Rapamycin treatment increases survival, autophagy biomarkers and expression of the anti-aging klotho protein in elderly mice

#1

Abstract

Previous investigations have demonstrated that treatment of animals with rapamycin increases levels of autophagy, which is a process by which cells degrade intracellular detritus, thus suppressing the emergence of senescent cells, whose pro-inflammatory properties, are primary drivers of age-associated physical decline. A hypothesis is tested here that rapamycin treatment of mice approaching the end of their normal lifespan exhibits increased survival, enhanced expression of autophagic proteins; and klotho protein-a biomarker of aging that affects whole organism senescence, and systemic suppression of inflammatory mediator production. Test groups of 24-month-old C57BL mice were injected intraperitoneally with either 1.5 mg/kg/week rapamycin or vehicle. All mice administered rapamycin survived the 12-week course, whereas 43% of the controls died. Relative to controls, rapamycin-treated mice experienced minor but significant weight loss; moreover, nonsignificant trends toward decreased levels of leptin, IL-6, IL-1β, TNF-α, IL-1α, and IGF-1, along with slight elevations in VEGF, MCP-1 were observed in the blood serum of rapamycin-treated mice. Rapamycin-treated mice exhibited significantly enhanced autophagy and elevated expression of klotho protein, particularly in the kidney. Rapamycin treatment also increased cardiomyocyte Ca2+ -sensitivity and enhanced the rate constant of force re-development, which may also contribute to the enhanced survival rate in elderly mice.

Full open access paper:

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#2

Really interesting paper. Strongly implicates enhanced Klotho as a mechanism by which rapamycin affects mice. Pity they used so few mice and all males, but they measured several variables. I’m sure others will build on this work.
Makes me wonder if I’m doubling up as I take Sirolimus and inject Klotho…

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#3

Papers that make you go,“hmmm…”

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#4

Articles like this once again emphasize the fact that physical exercise is an essential component of any anti-aging strategy.

“High-intensity physical exercise increases serum a-klotho levels
in healthy volunteers”

https://journals.sagepub.com/doi/pdf/10.1177/1849454418794582

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#5

Be sure not to oversleep.

Conclusion

Our results revealed that people who sleep more than 7.5 hours per night have decreased levels of the anti-aging protein klotho in their serum, thus being more at risk of aging-related syndromes.

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#6

Absolutely!
I think I am keeping my Klotho levels up. My latest blood results seem to support this as measures of kidney health (creatinine, uric acid etc) are in the low range and in fact lower than previous tests. Klotho is a major factor in healthy kidney function. Sirolimus boosts Klotho.

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#7

The anti-aging protein Klotho has been associated with cardiovascular health protection. Nevertheless, the protective mechanism remains unknown. The present study is aimed at exploring the effect of Klotho on cardiac remodeling and its potential mechanism in mice with myocardial infarction (MI). We used left anterior coronary artery descending ligation to develop an MI model for in vivo analyses. In contrast, H9C2 cells and cardiac fibroblasts were used to establish the oxygen–glucose deprivation (OGD) model in in vitro analyses. In vivo and in vitro models were treated with Klotho. Compound C, an AMPK signaling inhibitor, was used to determine whether Klotho’s effects are mediated through the AMPK/mTOR signaling pathway. Echocardiography, Masson trichrome staining, immunofluorescence, immunohistochemistry, real-time polymerase chain reaction (RT-PCR), and western blot were used to detect the related indicators. The findings of the in vivo model indicate that Klotho treatment improved the mice’s cardiac function, reduced cardiac fibrosis, and attenuated myocardial inflammatory factors, ferroptosis, and oxidative stress.

Paywalled:

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#8

daily dose I suppose? most of the mice studies are based on daily dose and yet most human practice is weekly and bi-weekly.

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#9

Even if you gain the benefits weekly, I am sure it will make a major impact.

Just compare two individuals. One goes to the gym once a week, the other not at all. I can guess who would be healthier in the long run.

I also think taking Rapamycin has more of an impact than one gym workout.

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#10

No - daily dosing is for transplant patients. It reduces your immune system function. See this thread for dosing: What is the Rapamycin Dose / Dosage for Anti-Aging or Longevity?

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#11

Within four hours of getting the klotho injection, the monkeys given the lowest dose did significantly better on the memory tests than those given a placebo shot. The effect lasted for at least two weeks, after which the testing stopped.

In a surprising result, however, those given higher doses didn’t do better than the placebo group. That may be because the low dose was designed to be equivalent to the levels seen in humans at birth, says Dubal. “It might be that klotho needs to be just replenished to a certain level rather than be given at huge pharmacologic doses.”

“Given that most experiments in the ageing field employ short-lived animal models – like mice, flies and worms – it is impressive that the authors performed these experiments in a non-human primate,” says João Pedro de Magalhães at the University of Birmingham, UK. But researchers need to find out why klotho wasn’t beneficial at higher doses, he says.

Charles Brenner at City of Hope in Duarte, California, says the inconsistent results and the fact that the researchers weren’t always “blinded” as to which monkeys received klotho and which had the placebo suggest the results may be untrustworthy.

