Do Current Stem Cell Therapies Produce Rejuvenation?

Here is an interesting question for today's discussion: do present stem cell therapies produce results that we might in any way classify as rejuvenation? The therapies presently available in numerous clinics around the world vary in the type and quality of cells used, and whether they are derived from the patient's tissues. Simple approaches extract cells from fat tissue or cord blood or similar sources and use limited purification to enrich the proportion of stem cells and progenitor cells of various types. More sophisticated approaches standardize to proven, narrow methodologies for a single cell type at high purity levels, such as the widely used mesenchymal stem cells. Further variants involve the addition of scaffolds, nutrient gels, or adjuvant treatments to try to keep the transplanted cells alive and doing their thing for longer. The evidence gathered to date strongly suggests that near all of the stem cell transplant therapies deployed over the past fifteen years produce benefits through signaling: the transplanted cells don't as a rule hang around for long, but they alter the behavior of native cells, such as stem cells and immune cells.

This is a far cry from the class of stem cell treatment envisaged for the future of rejuvenation treatments after the SENS model. That would include the delivery of populations of engineered stem cells, derived from the patient, but with their age-related molecular damage removed. The intention would be for these cells to take up residence, possibly at the same time as the existing damaged stem cell populations are cleared out. It may involve repair of the stem cell niche as well, a more complex and daunting prospect given the complexities of the niches that are well understood. Many more are not, and even if simply delivering new stem cells for the long term, something that is presently beyond the state of the art, there are scores of such cell types. Each will require their own special handling, if the challenges seen so far in regenerative medicine continue to hold true. The reason that only a few cell types are widely used in today's stem cell treatments is that each cell type requires a very different recipe and methodology. It has taken a great deal of time and effort to arrive at the recipes presently in circulation. This SENS approach to stem cell replacement is clearly rejuvenation, however. The old is cast out and the youthful ushered back in.

The closest thing to a SENS-style stem cell therapy that has taken place are the immune system reboot treatments for type 1 diabetes and multiple sclerosis. Chemotherapy is used to kill off the old immune cells and stem cell therapy delivers a new set of cells, albeit not a set of cells with any age-related damage removed. Chemotherapy is an unpleasant thing to go through, but it actually worked. This is quite an old approach in comparison to much of what is discussed here; if you go digging you'll see that it was even attempted with mixed results for rheumatoid arthritis long enough ago to predate the advent of the modern standard treatment of biologics for immune suppression. The need for aggressive chemotherapy seems to have discouraged adoption of this approach, at least until targeted cell killing is a safer, less unpleasant undertaking.

Is this rejuvenation, however? Neither type 1 diabetes nor multiple sclerosis are age-related diseases; they are not a part of aging, though like all non-age-related disease they interact with aspects of aging in ways that are never good for the patient. I think we can all agree that fixing a broken leg isn't rejuvenation, and for the same reasons neither is fixing an immune system that breaks because of inherited genetic mutation, inflammatory injury, or simple bad luck. How about the rest of the modern panoply of stem cell therapies, the much more common and straightforward transplantation to enhance regeneration? These treatments produce benefits by providing a temporary period of (a) increased healing, sometimes regeneration that would never normally happen, such as in heart tissue, in other cases a matter of restoring more youthful levels of healing, (b) reductions in chronic inflammation via interaction with immune cells, and (c) other, less well cataloged changes such as lowered oxidative stress in tissues.

Perhaps the most common form of present day stem cell therapy are those intended to partially address joint wear in the old, such as early osteoarthritis, and the less dramatic but still potentially debilitating cases of middle-aged muscle and tendon damage. There is a high expectation of attaining modest benefits in terms of reduced pain and increased capacity for use, the risks are minimal, and the costs are reasonable - as little as a few thousand dollars with some footwork, even in the broken US medical system. You probably know someone who has investigated the treatment, and if not today then you will a few years from now. If an age-worn joint is marginally improved, but via the actions of your old cells, shoring up old tissues with more age-damaged cells, is that rejuvenation? You are better off, but the joint is no younger in any biochemical measure of aging. I can see the way to argue either side of that question with little difficulty, but on the whole I'm inclined to say that if there is room for debate then the results are not rejuvenation - and thus we're not close enough to the point in the road ahead where we can relax.

Comments

and thus we're not close enough to the point in the road ahead where we can relax.

Who is relaxing?

Posted by: Daniel Lemire at January 20th, 2016 9:39 PM

@Reason

Hi Reason !
Not to reduce the positive and hopeful message of stem cell injection.
Most everyone is dubious on stem cell rejuvenation, it will improve health and average lifespan. Mesenchymal stem cells and adipose stem cells increase lifespan in mice by rebuilding tissues and in certain studies they use stem cells that have no damage at all, they are extracted from embryos or foetus, at the very youngest point; yet, when they are transplanted in old mice they rejuvenate tissues and heal tissue damage partially. They do not increase MLSP all that much or not all. Centenarians maintain their stem cell niche longer, but this is due to lifelong maintenance of
*non-rising* stable low levels of oxidative stress markers, thus lifelong low oxidative damage to their stem cells..
This means they act mostly on pathological aging, not much on replicative redox-based and telomeres-based autophagic intrinsic aging.
It is a health rejuvenation not a true biorejuvenation to youth phenotype state reversal or true aging in reversal: Deaging (back to young self, like Benjamin Button effect). Stem cells sadly don't stop intrinsic-dependent damages all that much to matter over a very long lifespan of damage accumulation.

