).
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To Swooping and Chemical: thanks guys - you both seem to know the science cold. And again - FGF11 - appreciate your insights. This is a pretty high level thread. You could probably stitch enough together to make for a fancy review article in a journal (not that we care about that ).
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Also I don't remember using the word cure. I have a philosophical problem with that word.
So just to clear things up again, please don't say that I have said I have the cure. I've mentioned many times that I don't. So many times, that even, someone thought, I said it's ten years away.
I see some intelligent comments showing up. That's great people. Thanks.Leave a comment:
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Swooping,Why you think that secondary activation of AR is that important through non-androgen dependent pathways? If that were to be true then why do castrated people never develop AGA if they are
castrated before puberty? Secondly why does their balding stop when they are castrated when suffering from AGA? If secondary activation through non-androgen dependent pathways was a problem it would be expected that AR would still cause problems in these people, but it doesn’t. Their balding stops. Injecting these castrated people with testosterone causes their hair to fall out again. This has been all shown ages ago by Hamilton.
I think Cotsarelis puts this well;
So, yes AGA is androgen dependent. Remove androgens and AGA ceases to exist pretty much (the same could be said for AR).
If you remove androgens or AR then all downstream factors that were harmful before become irrelevant. After all balding stops when you remove androgens, so apparently these downstream factors that were harmful before suddenly are not damaging anymore in terms of progression of AGA.
So indeed that begs the question; why doesn’t the hair follicle revert to the original state in that case? Many people have problems with this fact. But here is exactly where the consensus of many researchers comes into place. It’s simply damage over time that leads to an altered cell state. I don’t know why many people seem to neglect these papers/evidence. It’s way more accepted for instance than the PGD2 hypothesis. Way more logical too, at least to me.
Take a piece of plastic and heat. Now the piece of plastic is the hair follicle and the heat in this case is the damage (Androgens and AR upstream signals known). Eventually the plastic will start to melt and adopts a new form. Now your remove the heat but does the plastic re-shape itself to it’s original form? I don’t think so.
If you are interested more into this literature I suggest you to start reading about this. Here are some to start;
I would just like to call AR activation due to androgens “stress”. This stress sets in motion extremely major important pathways that alter these cells. These are not factors like PGD2, TGF-b, DKK1. Nah, it gets more way more important than that and these studies clearly show these pathways. All the microRNA studies also reflect upon these pathways.
Now one might ask the question; But hey we remove all androgens so these major important pathways don’t get activated anymore, so why the hell doesn’t my hair follicle reverse to it’s original state? Well wouldn’t that be inefficient? It's just how it works.
If we look for example on P53 only just as a pure example (don't look at the tumor suppression although this is also a function of P53);
Get it? P53 for instance (which is over expressed in scalp as you can see in the study) can decide that damage is to big and it can cause apoptosis or permanent cell cycle arrest and create a very tough "lock" if that makes sense to you. Now it might not be P53 which is causing this but you get the point. It's perhaps some sort of efficient lock, a lock that got activated due to the stress on the cells. Or apoptosis or a combination of both. I'm not saying that P53 is implicated it just acts as an example, but evidence has been shown in papers that it is probably implicated. Go have a read for fun on the function of P53 overall.
Anyway there could be said much more about this. Everything falls into place and makes sense if you read those studies and understand what these pathways do and why they act like that. I think clearly the "broad" picture is becoming clear.
Anyway didn't mean to hijack this topic. Answers your question why full AR silencing won't act as a cure or reverse hair miniaturization in to great extent. Sure some people will have some regrowth. But guess what some people also have some regrowth with finasteride/dutasteride/RU58841. In this case I guess (some) damage of the cells was still repairable. But acting as a cure? Nah, doubt it very much. Highly unlikely.
Burden of proof is on OP anyway, cause he is the one making very bold claims.
I know about stress and senescence. I've read those papers.
