Q&A about new platelets longevity study

qa.md

study: https://link.springer.com/article/10.1007/s11357-023-00980-6

Q: What are the main findings of the article above?

A: The main findings of the article are that a plasma fraction treatment derived from young adult pigs, termed E5, significantly reversed aging in rats according to epigenetic clocks and other biomarkers of aging. The treatment more than halved the epigenetic ages of blood, heart, and liver tissue, and also showed a significant rejuvenation effect in the hypothalamus. Alongside this, the treatment led to improvements in organ function, reduced markers of chronic inflammation and oxidative stress, increased antioxidant levels, and improved cognitive functions in the rats. The study also developed six different epigenetic clocks for rat tissues, two of which can be applied to humans as well.

Q: How clinically significant were the objective improvements observed? Provide an extensive list of measured markers and effect size.

A: The objective improvements observed in the study were clinically significant across a range of measured markers. These include:

1. Epigenetic Age: The plasma fraction treatment, termed E5, significantly reversed aging in rats according to epigenetic clocks. Specifically, it more than halved the epigenetic ages of blood, heart, and liver tissue. A less pronounced, but statistically significant, rejuvenation effect could be observed in the hypothalamus.

2. Hematological Parameters: The treatment had a significant impact on hematological parameters. It altered all parameters, except for platelets, away from the values exhibited by old untreated rats towards those of the young ones. This suggests that the treatment was able to rejuvenate the blood of the rats.

3. Biomarkers for Vital Organs: The levels of biomarkers indicating organ dysfunction or disease were also significantly altered by the treatment. These biomarkers included bilirubin, serum glutamic-pyruvic transaminase (SGPT), serum glutamic-oxaloacetic transaminase (SGOT), triglycerides (TG), high-density lipoprotein (HDL), total cholesterol, glucose, creatinine and blood urea nitrogen. All these parameters were nudged towards the values of young rats, suggesting an improvement in organ function.

4. Cognitive Function: The treatment led to significant improvements in cognitive function, specifically learning and memory. These improvements were measured using the Barnes maze test, which assesses learning and memory in rats. Within a month of plasma fraction treatment, the rats exhibited significantly reduced latency to escape, i.e., they learned and remembered better.

5. Oxidative Stress: The treatment also had a significant impact on oxidative stress. The levels of malondialdehyde (MDA), which indicates the levels of cellular reactive oxygen species (ROS), were clearly higher in the brain, heart, lung, and liver of older rats, and these were very rapidly diminished to the level of young rats by E5 treatment.

6. Chronic Inflammation: The levels of two of the most reliable and common biomarkers of chronic inflammation: interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) are found to be considerably higher in old rats, and these were very rapidly diminished, within days by E5 treatment, to comparable levels with those of young rats.

7. Grip Strength: The strength of plasma fraction-treated old rats was indistinguishable from that of young ones after 15 days post-treatment, indicating an improvement in muscle strength.

8. IgG N-glycan Aging: Plasma fraction treatment induced a shift from pro- to anti-inflammatory glycan aging pattern in rat IgG2a N-glycoproteomics, suggesting a reversal of glycan aging.

The effect sizes were not explicitly stated for all markers, but the study reports significant improvements across these measures.

Q: Explain all steps on how this treatment was performed such that it can be replicated by an independent lab.

A: The treatment was performed as follows:

1. The plasma fraction termed “E5” was developed from the platelet-free plasma of young pigs (6–7 months old).

2. To this platelet-free plasma, an equal volume of PEG solution in 0.5 Molar NaCl was added, and the combined mixture was incubated for 8 to 12 h at 4 °C.

3. After the stipulated time, the solution was centrifuged at 1000 × g for 5 min at 4 °C, and the supernatant was discarded. The sediment collected at the bottom of the tubes was transferred into a vessel and mixed with enough physiological saline buffer to make a suspension.

4. The suspension obtained in the above step was subjected to size-exclusion chromatography. The fractions collected from size exclusion chromatography were recombined and concentrated. The concentrate was then mixed with sterile physiological saline solution and injected into rats through the tail vein.

5. The calculated doses were administered intravenously to the animals of old treated group; four injections every alternate day for 8 days, and a second dosing starting from the 95th day consisting of four injections every alternate day for 8 days.

