As the first post on this forum, I'd like to state that we are all prisoners of our own bias. We are shaped by the experiences we have had and the knowledge we have accumulated in our careers. As a self proclaimed "expert", I readily admit I don't have the pedigree of much more accomplished researchers, but I do have 25 years of entrepreneurial experience evaluating new technologies around stem cells and their applications. Many ideas in this area build upon others and we all stand on the shoulders of our predecessor giants. So I will present my analysis of this paper, and then I will ask the same question to the ChatGTP AI, and present them side by side.
I first learned of this paper through Tik Tok, as its algorithm is excellent in screening for topics I’m interested in, and it naturally got my attention to investigate further.
The paper referenced is here https://www.aging-us.com/article/204896/pdf
ABSTRACT
A hallmark of eukaryotic aging is a loss of epigenetic information, a process that can be reversed. We have previously shown that the ectopic induction of the Yamanaka factors OCT4, SOX2, and KLF4 (OSK) in mammals can restore youthful DNA methylation patterns, transcript profiles, and tissue function, without erasing cellular identity, a process that requires active DNA demethylation. To screen for molecules that reverse cellular aging and rejuvenate human cells without altering the genome, we developed high-throughput cell-based assays that distinguish young from old and senescent cells, including transcription-based aging clocks and a real-time nucleocytoplasmic compartmentalization (NCC) assay. We identify six chemical cocktails, which, in less than a week and without compromising cellular identity, restore a youthful genome-wide transcript profile and reverse transcriptomic age. Thus, rejuvenation by age reversal can be achieved, not only by genetic, but also chemical means.
My first thoughts upon reading the abstract is that OCT4 and SOX2 are well studied, roughly 10,000 papers each in PubMed. So what is in this paper that’s so unique? Second thought, is DNA methylation the right metric to measure cellular aging, and is it clinically relevant here?
So let’s dive in. One of the best analogies I’ve heard for distinguishing between genetic and epigenetic information is that if genetic information are notes on the musical staff, then epigenetic information is the tablature above the notes. It’s what your body does with that genetic information that makes it specific to you, and we all have our own music playing.
For all you conspiracy theorists out there, the authors acknowledge the inherent dangers of delivering RNA cellular reprogramming via lipid nano particles, and the strategy they present here is to produce a chemical cocktail to reprogram cells to a pluripotent state without the need for DNA/RNA delivery. (No need for mandated and untested gene therapies.)
THE ASSAY Nucleocytoplasmic compartmentalization (NCC) Actually, it’s the first time I’ve seen this assay presented as part of anti aging studies, but seems legit, and is similar to other senescence based fluorescent assays. So I had to jump ahead a little bit, and figure out where they were going, as a lot of the paper is in defense of THE ASSAY.
I don’t have any reason to disbelieve it’s an appropriate assay, it is Harvard and MIT for God’s sake.
So they use the NCC assay to find out what’s upregulated/downregulateed and unchanged. The baselines are a 14 year old and a 94 year old to basically calibrate the assay to what you are trying to test, so the following is what they tested.
SO HERE IS WHATS IN THE COCKTAIL
V – VALPROIC ACID
C – CHIR-99021 (A GSK3 inhibitor that promotes cell growth)
N – TTNPB(N) (Retinoid pathway activator)
Y – Y-2763 (Inhibitor of Rho-associated, coiled-coil containing protein kinase (ROCK))
S – SMOOTHENED AGONIST
A – ABT-869(A) (Tyrosine Kinase inhibitor)
6 - E-616452 (transforming growth factor-beta receptor, kinase 5 (TGF-β-RI/ALK5) inhibitor.)
T- TRANYLCYPROMINE
F- FORSKOLIN
OTHER SPICES; SODIUM BUTYRATE, (bFGF), ALPHA-KETOGLUTURATE
So they knew the combinations of VCT6T andVCT6F could make old cells into stem cells without DNA or a plasmid being transfected into the cell. In other words, the right chemical cocktail induced the cells to change into pluripotent or what most people call stem cells. Of the 80 cocktails tested, the VC6TF cocktail was the most effective in the assay. 6 cocktails were identified and used the spices to enhance efficacy. I had to read the paper again in its entirety to understand they never divulge what’s in the 6 cocktails they tested, only the results in THE ASSAY.
The discussion section accurately describes the limitations of the technology, additional testing in mammalian models, etc. The hope would be to take your cells; saliva, blood, epithelial, etc, and subject them to the cocktail while they are amplified in a lab. The cells could be returned to you without fear of rejection or tumorgenesis. I don’t think it’s ever a pill as it would have to pass the gastrointestinal tract, and from what I know of cellular replication, it takes the entire cocktail for the cells to be suspended in as they replicate.
ChatGPT response to epigenetic reprogramming and the Harvard cocktail
Of course, I'd be happy to elaborate further on the topic of epigenetic reprogramming and the "Harvard cocktail." The term "Harvard cocktail" is often used to refer to a combination of small molecules that was developed by researchers at Harvard University to induce epigenetic reprogramming in cells. This cocktail is intended to reprogram somatic (differentiated) cells back into induced pluripotent stem cells (iPSCs) by erasing their existing epigenetic marks and returning them to a more pluripotent state. iPSCs are cells that closely resemble embryonic stem cells in their ability to differentiate into various cell types, making them a valuable resource for regenerative medicine and disease modeling. The cocktail typically consists of various small molecules that target key epigenetic regulators, such as DNA methyltransferase inhibitors, histone deacetylase inhibitors, and other compounds that influence chromatin remodeling. These molecules collectively work to modify the epigenetic landscape of the cells, potentially allowing them to regain pluripotent characteristics. The goal is to reverse the differentiation process that cells have undergone and guide them to a state where they can differentiate into a wide range of cell types. However, while the idea of using the Harvard cocktail and similar approaches to induce epigenetic reprogramming is intriguing, there are several challenges and considerations to keep in mind:
1. Efficiency and Specificity: Epigenetic reprogramming is a complex process, and achieving consistent and efficient conversion of somatic cells to pluripotent states using small molecules is still a challenge. Additionally, achieving precise control over the extent of reprogramming and ensuring that the desired cell types are generated is crucial. 2. Safety and Quality Control: The use of small molecules for epigenetic reprogramming could lead to unintended effects on gene expression, potentially resulting in aberrant cell behavior or tumorigenicity. Ensuring the safety and stability of the reprogrammed cells is a critical aspect of this research. 3. Cellular Heterogeneity: Differentiated cells within tissues are often heterogeneous, and not all cells may respond uniformly to the reprogramming cocktail. Some cells might not fully reset their epigenetic marks or might require additional manipulation to achieve the desired pluripotent state. 4. Alternative Approaches: The Harvard cocktail approach is just one of several strategies for epigenetic reprogramming. Other methods, such as genetic manipulation or direct protein delivery, have also been explored to achieve similar outcomes. 5. Ethical Considerations: The potential to generate pluripotent stem cells through epigenetic reprogramming raises ethical considerations related to their use in research and potential clinical applications.
In summary, while the concept of using a "Harvard cocktail" or similar small molecule combinations for epigenetic reprogramming is promising, it's important to recognize that this field is still evolving. Researchers continue to refine the protocols, understand the underlying mechanisms, and address the challenges associated with inducing epigenetic changes in cells. As our knowledge advances, we may see more precise and effective methods for cellular reprogramming emerge, holding significant potential for regenerative medicine and disease treatment.