Here is true lifespan rejuvenation in a study in naked mole rat (lives 10 Times longer than a mouse, 30 vs 3 years, that qualifies as Extreme MLSP Lifespan Extension "Rejuvenation") that demonstrates the power of the redox to alter autophagic/proteasomal clearance of unfolded proteins, reduce age pigments in post-mitotic long-lived cells of long-lived mammals through reduced oxidative stress telomeres DNA damage, delaying replicative senescence onset and thus, intrinsic aging, by lifelong maintenance (no age-related loss) of redox homeostasis.

''Compared with data from young mice, young MRs [Naked Mole Rats] have 1.6 times as much free protein thiols groups and similar amounts of reversible oxidative damage to cysteine.
**In addition, they show less urea-induced protein unfolding, less protein ubiquitination and higher
proteasome activity. Mice show a significant age-related increase in cysteine oxidation and higher levels of ubiquitination.**
****In contrast, none of these parameters were significantly altered - over 2 decades****.
2 Decades that's 20 Years, 2-0-y-e-a-r-sssss that's Longgg by research model standards..that beats any kind of stem cell injection in mice and Any existing therapy or not. Stem cell injection could give a proportinate average lifespan extension -in humans in a ballpark figure of 10 or 15 years by increasing quality (maintenance of) healthspan (pushing humans closer and closer to their specie maximum lifespan potential), nice but nothing of true biorejuvenation intrinsic phenotype deaging; thus, humans will never surpass MLSP on stem cells because of no effect on intrinsic aging process.

Protein stability and resistance to oxidative stress are determinants of longevity in the longest-living rodent, the naked mole rat
1. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2651236

Posted by: CANanonymity at January 20th, 2016 10:38 PM

@Daniel Lemire: There has long been a sizable contingent of folk in the longevity advocacy community who think that stem cell medicine has the potential to fix most problems in aging. Sadly not the case, of course.

Posted by: Reason at January 21st, 2016 8:06 AM

I’d like to see the following experiment done using an aged mouse:

The basic concept is to locally or systemically restore “youthful” signaling in aged tissues (inspired by Conway’s experiments) using implants containing genetically engineered cells.

Extract fibroblasts and form an iPSC culture, reverting the cells to a fetal stage. The iPSC culture will provide the starting cells for genetic engineering.

Genetically engineer the cells to produce transcription factors maintaining the cells in the desired differentiation state while releasing molecules that rejuvenate aged body tissues. In later experiments further gene engineering could remove DNA defects and/or produce additional “youth” signaling molecules. Different tissues might require different genetic alterations.

Encapsulate the engineered cells in a removable implant that allows passage of nutrients and signaling molecules but otherwise prevents the engineered cells from unwanted interaction with the body. Hopefully the engineered cells in the implant would produce signaling molecules and targeted exosomes which rejuvenate body tissues, including stem cells.

The implants could be placed in specific locations throughout the body and the numbers could be adjusted for optimal rejuvenation. As desired the implants could be removed or replaced with better engineered cells as no permanent alteration is made to the mouse. By changing the type, placement, and number of implants various rejuvenation treatments could be tested alone or in combination. Local versus systemic effects could be studied.

This experimental platform could significantly accelerate the development of rejuvenation treatments that could then be immediately and safely tested in aged humans. No foreign drugs are introduced. The level of signalling molecules could be gradually increased by adding more implants. Implants could be quickly removed at the first sign of harm. The focus would not be on extension of maximum lifespan but rather be on restoration of youthful function in body tissues. So long trials would not be needed. The initial subjects would be very old so long term effects would be irrelevant as the subjects would naturally have died anyway. Hopefully an increase in maximum lifespan would be a welcome side effect.

Posted by: Anonymous at January 21st, 2016 2:32 PM

Oops...should have been Conboy, not Conway.

Posted by: Anonymous at January 21st, 2016 3:47 PM

I will ask our researcher Dr stolzing for her view on this.

Posted by: Steve h at January 21st, 2016 5:12 PM

The author is not aware that mesenchymal stem cells WITHOUT chemo have been in clinical trials for several years. Here is a recent interview with Dr. Sadiq, Tisch MS Center, announcing first ever repair and reversal of disability for progressed MS.
http://www.wheelchairkamikaze.com/2016/01/an-interview-with-dr-saud-sadiq-tisch.html?spref=fb

Those of us close to death who have run out of medical options are pioneering cellular therapy with good results, outside of clinical trials, because we've run out of time.