I get them. I can make something up here easily, I don't want to cherry pick or skew the facts when coming to theories. It's easy to pretend you talk science, but no studies so far have been done on what I wrote. The only study so far is what I've done, with my very limited resources. It will be interesting to investigate Nuclear translocation of AR of HFs, for example, in Castrated-Mice in presence and absence of selective cytokine signals to see its effect on HF cycle. Or to see the effect of cytokines, in HFs, on migration and secretion of Growth Factors. Or analyze if cytokine treatment of transplanted Human HF, will cause AR nuclear localization in the back of SCID mice or not. So many studies that can be done.
Or I can say for example:
1) Yes, they don't go bald (Castrated Males) Why?
Since androgen as an initial signal is needed to get cells activated to produce cytokines in the first place.
When androgen is totally absent cytokines, are not made in the HFs, and the eventual add up of cytokines therefore never happen.
Now, imagine, when cytokines (or some other factors) levels are low, the activation of AR is majorly androgen dependent, while they start to add up, it become androgen independent, lessening and lessening the need for androgen-dependent signalling.
2) It's not only about cytokines, I'm talking the big picture here. AR, will lock other transcription factors such as FOXa2 and Beta-catenin, to have their normal function in the nucleus. So, It's knockdown will cause secretion a set of growth factors, such as WNTs, and activating epithelial stem cells in a co-ordinate manner to divide.
3) Or I can stress is reversible (my M.Sc. was on stress signals and ERAD) and through some papers for it (but you seem to have a biological background so you know what I say)
The truth though is that we don't know.Leave a comment:
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So why are you here? Just to piss on people?Do you all always go take a piss when Spencer says: "Take everything you read on the forums with a grain of salt"?
No way I'm reading all that crap, some of us have lives. I will trust real world scientists over forum mad scientists any day.
Have fun making another of these 100 pages thread who will be buried and forgotten in a few years. Joining the hall of fame made of threads like these:
So much energy spent and what did we gain from these threads? Jack.Leave a comment:
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I'd like to expand on what swooping just mentioned, since I've taken the time to actually read those studies relating to P53, p21, p16 what exactly their roles are in tumor suppression.If you are interested more into this literature I suggest you to start reading about this. Here are some to start;
I would just like to call AR activation due to androgens “stress”. This stress sets in motion extremely major important pathways that alter these cells. These are not factors like PGD2, TGF-b, DKK1. Nah, it gets more way more important than that and these studies clearly show these pathways. All the microRNA studies also reflect upon these pathways.
Now one might ask the question; But hey we remove all androgens so these major important pathways don’t get activated anymore, so why the hell doesn’t my hair follicle reverse to it’s original state? Well wouldn’t that be inefficient? It's just how it works.
If we look for example on P53 only just as a pure example (don't look at the tumor suppression although this is also a function of P53);
Get it? P53 for instance (which is over expressed in scalp as you can see in the study) can decide that damage is to big and it can cause apoptosis or permanent cell cycle arrest and create a very tough "lock" if that makes sense to you. Now it might not be P53 which is causing this but you get the point. It's perhaps some sort of efficient lock, a lock that got activated due to the stress on the cells. Or apoptosis or a combination of both. I'm not saying that P53 is implicated it just acts as an example, but evidence has been shown in papers that it is probably implicated. Go have a read for fun on the function of P53 overall.
Anyway there could be said much more about this. Everything falls into place and makes sense if you read those studies and understand what these pathways do and why they act like that. I think clearly the "broad" picture is becoming clear.
It is a known fact Androgens cause DNA damage, this is in part due to evolutionary adaptations and stupid mutations. How and why is the crucial topic of interest which people love avoiding.
This study shows that Cyclin d1, a cell cycle progression protein is necessary to mediate DHT dependent DNA damage. Testosterone mediates most of its androgenic effects via autorine/paracrine conversion to DHT which binds to nuclear or neighbouring cell surface AR receptors.
Cells must proliferate and regenerate continuously, therefore it is inevitable that they will accumulate DNA damage over time, and moreso when you add in reactive oxygen species. Androgens increase IGF-R, there are many studies outlining this, but IGF-1 is a known cancer promoting agent. IGF-1 causes enhanced cell hypertrophy and proliferation, which translates to more Cyclin d1 => more DNA damage. This is why the body has developed counter measures to protect against this. The tumor suppressor family of p53/p21/p16 all exert their effects by reducing proliferation of cells and achieving premature senescence. If the cells continue to proliferate with DNA damage, they will end up becoming cancerous. Our ancestors that developed these feedback loops survived, explaining why we have them in the first place.
