Older People Grow 2.5 Years Younger

Worldwide media coverage in September 2019 described publication of the first clinical trial to demonstrate significant reversals of biological aging in a study group of humans!

A major advance in healthy longevity potential was published on September 8, 2019, in the journal Aging Cell.¹

A combination of nutrients, hormones, and a drug resulted in significant human age reversal, as measured by DNA methylation, in nine men aged 51 to 65.

The study was conducted by Dr. Greg Fahy and colleagues at the biotechnology company Intervene Immune, Inc., in collaboration with researchers from Stanford University and the University of California, Los Angeles (UCLA).

The study protocol consisted of individualized doses of:

1. Human growth hormone (hGH),

2. DHEA, and

3. Metformin.

Study subjects were also provided with 3,000 IU of vitamin D3 and 50 mg of elemental zinc daily.

After one year, the study participants were 2.5 years younger, as measured by multiple DNA methylation aging tests, than they would have been without treatment.

This innovative protocol resulted in multiple, system-wide, anti-aging benefits:

1. Regenerated thymic structure,

2. Improved immune function with signs of increased cancer protection,

3. Improved prostate cancer markers (PSA and percent free PSA),

4. Regenerative effects on kidney function and bone marrow, and

5. On average, 2.5 years of age reversal, as measured by four different DNA methylation tests.

This study, called the TRIIM trial, captured worldwide media attention.

Favorable commentary was published in the prestigious journal Nature.²

It was the first topic of discussion on Joe Rogan’s podcast with Harvard scientist David Sinclair.³ (Joe Rogan reaches more people than many network TV shows.)

In fact, according to Altmetric,⁴ three months after publication, the paper was number four out of 1,455 in attention score (99^th percentile) among similar sources, and in the top 5% of all “research outputs” ever scored.

TRIIM stands for:

• Thymus Regeneration,
• Immunorestoration, and
• Insulin Mitigation = TRIIM

This combined protocol was designed to reach just the first two of the above goals.

The TRIIM trial results, therefore, were far greater than expected.

Regeneration of the Thymus

The thymus is a gland located behind the sternum (breastbone) and in front of your lungs, and it is responsible for the development of T-cells.

Beginning at the age of one, although the thymus continues to grow, it begins to become replaced with fatty tissue.⁶ The absolute mass of functional thymic tissue continues to increase until the early teenage years, but after that, progressive replacement by fat combined with a slow loss of thymic volume—a process known as thymic involution—results in a net decrease in thymic function. By the time we reach 50 years of age, the functional mass of the thymus is just a fraction of what it was at the onset of adolescence.⁷

The shrinkage, or involution, of the thymus has been correlated with:

1. Reductions in functional thymus-educated immune cells (T-cells),⁸

2. Reductions in T-cell receptors,⁹

3. Increased risk of several diseases,¹⁰ and

4. Increased risk of all-cause mortality.¹¹

Earlier research had shown that growth hormone treatment can regenerate the thymus gland in animals and relatively young immunodeficient HIV patients,^12-14 but the thymus of HIV patients is not normal. Until the TRIIM trial, hGH had never been used with the aim of reversing immunological aging in otherwise healthy, aging volunteers.

Growth Hormone Safety

Traditionally, there has been concern that growth hormone treatment can cause insulin resistance, leading to a damaging and pro-aging increase in insulin levels, and might stimulate proliferation of malignant cells,^15,16 although actual long-term population studies on hGH safety have not found an increased cancer risk.¹⁷

In the TRIIM trial, both problems were apparently successfully contained by the combination protocol employed in this study: insulin levels remained lower than average, and cancer risk seemed to decrease based on three independent indices of cancer risk.

Remarkably, all study subjects showed regeneration of their thymus, which was validated by MRI scanning using a special imaging technique at Stanford after 0, 9, 12, and sometimes also after five months of treatment. Regeneration of the thymus was reflected by the replacement of age-related non-functional fat tissue with functional, fat-free tissue.

Regeneration was so clear-cut that it was statistically significant at many time points in seven out of nine participants. Two others showed about a 10% increase in functional thymic mass, but the increase did not reach statistical significance because their baseline thymic fat contents were low for some reason. Overall, the probability that thymus regeneration did not occur was about one billionth of one billionth (p<9x10^-17), meaning that MRI evidence of thymic regeneration was incontrovertible.¹ (MRI scanning was funded in part by Life Extension^®.)

Immune System Benefits

Several blood tests revealed that there was significant improvement in many biomarkers of immune function from the treatment protocol. Notably, there was an increase in the production of new T-cells, indicating that the basic T-cell manufacturing function of the thymus had been reactivated.