Would you call this rejuvenation due to stem cells?
Recent news story on recovery of two young patients who were terminal with Systemic Juvenile Idiopathic Arthritis, now healthy. The Arthritis Foundation is following closely, so hopefully a US study will happen.
http://www.fox26houston.com/health/healthworks/75037340-story

Rejuvenation of aging effects is a ho-hum side effect after you experience this kind of revival. Yes, wrinkles are less, skin problems gone, sex drive increases. We don't need to wait another decade for the perfect man-made designer induced pluripotent cells. Your own MSCs know what to do. They respond to cytokine signals to home in on injury and make repairs. Endogenous trophic factors trigger differentiation into the needed cell type.

Here is my report on using high dose MSC therapy to put secondary progressive MS into remission, published in Texas MD Magazine. And suggestions on accelerating trials and approval, to salvage the millions suffering from these god awful chronic and untreatable diseases.
http://www.texasmdmonthly.com/blog/cellular-therapies-to-the-rescue

Posted by: SammyJo at January 24th, 2016 10:04 PM

If you consider aging as a genetically programmed cascade process - beginning at sexual maturity, it would seem any treatment interventions focused only on specific aging symptoms (all current ones) that do not redirect the basic aging genetic program could only have specific and temporary effects until again over ridden. Essentially you can't meaningfully affect the aging process without altering the genetic cascade program that drives it. Obviously, that is no small feat.

Posted by: Durwood M. Dugger at January 27th, 2016 4:53 PM

I have an idea for you.
I'm aware that the chances that I've come up with something really new is absurdly low. So I'll be brief to avoid unduly wasting your time. OK here it is.
The big question about aging is: why does every living thing eventually age. Crosslinks, junk and so forth are good answers but potentially not complete answers, since it's possible at least in principle that evolution could find some way to eliminate those.
However, genetic mutation is something that evolution *cannot* fix except by reproduction, since for any given cell it's impossible to say whether the particular genes in it are the original ones or mutated ones, unless the cell does something crazy like go cancerous.
Humans, however, have the ability to determine the *exact* original genome, through statistical methods. We can culture a large population of cells and pick out the most common version of every gene. That is most likely the correct one. Evolution *can't* do that, which is why I think this is a plausible reason why all things age. The only thing evolution can do is reproduce a whole new body from a single cell every once in a while, and then have the new organism suffer death if there is a sufficiently bad mutation in it.
So the fix would be: culture one individual cell, and also culture a large group of cells. Run the dna of the large group and the individual separately. Then you can pick out every point mutation in the individual cell. Use CRISPR to fix every one of those mutations, and then induce those to become pluripotent stem cells, and the end result is not just stem cells but genetically *flawless* stem cells.
OK, that's it.

Posted by: Sean Carter at May 10th, 2016 12:12 PM

Rejuvenation or regeneration of stem cells is the simple path to health during long life. If senescent cells can be stimulated to return to a normal cell cycle then new healthy stem cells and organ cells will will be created. If new stem cells can be generated in the various niches then new organ cells will result. I am currently experiencing healthy body hair growth. My "dry mouth and dry eyes" appear to be alleviated. My nails are healthier:stiffer and thicker. My urine flow appears to have greater volume and pressure. I don't have a lab to test and prove what internal affects show stem cell rejuvenation but my subjective, and therefore irrelevant, opinion is there are many. If someone with a lab would contact me I would arrange to pay what little I have ( 74 and on Social security ) to defray costs of tests, to prove or disprove my opinion. If correct, this is obviously important. If not, determining the cause of the observable clinical conditions may well be important. Alleviating Sjorgen's syndrome would be important in itself.

Posted by: Roger Duronio at November 19th, 2016 10:06 AM

MSC and Dendritic are the way to go IMO

Immunologic T cell ……. teeter-totter*
"Ask Immunologic questions give molecular answers" Andy Blidy

11-12-2016
Andy Blidy

Regenerative Medicine cell replacement with Immunotherapy, Harnessing the Immune Systems to kill or promote life saving responses,

Why is the immune response like a big mountain peak? B/c if you fall off the mountain top b/c your immune system is damaged/ aged do to many factors some environmental and some genetically…..This can result in disease state of aging and you can get an auto immune disease on one side of the mountain or cancer on the other side of the mountain….. But how to climb back to the top of the immune mountain with a replacement cellular genetic expression that results in homeostasis of the immune system ? …Balance is the key in immunology, tolerance control by both the MHC1 and MHC2 pathways….If you are "health" or young your gamma delta /alpha beta cells will keep you homeostasis or balance at the top of the mountain. It is now known that gamma delta show suppression of the immune system while alpha beta are the fulcrum for proliferation during an immune response or in attack mode. Knowing the cellular process and their checkpoints are the key.