What do these p-family genes do?
p16:
So it inhibits the proliferation of cells and promotes cellular senecense (cells have a certain number of replication cycles determined by telomerase - which is why we have stem cells, and even they can achieve senescence which is the secret to anti-aging but thats another post)p16 is a cyclin-dependent kinase (CDK) inhibitor that slows down the cell cycle by prohibiting progression from G1 phase to S phase. Normally, CDK4/6 binds cyclin D and forms an active protein complex that phosphorylates retinoblastoma protein (pRB). Once phosphorylated, pRB disassociates from the transcription factor E2F1, liberating E2F1 from its cytoplasm bound state allowing it to enter the nucleus. Once in the nucleus, E2F1 promotes the transcription of target genes that are essential for transition from G1 to S phase.[9][10]
p16 acts as a tumor suppressor by binding to CDK4/6 and preventing its interaction with cyclin D. This interaction ultimately inhibits the downstream activities of transcription factors, such as E2F1, and arrests cell proliferation.[10] This pathway connects the processes of tumor oncogenesis and senescence, fixing them on opposite ends of a spectrum. On one end, the hypermethylation, mutation, or deletion of p16 leads to downregulation of the gene and can lead to cancer through the dysregulation of cell cycle progression. Conversely, activation of p16 through the ROS pathway, DNA damage, or senescence leads to the buildup of p16 in tissues and is implicated in aging of cells.[9]
p21:
once again similar to p16 but has additional effects on DNA damage repair.p21 is a potent cyclin-dependent kinase inhibitor (CKI). The p21 (CIP1/WAF1) protein binds to and inhibits the activity of cyclin-CDK2, -CDK1, and -CDK4/6 complexes, and thus functions as a regulator of cell cycle progression at G1 and S phase.[5] In addition to growth arrest, p21 can mediate cellular senescence. One of the ways it was discovered was as a senescent cell-derived inhibitor.
p53:
p53 has a far more diverse role. Not only does p53 inhibit cell cycle progression, it attempts to repair the damage. A good thing right? it would be if thats all did. P53 is good for the prostate because it tries to prevents DHT driven prostate cancer via WNT. It inhibits the canonical WNT signalling pathway (aberrant wnt signalling leads to cancer ), however, stupidly, the prostate is incredibly similar to the scalp - I've covered this in more detail in my other thread.p53 has many mechanisms of anticancer function and plays a role in apoptosis, genomic stability, and inhibition of angiogenesis. In its anti-cancer role, p53 works through several mechanisms:
It can activate DNA repair proteins when DNA has sustained damage. Thus, it may be an important factor in aging.[30]
It can arrest growth by holding the cell cycle at the G1/S regulation point on DNA damage recognition (if it holds the cell here for long enough, the DNA repair proteins will have time to fix the damage and the cell will be allowed to continue the cell cycle).
It can initiate apoptosis (i.e., programmed cell death) if DNA damage proves to be irreparable.
p53 induces DKK1 to inhibit WNT signalling and WNT binding to LRP6:
p53 Activates MicroRNA-34 to Inhibit Wnt Signalingp53 is a tumor suppressor and loss of p53 function accelerates mammary tumorigenesis by Wnt. In this study, we found that Dkk-1 is induced by wild-type p53 but not mutant p53(R249S). In addition, DNA damage upregulates Dkk-1 in cell lines that harbor an endogenous wild-type p53 gene but not in cell lines that are p53-null or harbor an endogenous mutant p53 gene. We also found a potential p53 responsive element located approximately 2100 nucleotides upstream of the Dkk-1 transcription start site and we show that p53 binds specifically to this element both in vitro and in vivo. Furthermore, we have established several cell lines derived from H1299 lung carcinoma and U118 glioma cells that inducibly express Dkk-1 under a tetracycline-regulated promoter. We found that Dkk-1 has no effect on proliferation of cells that are not transformed by Wnt. Taken together, these results suggest that Dkk-1 may mediate p53 tumor suppression by antagonizing the Wnt signaling pathway.
now why the f*** would you inhibit WNTs in hair follicles? We're not going to frickin die from excessive hair growth. I still dont understand why the body has these mechanisms, its just blindly evolving using whatever works in the short term, but that's the beauty of evolution i guess.
so to conclude, yes androgens cause senescence and DNA damage, but they work through pathways that can be targeted/exploited with different and feasible strategies than just silencing the very broad Androgen receptor.