Longevity factor klotho enhances cognition in aged nonhuman primates

Cognitive dysfunction in aging is a major biomedical challenge. Whether treatment with klotho, a longevity factor, could enhance cognition in human-relevant models such as in nonhuman primates is unknown and represents a major knowledge gap in the path to therapeutics. We validated the rhesus form of the klotho protein in mice showing it increased synaptic plasticity and cognition. We then found that a single administration of low-dose, but not high-dose, klotho enhanced memory in aged nonhuman primates. Systemic low-dose klotho treatment may prove therapeutic in aging humans.

Open access paper link:

https://www.nature.com/articles/s43587-023-00441-x

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#12

For those people here who are interested in Klotho… a message from Steve Perry on a small trial he’s trying to do, perhaps people want to join him:

From: Steve “Endogenous” Perry <[steve@stevegperry.com]
Date: Tue, Jul 11, 2023 at 1:50 PM
Subject: One Injection of a Klotho Boosted Memory in Older Monkeys - it’s now time to do a GRG klotho study!

Hello,

Check out this Klotho article on Singularity Hub that originally appeared in the Journal of Nature. Best quote:

Four hours after administering the protein, the researchers subjected the macaques to a common memory test in which the animals have to remember where food has been hidden in an array of compartments. The monkeys were given two versions of the test, one of which was harder as it featured more compartments.

The team found that the monkeys’ performance on the easy test improved by about 6 percent compared to before the injection, but they did about 20 percent better on the hard one. The researchers continued to test the monkeys for the next two weeks and discovered that the cognitive boost persisted throughout this period.

Also the article goes on to say:

Proving that the therapy could work in humans will take a lot more research, but a Bay Area company called Unity Biotechnology has licensed the rights to UCSF’s work.

Well we are already on our way to proving klotho works in humans. As you may have heard, once GDF11ers down regulate, we offer them the option of taking .5 pg of Klotho per day. Right now, we have about 15 people taking klotho and almost all of them report a cognitive boost, usually in terms of writing and memory, about an hour after they take Klotho.

But you know me, I don’t like anecdotals like “cognitive boost”. My whole philosophy on curing aging is providing hard biomarker evidence that a regimen has efficacy. Therefore, I provide the charts below of my cognitive progress on GDF11 starting in 2014 and klotho starting in 2020. Note I have 3 years of cognitive baselines starting in 2010:

coghealth1
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reactiontime1
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The above charts are from my Physioage Analytics portal where over 800 of my biomarkers are tracked since 2007. The above charts are derived from my scores on the CNS vital signs test which I’ve been taking for over 13 years.

The first chart shows cognitive function score trending, which is a score weighted heavily on reaction time and processing speed as well as some verbal/visual memory scores. Since 2010, I have experienced a cognitive score increase from 3.25 to 3.67, a 13% increase. So no cognitive senescence for me.

The 2nd chart shows is just reaction time from 78 to 103, a 32% increase over 13 years. Note this is not the raw score in milliseconds, but a composite score CNS VS generates to be consistent with other scores and higher is better. Note that I started on GDF11 in 2014 and you can see a big jump from 74 to 95 from 2015 to a year later.

You may ask well what’s from GDF11 and what is from klotho? All GDF11ers see improvement in reaction time due to increased blood flow to the brain as well as increased neuronal autophagy. But klotho surely has helped get me to a record cognitive function score of 3.67. Though I will admit I have no data for klotho on its own.

To see how effective klotho is by itself, I propose we conduct a klotho study within the GRG. This should not be too hard since klotho, unlike GDF11, is pretty easy to dose. For anyone who is interested, I propose you do the following:

  1. Read my klotho paper on GDF11Rejuvenation.com under About.
  2. Buy two codes at a cost of $35 each, from CNSVS.com. Take a baseline CNS Vital Signs now and another one after 3 months on Klotho. Send me the PDF of your results and I will enter them into Physioage Analytics and send you a link so you can review your cognitive report card, trending of all CNS VS biomarkers, etc.
  3. You can purchase 25 ug for $218 klotho from BuckyLabs here. Also get two 30 ml vials of bacteriostatic water and dilute down to 1 unit = .1 pg. After baseline CNS VS, start dosing as per the klotho paper.
  4. Bonus points for those who put a CGM on their arm and track average glucose on klotho! Average glucose should come down pretty dramatically on klotho.

Are you in?

Steve

Steve Perry

CEO

GDF11Rejuvenation.com

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Injecting PF4 into the bloodstream
#13

Has anyone tried this?

#14

Neuroprotective Role of Klotho on Dementia

Klotho, a gene found on chromosome 13q12, is involved in a variety of processes and signaling pathways in the human body related to vitamin D metabolism; cardiovascular, renal, musculoskeletal, and skin diseases; and cancer biology. However, more importantly, it has been linked to beneficial effects related to anti-aging. The levels of soluble Klotho in the blood have been found to decline with age, increasing the risk of age-related diseases. When the Klotho gene was silenced or defective, it caused a shorter lifespan. However, when the gene was overexpressed, it resulted in a longer lifespan. Klotho has positive benefits on the neurological system by causing a higher representation of useful longevity genes, preventing further neuronal damage, and offering neuroprotection. Thus, it has the potential to become a new treatment for many age-related diseases that cause dementia, including multiple sclerosis, Alzheimer’s disease, and Parkinson’s disease. In this review, we discuss the mechanisms of Klotho’s benefits and roles on various organ systems, specifically on nervous system disorders that lead to dementia.

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