1. Immunotherapy Checkpoint
"Antigen-presenting cells (APCs) are a heterogeneous group of immune cells that mediate the cellular immune response by processing and presenting antigens for recognition by certain lymphocytes such as T cells CD4 helper cells and CD8 cytotoxic with the furculum at the TCR T cell receptor which have two alpha beta units to one gamma delta structure. Classical APCs include dendritic cells, mesenchymal and epithelial (EMT), macrophages, monocytes Langerhans cells and B cells". This is why antigen presentation is key to treating a trauma cell repair, cancer or autoimmune diseases at first line of defense to the individual? These cells present the T cell with an antigen from say a cancer cell with a co- stimulator protein…….This process begins to hunt down diseases by NK cells and CD8 killers cells that are covered with the same antigens or the repair by CD 4 cells.

2. Immunotherapy Checkpoint
Activated T cells "A T cell or T lymphocyte is a type of lymphocyte (a subtype of white blood cell) that plays a central role in cell-mediated immunity. T cells can be distinguished from other lymphocytes, such as B cells and natural killer cells, by the presence of a T-cell receptor on the cell surface." Once activated you will alter the microbiome to respond to cancers or any foreign antigen or suppress the immune system in normal functions, the cause of auto immune disease such as Parkinson , MS, lupus. Key subset is Tregs CD4 CD25 FOXp3, GARP, LAP , SATB! cells and the subsetting of all TH1, TH2 and TH17. Foxp3 is shown to be the master regulator of Tregs at the transcription level with functional gene regulation by SATB1(special AT-rich binding protein), LAP (latency-associated peptide),GARP that tethers to TGF-beta …….This is the marriage of molecular biology with immunology at the cell surface and gene expression levels…… How does EMT function as a microbiome translation with ACT T cells ? Same is true for CD8 activated cell but less study over the years B/C of sub populations .

3. Immunotherapy Checkpoint
Alpha beta gamma delta T cells what are they? "The seesaw (also known as a teeter-totter or teeterboard) is a long, narrow board supported by a single pivot point, most commonly located at the midpoint between both "ends"; as one end goes up, the other goes down." In the middle immune system at the fulcrum are either your gamma delta or alpha beta expression.

Gamma delta from the TCR
Figure 1
"Ligand recognition by αβ and γδ TCRs.
• Full size figure and legend
"("a) The αβ T cells recognize peptide bound to MHC molecules through the interaction of both TCR chains with peptide-bound MHC molecules. β2m, β2-microglobulin. (b) In contrast, the NKT cell TCR contacts CD1d-bound glycolipid through its invariant TCRα chain. (c,d) The recognition of ligand by γδ T cells is less well understood. (c) The G8 γδ TCR inserts its CDR3δ loop into the hydrophobic groove of T22, with the γ-chain making only a minor contribution to binding. (d) Willcox et al. now show a unique mode of binding for another γδ TCR, LES, to its ligand, EPCR/ The CDR3 loop of the LES TCR γ-chain binds to EPCR independently of lipid antigens through residues on the EPCR β-sheet."

EPCR: a stress trigger for γδ T cells
• Deborah A Witherden
• & Wendy L Havran
Nature Immunology13, 812-814(2012)
doi:10.1038/ni.2398
Published online
21 ugust 2012

4. Immunotherapy Checkpoint
CTLA-4 Protein what is its function? "CTLA-4 (cytotoxic T-lymphocyte-associated protein 4), also known as CD152 (cluster of differentiation 152), is a protein receptor that, functioning as an immune checkpoint, down regulates immune responses." The protein is the brakes of the cellular immune response. …..cancer cell immune checkpoints that shut down T cell response and allow the cancer stem cells to grow undisturbed…..Checkpoint inhibitors will physically block the checkpoint , which free Act T and other T cells to kill cancer cells.

5. Immunotherapy Checkpoint
Last line of defense of self recognizing of good vs. bad before apoptosis cell death …… "PD-1 or Program death of the immune cells CD279 (cluster of differentiation 279), is a protein that in humans is encoded by the PDCD1 gene. "PD-1, functioning as an immune checkpoint, plays an important role in down regulating the immune system by preventing the activation of T-cells, which in turn reduces autoimmunity and promotes self-tolerance. The inhibitory effect of PD-1 is accomplished through a dual mechanism of promoting apoptosis (programmed cell death) in antigen specific T-cells in lymph nodes while simultaneously reducing apoptosis in regulatory T cells(suppressor T cells"
This is the MAB that saved Jimmy Carter's life, KeyTrude

http://www.nbcnews.com/health/cancer/cancer-drug-keytruda-keeps-some-patients-alive-3-years-n576376