Personally I think the androgen receptor - if inhibited, can allow other treatments to work far more effectively. FGF11s research has promise but I'd like some more supporting evidence both in terms of theory and practical use.Leave a comment:
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Do you all always go take a piss when Spencer says: "Take everything you read on the forums with a grain of salt"?
No way I'm reading all that crap, some of us have lives. I will trust real world scientists over forum mad scientists any day.
Have fun making another of these 100 pages thread who will be buried and forgotten in a few years. Joining the hall of fame made of threads like these:
So much energy spent and what did we gain from these threads? Jack.Leave a comment:
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Why you think that secondary activation of AR is that important through non-androgen dependent pathways? If that were to be true then why do castrated people never develop AGA if they are
castrated before puberty? Secondly why does their balding stop when they are castrated when suffering from AGA? If secondary activation through non-androgen dependent pathways was a problem it would be expected that AR would still cause problems in these people, but it doesn’t. Their balding stops. Injecting these castrated people with testosterone causes their hair to fall out again. This has been all shown ages ago by Hamilton.
I think Cotsarelis puts this well;
So, yes AGA is androgen dependent. Remove androgens and AGA ceases to exist pretty much (the same could be said for AR).We know a lot about AGA from studies a long time ago, in the ’50s, by Dr. James Hamilton. He was the first one to coin the term androgenetic alopecia. He noticed that men who had been castrated before puberty never went bald, and realized that baldness was androgen-dependent. He did some experiments where he gave men who had been castrated testosterone; the ones who had a family history of baldness started to go bald, so that was how he identified the genetic component. He originally called it androchronogenetic alopecia because you needed androgens, time, and genetics. We’ve known it’s androgen-dependent since then.
If you remove androgens or AR then all downstream factors that were harmful before become irrelevant. After all balding stops when you remove androgens, so apparently these downstream factors that were harmful before suddenly are not damaging anymore in terms of progression of AGA.
So indeed that begs the question; why doesn’t the hair follicle revert to the original state in that case? Many people have problems with this fact. But here is exactly where the consensus of many researchers comes into place. It’s simply damage over time that leads to an altered cell state. I don’t know why many people seem to neglect these papers/evidence. It’s way more accepted for instance than the PGD2 hypothesis. Way more logical too, at least to me.
Take a piece of plastic and heat. Now the piece of plastic is the hair follicle and the heat in this case is the damage (Androgens and AR upstream signals known). Eventually the plastic will start to melt and adopts a new form. Now your remove the heat but does the plastic re-shape itself to it’s original form? I don’t think so.
If you are interested more into this literature I suggest you to start reading about this. Here are some to start;
I would just like to call AR activation due to androgens “stress”. This stress sets in motion extremely major important pathways that alter these cells. These are not factors like PGD2, TGF-b, DKK1. Nah, it gets more way more important than that and these studies clearly show these pathways. All the microRNA studies also reflect upon these pathways.
Now one might ask the question; But hey we remove all androgens so these major important pathways don’t get activated anymore, so why the hell doesn’t my hair follicle reverse to it’s original state? Well wouldn’t that be inefficient? It's just how it works.“a process that is thought to reflect irreversible cell growth arrest in the progression of AGA.”
If we look for example on P53 only just as a pure example (don't look at the tumor suppression although this is also a function of P53);
Get it? P53 for instance (which is over expressed in scalp as you can see in the study) can decide that damage is to big and it can cause apoptosis or permanent cell cycle arrest and create a very tough "lock" if that makes sense to you. Now it might not be P53 which is causing this but you get the point. It's perhaps some sort of efficient lock, a lock that got activated due to the stress on the cells. Or apoptosis or a combination of both. I'm not saying that P53 is implicated it just acts as an example, but evidence has been shown in papers that it is probably implicated. Go have a read for fun on the function of P53 overall.