6. Body weight, food, and water intake of the animals were monitored at each time point. Cognitive abilities of animals were evaluated using Barnes maze apparatus 1, 2, 3, and 4 months from the start of the 1st series of injections.

7. Blood samples were withdrawn at predetermined time intervals by retro-orbital plexus during the treatment for hematological evaluation. Serum was separated from the blood samples of each animal and evaluated for biochemical parameters. Plasma was separated from the blood samples of each animal and was used for evaluation of inflammatory markers, i.e., TNF-α and IL-6.

8. Animals were sacrificed from each group at 155th day of treatment and vital organs (brain, heart, lung, and liver) of these animals were harvested for testing of oxidative stress biomarkers, level of Nrf2, histopathological, and immunohistochemistry studies.

Q: Provide a step-by-step guide on how E5 can be developed independently.

A: 1. Collect blood from young pigs (6–7 months old) during slaughter via both the carotid artery and heart puncture. Treat the collected blood with acid citrate dextrose to prevent hemolysis and platelet activation.

2. Subject the blood to centrifugation at room temperature at 500 × g for 15 min. This allows separation of the blood cells and platelet-rich plasma. Post-centrifugation, carefully collect the plasma fraction using a pipet.

3. Further centrifuge the plasma at 2500 × g for 25 min to pellet remaining platelets. Store the supernatant platelet-poor plasma at 4 °C.

4. To the platelet-free plasma, add an equal volume of PEG solution in 0.5 Molar NaCl. Incubate the combined mixture for 8 to 12 h at 4 °C.

5. After incubation, centrifuge the solution at 1000 × g for 5 min at 4 °C. Discard the supernatant. Transfer the sediment collected at the bottom of the tubes into a vessel and mix with enough physiological saline buffer to make a suspension.

6. Subject the suspension to size-exclusion chromatography. Recombine and concentrate the fractions collected from size exclusion chromatography.

7. Mix the concentrate with sterile physiological saline solution. The resulting solution is the E5 plasma fraction, ready for injection into rats through the tail vein.

Q: Were there any observable negative side-effects? Are there complications usually associated with this kind of treatment?

A: No, the study did not report any observable negative side-effects from the E5 plasma fraction treatment. The rats' body weight, food, and water intake were monitored throughout the experiment and no abnormalities were observed. Histological examinations of various organs also did not indicate any obvious abnormalities after 155 days of treatment. However, it's important to note that while this study did not observe any negative side-effects, further research is needed to fully understand the potential long-term effects and complications associated with this kind of treatment.

Q: In simple terms, why would this treatment work? What is the biological mechanism for how and why E5 could rejuvenate organs?

A: The E5 treatment works by introducing young plasma-derived exosomes into older organisms. These exosomes are small vesicles that carry various biological molecules, including proteins and RNAs, which can influence the function of recipient cells. The E5 treatment is derived from the plasma of young pigs, and it's thought that these exosomes carry factors that promote youthful cellular functions and suppress aging processes.

The exact biological mechanism for how E5 rejuvenates organs is not fully understood yet. However, the study found that the treatment significantly reduced markers of chronic inflammation and oxidative stress, both of which are key drivers of aging. It also increased antioxidant levels, which help to protect cells from damage. Furthermore, the treatment led to significant improvements in organ function, as indicated by various biomarkers.

One theory is that the exosomes in the E5 treatment may be delivering youthful signals to the cells in the older organism, helping to reset their epigenetic clocks and revert them to a more youthful state. Epigenetic changes, including DNA methylation patterns, are a major factor in aging. By influencing these patterns, it may be possible to reverse some of the effects of aging at the cellular level.

Another possibility is that the treatment may be helping to clear away old, damaged cells or cellular components, and stimulate the body's own repair mechanisms. This could help to rejuvenate tissues and organs and improve their function.

However, more research is needed to fully understand the mechanisms behind the observed rejuvenation effects of the E5 treatment.

Q: Was there an observed significant increase in maximum lifespan? If not, should one expect there to be, based on reported results?