5a. Immunotherapy Checkpoint
One way is the IDO-1 "This gene encodes indoleamine 2,3-dioxygenase (IDO) This enzyme is thought to play a role in a variety of pathophysiological processes such as antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Through its expression in dendritic cells, monocytes, and macrophages this enzyme modulates T-cell behavior by its peri-cellular catabolization of the essential amino acid tryptophan . IDO is an immune checkpoint molecule in the sense that it is an immunomodulatory enzyme produced by some alternatively activated macrophages and other immunoregulatory cells (also used as an immune subversion strategy by many tumors). Interferon-gamma has an antiproliferative effect on many tumor cells and inhibits intracellular pathogens such as Toxoplasma and Chlamydia, at least partly because of the induction of indoleamine 2,3-dioxygenase" causing depletion of tryptophan which can cause halted growth of microbes as well as T cells.[7] PGE2 is able to elevate the expression of indoleamine 2,3-dioxygenase in CD11C(+) dendritic cells and promotes the development of functional Treg cells"

This IDO-1 pathway MUST be combine with say PD-1 protein blocking pathway or other cellular s such as mesenchymal or plasmacytoid dendritic cells (pDCs) and antigen presentation to call for more killer cells.

What controls the down regulation of the immune system at the microbiome Epithelial to Mesenchymal Cell Transition (EMT) and therefore sends the human body in the reboot, rescue and repair modes by helper CD4 T cells ? Mesenchymal cells

"Epithelial to Mesenchymal Cell Transition - loss of cell adhesion leads to constriction and extrusion of newly mesenchymal cell."

How about the proliferation of say TIL (Tumor infiltrated lymphocytes ) cells get exhausted ? "Lack of Dendritic cells maybe at the microbiome effecting the dendritic cells are usually divided into two main groups: the myeloid dendritic cells (mDCs) and the plasmacytoid dendritic cells (pDCs) and antigen presentation to call for more killer cells."

Full steam ahead with this Immunotherapy treatment we are innovating a new paradigm shift with stem cells at the Mesenchymal and dendritic levels with new cells such as hESCs/IPSC or simple new cell replacement be it gene therapy . Early the cell the better and they migrated to the site of need to alter the microbiome environment.

We need to complete studies ASAP to increase demand of immune therapy and cellular response. We have started the development phase of setting up the manufacturing process for release of an approved cellular products. When I started this thread, I was thinking what is a way to short cut the drug product approval cycle, since this is regenerative medicine and we are using cells to treat and halt these diseases? We are trying to release a product for the #1 unmet medical need with a new technology with limited resources (conservative money, no JV yet and no revenues) and the FDA/NIH and world regulatory bodies are on a learning curve.

The patient's needs are for treatment, yesterday, time lost/ life lost. Who would not want to improve their eye sight with a safe effective treatment or cure cancer like Jim Allison with his CTLA-4 Monoclonal antibodies or using new replacement cells to save lives of your love ones?

My questions are when will the patients' unmet needs Trump the need of process? And where can the drug process be short circuit for the benefits of the patients? Federal FDA/NIH funding or CIRM funding in California.

After all folks we are tied to our patient's diagnosis, prognosis, and cellular treatment of the FDA in the USA. Can other countries such as Japan move this process more quickly since they are more focused on the immune process? Time will tell……In the meantime many will suffer and die today…..

Part 2

How does cell replacement / Regenerative medicine work with the immune system? What happens when you inject stem cells that can migrate to site of need? How do you define potency of cells and protein cytokines effects on Microbiome? Do you know , let's talk Think about what a cancer stem cell is and early immune monitoring is the key for potency, migration and paracrine cytokines . Where scientist are mislead is they deal in the adult system without regards of cell replacement with the correct state …….If you have stage IV cancer you beeter understand PD-1 biology????

Three key terms antigen presentation (be it protein or cell-cell interaction) potency (naive blastomere pluripotent stem cells are the gold standard) and migration (or in cancer know as metastasis) and a possible answer to a question of Cancer Stem Cells and what are they? You are more that welcome to share this white paper … Why are the T cells and NK cells considered the police and fire departments? And is Cancer the burning building ? Who do you call when you have a burning building? The plumber ( T cells )!!!!!!!! Can T And Natural Killer (NK) cells kill cancer? Answer is yes we have know that for 30 years ....Lewis L. Lanier ( NK s) and Jim Allison ( CTLA-4) ….What controls T cells at the microbiome ( site of the cancer )? MSC cells, down regulation !!!!!What controls NK cells at the microbiome? Dendritic cells, up regulation !!!!!! more on the up and down regulation of the immune system with these cell types?