Anyway there could be said much more about this. Everything falls into place and makes sense if you read those studies and understand what these pathways do and why they act like that. I think clearly the "broad" picture is becoming clear.
Anyway didn't mean to hijack this topic. Answers your question why full AR silencing won't act as a cure or reverse hair miniaturization in to great extent. Sure some people will have some regrowth. But guess what some people also have some regrowth with finasteride/dutasteride/RU58841. In this case I guess (some) damage of the cells was still repairable. But acting as a cure? Nah, doubt it very much. Highly unlikely.
Burden of proof is on OP anyway, cause he is the one making very bold claims.Last edited by Swooping; 11-30-2015, 11:52 AM.Leave a comment:
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Summary of FGF11's Findings
You need to de-activate the androgen recepetor (AR), because it is activated by both Hormones (DHT) and cytokines. Thats why finasteride is not a total solution, it acts on the dht side of the equation but not the cytokine problem.
Thanks so much FGF11Leave a comment:
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Thanks so much for your contributions FGF11 - they are really intriguing.
Daily injections over your entire scalp is obviously not ideal - but it may not be necessary. Earlier this year a company called Prometheon Pharma (http://www.prometheonpharma.com/) was touting a trans-dermal delivery vehicle for large molecules (as big as insulin) and tried to use it with IGF-1 to regrow hair. Not surprisingly, it failed to regrow hair because IGF-1 is insufficient to combat AGA.
But their trans-dermal delivery vehicle could be ideal for oligonucleotides - effectively converting your therapy into a topical cream that could be applied daily.
Thoughts? Thanks again for your insights.Leave a comment:
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Interesting. Yeah - I guess we don't know what those variants are tagging - but it definitely involves the AR. The real culprit is probably a non-coding SNP that impacts AR levels. People who don't go bald probably have much lower AR levels in their follicles - and hence no matter how much DHT is circulating, it's not sufficient to trigger the AR positive feedback loop that triggers balding - there's just not enough AR to reach the necessary threshold. Interesting about EDA2R - and again - it looks like it interacts with AR - ? also involved in its upregulation or increased activity levels.
It all still fits with his theory - reduce AR in the scalp and you will solve balding. Very encouraging that it looks like other groups have already worked on developing oligonucleotides to reduce its expression - that's exactly what OP was doing - it definitely seems feasible.Leave a comment:
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Agree - proof is critical.
But how can you be certain that silencing of AR won't reverse miniaturization? As highlighted - this is not the same as complete androgen deprivation (like what happens with trans-genders). As we know, trans-genders with pre-existing baldness do not regrow most of their - but as he indicated, this may be secondary to continued activation of the AR through non-androgen dependent pathways.
I don't think we have any evidence to indicate that silencing of AR will not regrow hair - has this ever been done/tried before? If his theory is correct, the AR is the final common pathway - everything converges on the AR to trigger and maintain baldness. If you reduce/eliminate AR, follicles should be released and able to grow again - regardless of how much PGD2, TGF-beta, etc. that you have floating around your scalp.Leave a comment:
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Hi sorry for the noob question- does that mean, if im reading it right- that ablating the AR receptor in the hair follicles on the hairless scalp will resolve hair loss and regrow hair?Leave a comment:
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To make more sense out of antisense oligos:
#1 Most of topical solutions act through HFs, what it means is that many of the topical solutions that deliver oligonucleotides pass epithelium into dermis, through the hair shaft (maybe). They concentrate there. Now, most of the studies done in hairless mice with oligo topical solutions (for silencing other genes) do not work (usual topical solutions I mean). However, the same studies work in mice with HFs. Therefore, if the area, is not covered with visible large HFs, the efficiency of Oligo should be extremely low.
#2) The best scientific experiment would be injection (PLEASE DO NOT TRY THIS). But if I were to suggest something, I would say a microneedle semi-auto tattoo gun (they are dermal guns), would probably be the best away to efficiently and vastly introduce oligo every three - four days, fast for a large area. However, it will be inefficient, and more expensive.
