1.8h (i.e., every 1.8 hrs the blood concentration drops by half as it gets broken down). I’m not a pharmacologist, but it seems like any experiment with sulforaphane, you will have to dose it at least every 4 hours or so if you really want to see if it has an effect. That’s a lot of pills in a day! In general, I think with a lot of these dietary supplements people are trying out, they are not going to see anything because they are already starting out with too low a dose, or the drug half-life is too short.

As an aside, has anyone on the forum looked into manufacturers of dietary supplements to see which are providing the absolutely highest quality and most regulated dietary supplements? I wouldn’t trust the stuff that you find at GNC.

Also, has anyone put together a list of commercially available HDAC inhibitors? Whether it be dietary supplements or FDA-approved ones. Over the next few months, I’m going to be testing some of these alone and in various combinations to see if they have an impact on androgen-response.

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Hey nyscientist

Thanks for your positive posts, it’s a rare thing around here.

You’ve went into a great deal of detail here, is there anything we can try or do practically? Most of us here are willing to try almost anything.

You mentioned diet briefly, can you elaborate on your thinking on this?

We should build a team to try all those medications and supplements. Don´t know if I am wright, but with hormone therapie you can stop cancer cells from growing and not kill them, or Oscar? So if there is a way to get cancer cells sensitive to hormones again, why this cannot work with us?

Oscar the more and more I read what you are writing the more I see, that you have no clue what you are talking about. There are scientists working on this and you with your basic knowlage, belive you know it always better. Some things you are saying are not totaly wrong, but it shows, that for you the basics and some important things are missing. You hijack every treat and try to attack awor in any way. I think nearly everybody here in this forum, has seen what you think of this. I dont see awor doing this and for me it realy looks like you spend more time with awors theory than with your owen. Go and prove what you are saying! You do nothing more than just attacking others, driving people crasy for me you are more and more just a troll!!! Who messes up people and try to make awarenes, how you claim it. We all are sitting in the same Boat and we all want a cure for this shit. Or dont you want? Or do you only want to slpit people make belive wars? What is your aim? You let it look awor is an enemy, I see many many things, he did for us! What have you done? You just attacking him everywhere in every post. You realy suck!

“I just take this: if androgen deprivation caused androgens to stop working Prostata cancer would have benn cured long ago”

What a BS. You take things and belive its all so simple, what is not true. I´m realy sick of reading your stupid and absolut unprofessional posts!

lifewasgood, sorry about confusion, when I say “dietary supplements” I am referring to various herbs and other nutritional products that are not regulated by the FDA as drugs. The FDA classifies these products as dietary supplements and they are regulated much less strictly. My question was what is the best source to buy these medicinal herbs and nutritional products. I would be interested in trying a few of these products like sulforaphane and feverfew but would want to ensure that we use the absolute best quality available since quality ranges dramatically.

I would also be in for trying a few of these products as a group (mostly the safer ones), some people may respond differently than others and it would raise the possibility of catching a positive signal. Similar to what I mentioned for sulforaphane, it would take a bit of research to think of the right protocol for each. Maybe we could try a new approach every few months. No guarantees that we would see anything, but at least would be a way to keep occupied until the clinical studies and data are available to really understand what’s going on. Is there a section of the forum where we can organize stuff like that?

have a look here: viewtopic.php?f=5&t=3901&start=460

Here is the Sulforaphane product that a lot of the research studies use. Its a bit expensive, but most likely the highest quality…

sigmaaldrich.com/catalog/pro … &region=US

Thanks for your contribution, NYscientist. It’s highly appreciated. It’s good to see some well trained and highly knowledgable members here. You should hook up with the research initiatives here.

I would be wary of using this one. It comes as an oil and not sure how well it would be tolerated. Correct me if I’m wrong, but it seems like the studies using this are all in vitro experiments, which is basically adding this to a petri dish of cells and seeing what effect it has. I didn’t see any papers where they were actually administering this formulation to patients which is very different. However, if there are examples, I would feel much more comfortable trying it.

It looks like when using in humans, people are using broccoli sprout extract or preparing their own formulations such as in the following study:

“Quantitative determination of dithiocarbamates in human plasma, serum, erythrocytes and urine: pharmacokinetics of broccoli sprout isothiocyanates in humans”
(You can find the paper if you search on Pubmed).

The next two weeks I will do some research to find the best source for broccoli sprout extract. I wouldn’t trust most dietary supplement manufacturers on face value as you don’t know what you are getting. Ideally the company would provide hard data on the sulforaphane content of their product. If any of you have some spare time and can do some research as well to find the best quality suppliers that would help. Once I find a good supplier, I will try to come up with a good protocol we all can try and make a new post. Moonman1 and others looking to try this out, it would increase the chances of seeing a signal if you are able to obtain androgel or some other topical testosterone formulation (androgel is like $400 if not covered by your insurance but you can obtain it from compounding pharmacies that make their own formulations for ~$50, I have the address of one if anyone is interested). My thoughts are, we may only be able to get suboptimal concentrations of sulforaphane in the bloodstream so boosting the testosterone signal may increase the prospects of finding something.