Highly tumorigenic subpopulation of cancer cells expressing or lacking the cell surface markers CD10, CD24, CD44, CD133. Stro-1 and epithelial-specific antigen (ESA) CAMs are the key to understanding the immune of Cancer. You can substitute any CAM for any organ to be more comprehensive or inclusive tumor specific ……

Why is CD24 key to mesenchymal stem cells …..Why are they very high in blastomere pluripotent stem cells 24% vs. adult MSC 0-1 % ….Lanza MSC papers on Ocata.com

1. What is CD 24 "Signal transducer CD24 also known as cluster of differentiation 24 or heat stable antigen CD24 (HSA) is a protein that in humans is encoded by the CD24gene.[1] CD24 is a cell adhesion molecule." Very important and "CD24 is a glycoprotein expressed at the surface of most B lymphocytes and differentiating neuroblasts. This gene encodes a sialoglycoprotein that is expressed on mature granulocytes and in many B cells. " And we now know on early embryonic MSC from blastomere pluripotent stem cells -Lanza MSC papers

2. What is CD44 homing receptor know a lot about this ….. "CD44 antigen is a cell-surface glycoprotein involved in cell-cell interactions, cell adhesion and migration. In humans, the CD44 antigen is encoded by the CD44 gene on Chromosome 11.[1] CD44 has been referred to as HCAM (homing cell adhesion molecule), Pgp-1 (phagocyticglycoprotein-1), Hermes antigen, lymphocyte homing receptor, ECM-III, and HUTCH-1." what is metastases you got it ….migration

3. CD10 this is the blast state of the immature cells "Neprilysin, also known as membrane metallo-endopeptidase (MME), neutral endopeptidase (NEP),cluster of differentiation 10 (CD10), and common acute lymphoblastic leukemia antigen (CALLA) is an enzyme that in humans is encoded by the MME gene. Neprilysin is a zinc-dependent metalloprotease that cleaves peptides at the amino side of hydrophobic residues and inactivates several peptide hormones including glucagon, enkephalins, substance P, neurotensin, oxytocin, and bradykinin.[1] It also degrades the amyloid beta peptide whose abnormal misfolding and aggregation in neural tissue has been implicated as a cause of Alzheimer's disease." And we now know on early eMSC from blastomere pluripotent stem cells -Lanza MSC papers

4. Stro-1 What is Stro-1? " Stro-1 is the best-known mesenchymal stem cell marker. However, despite its bone marrow origin, its localization in bone marrow has never been demonstrated. By immunofluorescence staining, it is shown that ∼ 0.74% of nucleated bone marrow cells expressed Stro-1. Also found that ∼ 8.7% of CD34-expressing cells expressed Stro-1, and more than 20% of Stro-1-expressing cells did not express CD34. In adipose tissue Stro-1 expression was identified in the endothelium of arterioles and capillaries. Stro-1 was also localized in the endothelium of some but not all adipose tissue veins. Endothelial expression of Stro-1 was also identified in blood vessels in penis and in leg muscles, but not in other tested tissues. In these other tissues, Stro-1 was scantly expressed near but not in blood vessels. These variable and endothelial expression patterns of Stro-1 point to a need to re-examine published data that relied on Stro-1 as a mesenchymal stem cell marker."

Lanza found that Stro-1 was negative for his hMSC blastomere pluripotent stem cells vs. very positive for adult MSC ….so if you have Stro-1 in high expression your potency will be varied and low…An err of many MSC researchers because of the maturity of MSC from EMT…..

5. "ESA Or CD326 Ep-CAM consists of two glycoproteins, 34 and 39 kDa, sometimes designated epithelial antigen, epithelial specific antigen, and epithelial glycoprotein. In paraffin sections, the protein is detected with mAbs like Ber-EP4 and MOC-31. The glycoproteins are located on the cell membrane surface and in the cytoplasm of virtually all epithelial cells with the exception of most squamous epithelia, hepatocytes, renal proximal tubular cells, gastric parietal cells and myoepithelial cells."

6. CD133 "CD133 antigen also known as prominin-1 is a glycoprotein that in humans is encoded by the PROM1gene.[1][2] It is a member of pentaspan transmembrane glycoproteins (5-transmembrane, 5-TM), which specifically localize to cellular protrusions. While the precise function of CD133 remains unknown, it has been proposed to act as an organizer of cell membrane topology" "Tissue distribution[edit]

CD133 is expressed in hematopoietic stem cells,[4] endothelial progenitor cells,[5] glioblastoma, neuronal andglial stem cells,[6] various pediatric brain tumors,[7] as well as adult kidney, mammary glands, trachea, salivary glands, placenta, digestive tract, testes, and some other cell types.[8][9]"

7. All these CAM reagents are important because of EMT

"Epithelial to Mesenchymal Transition (EMT) describes a mechanism by which cells lose their epithelial characteristics and acquire more migratory mesenchymal properties. This transient and reversible process is classified into three subtypes that are dependent on the biological and functional setting in which it occurs. This illustration represents general pathways in the scientific literature and is not to be considered comprehensive nor definitive. TIL tumor-infiltrating-lymphocytes Transcriptomes of T helper 1 (Th1), Th17. Treg cells have early cell surface markers such as programmed death (PD)-1 and CCR8. Therefore major histocompatibility complex (MHC) molecules play a major function in immunotherapy.

Type 1 EMT during development is essential for gastrulation, neural crest cell migration, and organ development. EMT generates fibroblasts following tissue injury that assist in local wound healing.

Type 2 Persistent EMT following attenuation of inflammation can result in organ fibrosis.