3) You should remember, maybe, the physical disruptions, introduced while adding the oligos, were MAYBE HELPING in disrupting the fibrosis (whether it is there or not). Injecting intradermally, makes a gap between dermis and epithelia, for about 30 minutes, every time. By pushing epithelia upward, this may (theoretically) be the reason why the topical would not work.
4) Yes, there are a couple of patents regarding silencing AR that may or may not (I'm not a lawyer) be able to lock the technology. None of them, however, to my understanding, have really tried this, even for ONCE. They did only file the patent, for when such a day would come. Even there are one or two studies trying to silence AR. Those studies are not good studies.
5) This is NOT going to be the ultimate treatment (Please don't use the word cure referring to hair loss - it's an aesthetic condition). This however, will be, extremely more effective treatment with oligo's, and it may actually be very SAFE.
6) Triple repeats in AR do not only correlate with AGA but they also correlate with rheumatoid arthritis (RT). Pointing toward a role for AR in AGA and RT.
I appreciate when you share intelligent comments. Please contribute intelligently, otherwise, don't.Leave a comment:
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Thanks for your effort.
Inactivation/silencing of AR will most definitely not reverse miniaturization though. I'm extremely sure of this so I automatically put your story into question.
Proof is what I want to see.Leave a comment:
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First of all I want to really thank you FGF11 for your experiment. However some photos as a proof would be gently welcomed (I belive in this theory even without a proof though)
I am posting some studies connected to silencing mutated AR. Probably there is a way to create topical solution with minimal side effects and if your claims are true - that would be a next generation ULTIMATE CURE.
So please FGF11 show us the proof, after that we should try to make this gene therapy happen.
Silencing the androgen receptor: New skills for antiandrogen oligonucleotide skin and hair therapy
Control of androgen receptor expression in human keratinocytes and in a reconstituted human epidermis model with selective antisense oligonucleotidesAntiandrogen therapeutic oligonucleotides targeting the downregulation of the ARexpression is advantageous because both will be possible to eliminate the only way for androgens to act and simultaneously this strategy allows the medication to be topically administrated.
Association of locally increased androgen activity and skin disorders is obvious in acne and androgenetic alopecia in males. In addition, testosterone was unexpectedly found to perturb the epidermal barrier. Blockade of androgen action via androgen receptor (AR) antagonism accelerates wound healing in aged individuals. Androgen activity on skin can classically be inhibited by systemic administration of compounds, which have strong affinity for AR and antagonize androgen binding to AR molecules. In this study we applied a new technology to realize the same purpose: We tested the activity of antisense oligonucleotides against the AR in primary human foreskin keratinocytes, human non-foreskin keratinocytes from young (30 y) and older (60 y) female donors, and reconstituted human epidermis (SkinEthic model). Reconstituted human epidermis is similar to in vivo human epidermis and features a functional permeability barrier. To transfer the antisense oligonucleotides into human keratinocytes an optimum liposome-mediated transfection system with Poly-L-ornithine (12 μg/ml) over 4 h was used. The transfection efficiency was assessed using FITC-labeled (ACTG)5 random oligonucleotides, which were localized in cytoplasmic structures of the keratinocytes. AR expression on the protein level was investigated by Western blotting. Transient transfection of foreskin keratinocytes with phosphorothioate antisense oligonucleotides (PTO) revealed a reduction of AR expression (≈25%) compared to native keratinocytes after 14 h recovery time. The AR knock down in epidermal keratinocytes of the compared women was stronger in the older, more differentiated keratinocytes. After 24 h, AR expression level have returned back to the level of non-transfected cells. The effect could be reestablished by repetition of transfection. PTO and 2?O-methylribosyl (MRO) antisense oligonucleotides decreased AR expression at levels varying between 46% and 70% in the air-lifted reconstituted human epidermis after 18 h recovery time. The successful inhibition of AR expression in human keratinocytes and reconstituted human epidermis is the first step to develop topically efficient compounds with oligonucleotidesLeave a comment:
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