Three classes of new or “naïve” T-cells were shown to increase. Since new T-cells normally seem to survive and function for many years, the hope is that these new immune defenders will continue to protect the TRIIM participants from immunological aging for many years to come.

The trial also uncovered the fact that, according to sophisticated test results, the treated study subjects had an increased lymphocyte to monocyte ratio. A higher lymphocyte to monocyte ratio (LMR) has been linked to better outcomes for many types of cancer, atherosclerosis, cardiovascular disease, stroke,^18-21 and all-cause mortality. Furthermore, the increase in LMR did not change even six months after treatment ended.¹

Dr. Fahy and his fellow researchers speculated that the benefits of having more lymphocytes in relation to monocytes might be due in large part to the fact that most monocytes express CD38, an enzyme that destroys NAD⁺ and may drive age-related NAD⁺ depletion in tissues.²² The LMR increase was driven mostly by a decrease in monocytes.

Although monocytes play an important role in “innate” immunity,²³ higher monocyte levels are associated with age-related frailty,²⁴ and CD38 expression increases as we age.

We need NAD⁺ for a myriad of biochemical processes including DNA repair and cell energy production.²⁵ Therefore, it may be that the TRIIM protocol achieves systemic age-reversing effects at least in part by limiting the levels of NAD⁺-depleting CD38 monocytes while also restoring youthful thymic immunity.

Reduced Cancer Risk Indicators

Immune function was also improved, as shown by a reduction in PD-1 expression on cytotoxic T-cells.¹

PD-1 is a receptor that cancer cells hijack to trick immune T-cells into thinking that the cancer is not a threat and they should not be attacked.²⁶

Drugs that block the PD-1 receptor such as Opdivo^® and Keytruda^® have become multi-billion-dollar blockbuster drugs in recent years due to the survival improvement they demonstrate against several different cancers.²⁷

Prostate cancer risk decreased in these study participants as shown through decreased PSA and improved percentage of free PSA.¹

Additionally, C-reactive protein (a marker for systemic inflammation) decreased significantly.¹

Elevated C-reactive protein is a biomarker for risk of both acute and chronic inflammatory conditions, including cardiovascular disease, surgical outcomes, and cancer mortality^28-32 and is an important marker for the generalized inflammation that normally accompanies aging (“inflammaging”).

Overall, these results support the hypothesis that improving thymic function can help to prevent cancer and augment defenses against the disease.

Kidney Function

One unexpected discovery was that there was a significant improvement in kidney function, as demonstrated by an increased glomerular filtration rate, or GFR.¹

Normally, kidney function never improves as aging proceeds, and end-stage renal failure now costs taxpayers more than $30 billion a year.

During the TRIIM treatment, GFR steadily improved, and the trend even seemed to continue for six months after discontinuing treatment. These results were one sign that perhaps the TRIIM treatment could influence systemic aging and might not be confined only to rejuvenation of the immune system.

Reversal of Epigenetic Age

Within the last six years or so, measurement of DNA methylation patterns has enabled the most accurate measurement of biological age presently available. These DNA methylation “clocks” are an exciting new development and are also allowing scientists to accurately predict future healthspan and lifespan.

Several different “clocks” of this kind have been developed, each with a somewhat different purpose in mind. In combination with other carefully selected clinical markers, they will become increasingly valuable predictors of future healthspan and mortality.^1,33,34

DNA Methylation Tests

DNA methylation influences how genes are expressed.

Healthy methylation allows healthy gene expression to support youthful cell function.

Unhealthy methylation patterns result in deleterious gene expression that facilitates the onset of degenerative illness and shortened lifespan.^35-37

As we age, our pattern of gene expression changes. By following changes in DNA methylation, we can indirectly follow changes in gene expression and, therefore, changes in biological age.

The first epigenetic aging “clock” was developed by UCLA researcher Dr. Steve Horvath, who assessed the methylation status of a broad array of tissue and cell types, finally settling on 353 specific DNA sites whose methylation is correlated with chronological age.

The TRIIM trial used Dr. Horvath’s original epigenetic clock, as well as three other methylation clocks, to allow a robust analysis of the results of the treatment, and to confirm that different epigenetic analyses agreed, on average, on the impact of the intervention.¹

Results of TRIIM Trial

By combining metformin and DHEA with growth hormone for just one year, the thymic benefits of growth hormone occurred with no significant side effects and the unexpected benefit of a 2.5-year biological age-reversal in all patients, according to an average of the four epigenetic clocks.¹

Remarkably, although these four clocks depend differently on blood composition and other factors, and although there were some quantitative differences in results between the different clocks, each clock produced results that were essentially the same as all the others with respect to epigenetic aging reversal.