Type 3 EMT results in the transformation of epithelial cells into the invasive metastatic mesenchymal cells that underlie cancer progression."

"Loss of Tight Junctions, Adherens Junctions, and Desmosomes > Disassembly of specialized cell-cell contacts leads to redistribution of cytoskeletal proteins and disruption of the apical-basal cell polarity of epithelial cells. > Key Molecules: Actin, α-Actinin, α-Catenin, β-Catenin, Claudins, E-Cadherin, Desmogleins, Desmocollin, JAM, Occludin, Plakoglobin, Plakophilin, Vinculin, Zona Occludens

Cytoskeletal Changes Formation of actin stress fibers that anchor to focal adhesion complexes to begin to promote cell migration. > Key Molecules: Actin, Cytokeratins, S100A4, α-Smooth Muscle Actin, Vimentin

Transcriptional Shift Suppression of epithelial genes and activation of mesenchymal genes is mediated by Snail, ZEB, and bHLH family transcription factors. Vimentin is upregulated and extracellular deposition of Fibronectin is increased. > Key Molecules: FoxC2, Goosecoid, LEF-1, Snail 1, Snail 2 (Slug), Twist-1, ZEB1, ZEB2

Increased Migration and Motility > Up regulation of N-Cadherin, secretion of matrix metalloproteases, and stimulation of integrins by extracellular matrix proteins facilitates cell motility. > Key Molecules: N-Cadherin, FAK, Fibronectin, α5β6 Integrin, Laminin-5, SPARC, Syndecan-1, Vitronectin these are your cancer causing molecules"

Embryonic DC cells ..Why? …..homogeneous population easy of manufacturing and they are the most potent …..Do they migrate like hMSC mesenchymal cells? ….unknown

Take in concretion the dendritic cells What are DC ? "Dendritic cells (DCs) are antigen-presenting cells (also known as accessory cells) of the mammalian immune system. Their main function is to process antigen material and present it on the cell surface to the T cells of the immune system."
And "The innate immune system is an evolutionarily older defense strategy, and is the dominant immune system found in plants,fungi, insects, and primitive multicellular organisms.[3]
The major functions of the vertebrate innate immune system include:
• Recruiting immune cells to sites of infection, through the production of chemical factors, including specialized chemical mediators, called cytokines
• Activation of the complement cascade to identify bacteria, activate cells, and promote clearance of antibody complexes or dead cells"
• The identification and removal of foreign substances present in organs, tissues, the blood and lymph, by specialised white blood cells
• Activation of the adaptive immune system through a process known as antigen presentation"
Acting as a physical and chemical barrier to infectious agents. "Dendritic cells (DC) are phagocytic cells present in tissues that are in contact with the external environment, mainly the skin (where they are often called Langerhans cells), and the inner mucosal lining of the nose, lungs, stomach, and intestines.[2] They are named for their resemblance to neuronal dendrites, but dendritic cells are not connected to the nervous system. Dendritic cells

Posted by: Andy Blidy at November 25th, 2016 4:43 AM

part # 2

Mesenchymal, Dendritic Cells Immunologic Cell Teeter-Totter Controlling the Immune System
"Ask Immunologic questions give molecular answers" Andy Blidy
https://www.youtube.com/watch?v=PphTL6vY4W4 History of Immunotherapy by James Allison at PMWC 2017 Silicon Valley

https://www.youtube.com/watch?v=yMxegkW9p-k Lewis L. Lanier Immunotherapy: Unleashing the Body's Natural Defense Systems to Fight Cancer

Version 1.1
8-5-2017

Why is the immune response like a Teeter Totter? B/c if you fall off the Teeter Totter b/c your immune system is damaged/ aged due to many factors, some environmental and some genetically, you end up in a disease state…..This can result in disease state of aging and you can get an auto immune disease on the bottom end of the immunologic Teeter Totter or cancer on the top side of the Teeter Totter….. But how does one balance the immune Teeter Totter as one age's a replacement cellular therapy will be employed in the near future and genetic expression from possible gene therapy ( CRISPR-CAS9 technology) that results in homeostasis of the immune system? …Balance is the key in immunology, tolerance control by both the (Major Histocompatibility Complex) MHC I and MHC II pathways….If you are "healthy" or young, your gamma delta /alpha beta cells will keep you homeostasis or balance on the Teeter Totter. Think of certain cells as the fulcrum on the Teeter Totter. A question can be asked, Do you want to suppress the immune system or activate it? But what cells control the microenvironment to do this? A high level process of the immune system is illustrated below in several illustration ……Notice balance is needed in these quantum immunologic systems with the Mesenchymal (MSC) and Dendritic cells (DC) in the middle of balance immunity acting as fulcrum points of checkpoints associated with biomarkers …….Now let's look at the microbiome and the immunomodulatory effects of Mesenchymal and Dendritic cells at the local tissue level related to immune system and how these systems function together, keeping in mind the effects on auto immune diseases and cancers in an inflammation or tumor microenvironment or tissue organ disease….