Brainbug, there are a lot of good leads on the link you posted for other HDAC inhibitors to try. I can try to put together some other protocols with those afterwards.

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Hey guys, this is another interesting and potentially exciting article I found on valproic acid (HDAC inhibitor) enhancing androgen responsiveness (albeit in vitro).

ncbi.nlm.nih.gov/pubmed/21862211

Valproic acid, also known as Depakote is an HDAC inhibitor used for bipolar disorder. Normally it is associated with a decrease in libido when used constantly, however the exciting thing that this paper found, is that if you pre-treat an androgen resistant prostate cell line for 2 days, and then treat with androgen (DHT), it increased androgen responsiveness (as measured by growth) by 70-120%! Now this is a study done in cells, but since Depakote is an approved prescription drug, it should be really easy for someone to try this out. Does anyone have means of (legally) obtaining some Depakote?

I’ve included the specific paragraph of interest below:

"…We thus investigated the effect of HDAC inhibitor on androgen-responsive cell growth. Androgen-independent C-81 cells were treated with 1 mM VPA for 48 h followed by 10 nM DHT in a steroid-reduced condition. The usage of 10 nM DHT is to determine if VPA pre-treatment can increase the common androgen-responsiveness of cell growth. In the presence of DHT for 48 h, the growth of VPA-pretreated C-81 cells increased by about 70% (column #4 vs. #3, Fig. 3B, left panel, p < 0.01), compared with only about 35% increase of control cells without VPA pre-treatment (column #2 vs. #1, Fig. 3B, left panel). Western blot results validated that the cPAcP 50 kDa mature form protein was elevated by VPA treatment (lane #3 vs. #1, Fig. 3B, right panel), which was decreased by subsequent DHT-treatment (lane #4 vs. #3, Fig. 3B, right panel), inversely correlating with cell growth stimulation [20], [27] and [28]. In parallel, cellular PSA level was greatly elevated by DHT in VPA-pretreated cells, about 1.5-fold of that in control cells without VPA-pretreatment (lane #4 vs. #2, Fig. 3B, right panel). In those same cells, AR expression level was not significantly changed after a total of 5 days treatments including 2-day by VPA and 3-day by DHT.

Due to the clinical importance of androgen sensitivity of PCa cells, we investigated whether VPA treatment could similarly increase the degree of androgen responsiveness in other androgen-independent PCa cells, including LNCaP C4-2 and MDA PCa2b-AI cells. As shown in Fig. 3C for LNCaP C4-2 cells and Fig. 3D for MDA PCa2b-AI cells, DHT alone could increase the basal cell growth by approximately 10% (column #1 vs. #2). Interestingly, DHT could greatly increase the growth of VPA-pretreated cells by about 70% and 120%, respectively, (column #4 vs. #3, Fig. 3C and D, left panels). We subsequently validated DHT effect by semi-quantifying PSA levels in those treated cells. Interestingly, PSA basal level was elevated in VPA alone-treated MDA PCa2b-AI cells in the absence of DHT (lane #3 vs. lane #1, Fig. 3D, right panel). Importantly, cellular PSA level was greatly elevated by DHT in VPA-pretreated cells, approximately 4- and 9-fold of that in control cells without VPA-pretreatment, respectively (lane #4 vs. #2, Fig. 3C and D, right panels). It should also be noted that due to the low expression level of AR protein in these two PCa cell lines, comparing with LNCaP C-81 cells, a prolonged hybridization with primary Ab to AR with longer exposure time periods was required. In summary, VPA pre-treatment enhances DHT effect on the increments of cell growth and PSA expression, which indicates that HDAC inhibitors can enhance androgen responsiveness of PCa cells."

NYscientist,

You state in your opening post that we shouldn’t worry about passing this to our children (those of us who can have them) because Fin should not have altered our DNA itself. You also reference that likely some epigenetic change has taken place that makes our problem persistent. Many others seem to agree with this and for now it seem to be one direction of the current research. Lately alot of other research has shown that epigenetic changes can not only be inheritable but can last for generations.

This is just one of many papers which references multigenerational epigenetic change. pnas.org/content/109/23/9143

Further, we’ve had at least one anchedotal report here on the board of PFS being passed on from father to son.

Would be interested to hear your thoughts.

God I hope not, I have a baby boy coming next month, so far all is normal.