Each clock, by itself, showed highly statistically significant results. In combination, the evidence for aging reversal across all clocks was overwhelming.

Interestingly, the average age reversal of 2.5 years was mirrored almost exactly by the original Horvath DNAm clock result. The DNAm clock for blood is known to correlate with aging of the brain, muscle, and other tissues, supporting a global aging reversal effect of the TRIIM protocol.

Summary

Regenerating thymic function, improving immunity, reversing epigenetic aging, and the other favorable effects seen in the TRIIM trial are required if we are to advance toward a more youthful state.

Dr. Greg Fahy’s company, Intervene Immune, Inc., is seeking to replicate and extend these results in a larger clinical trial that is expected to launch in the first quarter of 2020. The second phase of the clinical trial will not only have many more study subjects, but will also implement more tests, such as clinically significant and hopefully FDA-approvable health endpoints and will include women, minorities, older and younger participants, and those with imperfect health, as well as a control comparator group.

Life Extension^® has long advocated for use of several of the various interventions used in the TRIIM trial. These include DHEA and metformin. For those who don’t take metformin, we have discovered other AMPK-activating compounds. Most readers of this publication utilize these and other methods to support healthy, youthful immunity and DNA.

However, we have not previously recommended hGH, or the combination of hGH, metformin, and DHEA, which breaks new ground, and we welcome this advance.

Dr. Fahy’s group is seeking to identify an affordable source of growth hormone as well as other ways to make this longevity protocol available to almost everyone. For more information, see www.interveneimmune.com.

We applaud Dr. Greg Fahy, his team, and all the researchers involved for their arduous and excellent work in this historical clinical trial!

Look forward to future updates!

If you have any questions on the scientific content of this article, please call a Life Extension^® Wellness Specialist at 1-866-864-3027.