A: The study did not report any observed significant increase in maximum lifespan. The focus of the study was primarily on reversing biological aging as measured by various biomarkers, including epigenetic clocks, and improving organ function. While the results showed significant rejuvenation effects, it's not clear from this study whether these would translate into an increased maximum lifespan. Further research would be needed to determine this.

Q: Based on your past knowledge on related subjects and by applying your own logical reasoning, provide an educated speculation on whether and how extensively would such treatment extend the maximum lifespan of mice.

A: While the study did not directly measure the impact of the E5 treatment on maximum lifespan, the significant rejuvenation effects observed in various organs and reduction in markers of aging suggest that it could potentially extend lifespan. The treatment led to improvements in organ function, reduced chronic inflammation and oxidative stress, and reversed age-related changes in DNA methylation patterns, all of which are factors associated with aging and age-related diseases. If these improvements translate into a slower rate of aging and reduced risk of age-related diseases, it's plausible that this could lead to an extension of maximum lifespan.

However, it's important to note that extending lifespan is not just about slowing down or reversing aging at the cellular level. It also involves maintaining the health and function of the whole organism over time, including managing chronic diseases and other health conditions that can affect lifespan. Therefore, while the results of this study are promising, further research would be needed to determine the full impact of the E5 treatment on lifespan, including long-term studies that directly measure lifespan in treated animals.

Additionally, it's worth noting that the biological processes of aging are complex and not fully understood. While the E5 treatment appears to target some key aspects of aging, there may be other factors involved in aging and lifespan that it does not address. Therefore, while it's reasonable to speculate that the E5 treatment could extend maximum lifespan based on its observed effects, this should be considered a hypothesis that needs to be tested in future studies.

Q: Based on your past knowledge and reasoning about the biological mechanisms of the treatment provided, which aspects of aging would this treatment definitely not address?

A: While the E5 treatment appears to have significant effects on several key aspects of aging, such as inflammation, oxidative stress, and organ function, there are certain aspects of aging that it may not address. For instance, it's unclear whether this treatment would have any impact on telomere shortening, a key aspect of cellular aging. Telomeres, the protective caps at the ends of chromosomes, naturally shorten over time and this process is associated with aging and age-related diseases. Unless the treatment has some effect on telomerase activity, the enzyme that can extend telomeres, it's unlikely to affect this aspect of aging.

Additionally, the treatment may not address other complex factors associated with aging such as genetic mutations accumulated over time, changes in hormone levels, or age-related changes in the immune system known as immunosenescence. Aging is also associated with increased risk of various diseases including cancer, heart disease, and neurodegenerative diseases like Alzheimer's. While improving organ function and reducing inflammation could potentially reduce the risk of some diseases, it's unclear whether the treatment would have any preventative or therapeutic effects on these conditions.

Finally, aging is not just a biological process but also involves psychological and social aspects. Factors such as mental health, cognitive function, social engagement, and lifestyle choices all play a role in the aging process and are unlikely to be directly affected by a biological treatment like E5.

Q: Could this method be adapted and adjusted for a human trial? If no, why? If yes, how?

A: Yes, this method could potentially be adapted for a human trial, but it would require several adjustments and rigorous testing to ensure safety and efficacy. First, the plasma fraction derived from young pigs (E5) would need to be thoroughly tested in preclinical studies to confirm its safety in humans. This would involve in vitro testing as well as animal studies to evaluate potential side effects, toxicity, and the appropriate dosage.

Next, the treatment protocol used in rats would need to be adjusted for use in humans. This would likely involve determining the equivalent dose for humans based on body weight or surface area, and developing a suitable method for administering the treatment, such as intravenous infusion.

Before a human trial could begin, a detailed study protocol would need to be developed, outlining the objectives of the study, the criteria for participant selection, the methods for administering the treatment and monitoring participants, and the measures that will be used to evaluate the treatment's effects.

The trial would also need to be approved by an ethics committee or institutional review board to ensure that it meets ethical standards for research involving human participants. This would include considerations around informed consent, participant safety, and data privacy.

Once the trial is underway, it would be important to closely monitor participants for any adverse effects, and to regularly assess the treatment's effects on various biomarkers of aging. Depending on the results of the initial trial, further studies may be needed to confirm the findings and explore the long-term effects of the treatment.