References

1. Fahy GM, Brooke RT, Watson JP, et al. Reversal of epigenetic aging and immunosenescent trends in humans. Aging Cell. 2019;18(6):e13028.
2. Abbott A. First hint that body’s ‘biological age’ can be reversed. Nature. 2019 Sep;573(7773):173.
3. Available at: https://www.youtube.com/watch?v=ZGLL77wYxe8. Accessed November 29, 2019.
4. Available at: https://wiley.altmetric.com/details/66240556. Accessed December 12, 2019.
5. Available at: https://www.forbes.com/sites/robinseatonjefferson/2019/09/09/cocktail-of-drugs-gives-first-hope-that-biological-age-can-be-reversed/. Accessed November 29, 2019.
6. Palmer S, Albergante L, Blackburn CC, et al. Thymic involution and rising disease incidence with age. Proceedings of the National Academy of Sciences. 2018;115(8):1883-8.
7. Rezzani R, Nardo L, Favero G, et al. Thymus and aging: morphological, radiological, and functional overview. Age (Dordr). 2014 Feb;36(1):313-51.
8. Arnold CR, Wolf J, Brunner S, et al. Gain and loss of T cell subsets in old age—age-related reshaping of the T cell repertoire. J Clin Immunol. 2011 Apr;31(2):137-46.
9. Naylor K, Li G, Vallejo AN, et al. The influence of age on T cell generation and TCR diversity. J Immunol. 2005 Jun 1;174(11):7446-52.
10. Ventevogel MS, Sempowski GD. Thymic rejuvenation and aging. Curr Opin Immunol. 2013 Aug;25(4):516-22.
11. Ferrando-Martinez S, Romero-Sanchez MC, Solana R, et al. Thymic function failure and C-reactive protein levels are independent predictors of all-cause mortality in healthy elderly humans. Age (Dordr). 2013 Feb;35(1):251-9.
12. Napolitano LA, Lo JC, Gotway MB, et al. Increased thymic mass and circulating naive CD4 T cells in HIV-1-infected adults treated with growth hormone. Aids. 2002 May 24;16(8):1103-11.
13. Taub DD, Murphy WJ, Longo DL. Rejuvenation of the aging thymus: growth hormone-mediated and ghrelin-mediated signaling pathways. Current opinion in pharmacology. 2010;10(4):408-24.
14. Savino W. Neuroendocrine control of T cell development in mammals: role of growth hormone in modulating thymocyte migration. Exp Physiol. 2007 Sep;92(5):813-7.
15. Harman SM, Blackman MR. Use of growth hormone for prevention or treatment of effects of aging. J Gerontol A Biol Sci Med Sci. 2004 Jul;59(7):652-8.
16. Marcus R, Butterfield G, Holloway L, et al. Effects of short term administration of recombinant human growth hormone to elderly people. J Clin Endocrinol Metab. 1990 Feb;70(2):519-27.
17. Svensson J, Bengtsson BA. Safety aspects of GH replacement. Eur J Endocrinol. 2009 Nov;161 Suppl 1:S65-74.
18. Gong S, Gao X, Xu F, et al. Association of lymphocyte to monocyte ratio with severity of coronary artery disease. Medicine. 2018;97(43):e12813-e.
19. Wang Q, Ma J, Jiang Z, et al. Association of lymphocyte-to-monocyte ratio with in-hospital and long-term major adverse cardiac and cerebrovascular events in patients with ST-elevated myocardial infarction. Medicine. 2017;96(34):e7897-e.
20. Hu P, Shen H, Wang G, et al. Prognostic significance of systemic inflammation-based lymphocyte- monocyte ratio in patients with lung cancer: based on a large cohort study. PLoS One. 2014;9(9):e108062.
21. Caglayan V, Onen E, Avci S, et al. Lymphocyte-to-monocyte ratio is a valuable marker to predict prostate cancer in patients with prostate specific antigen between 4 and 10 ng/dl. Arch Ital Urol Androl. 2019 Jan 17;90(4):270-5.
22. Camacho-Pereira J, Tarrago MG, Chini CCS, et al. CD38 Dictates Age-Related NAD Decline and Mitochondrial Dysfunction through an SIRT3-Dependent Mechanism. Cell Metab. 2016 Jun 14;23(6):1127-39.
23. Yang J, Zhang L, Yu C, et al. Monocyte and macrophage differentiation: circulation inflammatory monocyte as biomarker for inflammatory diseases. Biomarker Research. 2014 2014/01/07;2(1):1.
24. Samson LD, Boots AMH, Verschuren WMM, et al. Frailty is associated with elevated CRP trajectories and higher numbers of neutrophils and monocytes. Exp Gerontol. 2019 Oct 1;125:110674.
25. Johnson S, Imai SI. NAD (+) biosynthesis, aging, and disease. F1000Res. 2018;7:132.
26. Salmaninejad A, Valilou SF, Shabgah AG, et al. PD-1/PD-L1 pathway: Basic biology and role in cancer immunotherapy. J Cell Physiol. 2019 Aug;234(10):16824-37.
27. Ardolino L, Joshua A. Immune checkpoint inhibitors in malignancy. Australian prescriber. 2019;42(2):62-7.
28. Suzuki S, Akiyoshi T, Oba K, et al. Comprehensive Comparative Analysis of Prognostic Value of Systemic Inflammatory Biomarkers for Patients with Stage II/III Colon Cancer. Ann Surg Oncol. 2019 Nov 12.
29. Bouassida M, Zribi S, Krimi B, et al. C-reactive Protein Is the Best Biomarker to Predict Advanced Acute Cholecystitis and Conversion to Open Surgery. A Prospective Cohort Study of 556 Cases. J Gastrointest Surg. 2019 Nov 25.
30. Straatman J, Harmsen AM, Cuesta MA, et al. Predictive Value of C-Reactive Protein for Major Complications after Major Abdominal Surgery: A Systematic Review and Pooled-Analysis. PLoS One. 2015;10(7):e0132995.
31. Available at: https://www.hopkinsmedicine.org/heart_vascular_institute/clinical_services/centers_excellence/womens_cardiovascular_health_center/patient_information/health_topics/c_reactive_protein.html. Accessed December 4, 2019.
32. Ridker PM. A Test in Context: High-Sensitivity C-Reactive Protein. J Am Coll Cardiol. 2016 Feb 16;67(6):712-23.
33. Levine ME, Lu AT, Quach A, et al. An epigenetic biomarker of aging for lifespan and healthspan. Aging (Albany NY). 2018 Apr 18;10(4):573-91.
34. Lu AT, Quach A, Wilson JG, et al. DNA methylation GrimAge strongly predicts lifespan and healthspan. Aging. 2019;11(2):303-27.
35. Johnson AA, Akman K, Calimport SRG, et al. The role of DNA methylation in aging, rejuvenation, and age-related disease. Rejuvenation research. 2012;15(5):483-94.
36. Xiao FH, Kong QP, Perry B, et al. Progress on the role of DNA methylation in aging and longevity. Brief Funct Genomics. 2016 Nov;15(6):454-9.
37. Mahmoud AM, Ali MM. Methyl Donor Micronutrients that Modify DNA Methylation and Cancer Outcome. Nutrients. 2019;11(3):608.