Mesenchymal / Pericytes are contractile cells that wrap around the endothelial cells. What is epithelial transition to mesenchymal, EMT? Answer is in last illustration….(MSC) are near such tissue as the skin and mucous membranes, providing the first line of defense for innate immunity. Many non-myeloid cells contribute to this defense strategy, including fibroblasts and epithelial/mesenchymal cells, producing immunomodulatory and antimicrobial factors. MSCs and IMO are centrally located within the epithelial microbiome associated with both of the innate cellular CD8 and humor B cell of immune system. Expression of toll-like ligand receptors (TLRs) on the cell surface of mesenchymal stromal cells (MSCs) is another key role suggesting their inherent role in recognition of antigen presentation

MSCs effects many different cells and are a major component of the innate immunity trigging the cascade of the complement system. The central step in complement activation is the cleavage of C3 into C3a and C3b. These proteins become bound to the immune cells and express the complement receptors, such as CR1, CR2, CR3, and CR4. Binding of MSC to C3 has been shown to suppress the proliferation of Peripheral Blood Mononuclear Cells. Therefore, this inhibits Activated T cells that are required for rescue and repair by CD4 pathways. This suppressive activity of MSCs may be a major role to regulating immune control of the regeneration and repair of tissue. Notice the Mesenchymal MSC is the central cell of this biological system …..

"The multi-faceted anti-inflammatory actions of MSCs. In response to pro-inflammatory cytokines or TLR3 stimuli, MSCs will develop an anti-inflammatory profile. Through the secretion of soluble factors these licensed cells can act on numerous innate immune cells affecting both effector function and phenotype. Individual effects are discussed further within the main text.https://doi.org/10.1016/j.imlet.2015.05.004 Mesenchymal stromal cells and the innate immune response" reference #1

Other cells affected by MSCs are:
1. Neutrophils (PMNs) are major part of innate immune cell, responding to foreign challenge by homing to the wound site within several minutes of injury. These non-proliferative, phagocytic cells respond to such foreign antigen as bacterial invasion, these are control in the microbiome by MSCs such as in sepsis to help in microbial defense. Myeloid-derived suppressor cells (MDSCs) implicated in Autoimmune Diseases and Cancer to maintain a cancer microenvironment or chronic inflammation microbiome.
The immunology and molecular biology has come a long ways from the Aids Crisis in the early 1980's to the embryonic age in 1998 and the field has not developed a bio-process manufacturing of MSC cell in large volume for therapy by cell replacement. In fact the MSC are non-standard for potency, migration and Paracrine production.

2. Mast cells (MCs) are also innate responder cells during MSC homing levels of IL-6 IL-1a secretions aid in the prevention of pro-apoptotic activity on neutrophils. MCs are related to IgE mediated MC degranulation that can call for the recruitment of PMNs a Dendritic cells. Most scientists relate MCs to allergy pathways. Including myself.

3. Dendritic Cells (DCs) provide a link between the innate and adaptive immune systems ( Cartoon below)

"Immunosuppression by mesenchymal stem cells. MSCs suppress innate and adaptive immune responses by enhancing regulatory immune cells with tolerogenic properties. MSCs suppress macrophages by favoring monocyte polarization to anti-inflammatory M2 macrophages, increasing the production of IL-10, and decreasing the production TNF-α and IL-12. MSCs can also regulate DCs by downregulating the expression of MHC, CD40, CD80, CD83 and CD86, thus, diminishing their antigen presenting ability, while upregulating the expression of IL-10. MSCs can reduce the NK cell cytotoxicity and decrease their production of TNF-α and IFN-γ. Treg and Breg cells can be induced by MSCs, further increase the production of anti-inflammatory cytokines (IL-10 and TGF-β1). However, the mechanisms of how Breg cells are induced by MSCs are still not clear. MSCs: Mesenchymal stem cells; TNF: Tumor necrosis factor; IL: Interleukin; NK: Natural killer; DCs: Dendritic cells; IFN-γ: Interferon-γ; Treg: Regulatory T; Breg: Regulatory B; TGF: Transforming growth factor; PGE2: Prostaglandin E2; IDO: Indoleamine 2,3-dioxygenase."
World J Stem Cells. 2016 Sep 26; 8(9): 268-278.
Published online 2016 Sep 26. Doi: 10.4252/wjsc.v8.i9.268

4. Natural killer (NK) cells have are activated by exposure to IL-2 or IL-15 Causing the expression of IFNγ and TNFα pathways for cancer tumor and viral infection….. Both IDO and PGE2 offer multiple mechanisms for dampening NK responsiveness to the MSCs.

5. Monocytes represent approximately 10% of circulating leukocytes detecting microbial pathogens and enhance the recruitment of monocytes and macrophages into inflamed tissues to promote wound repair through the secretion of the chemokine.

6. Macrophages are characterized into two phenotypes, the M1 pro-inflammatory macrophage with antimicrobial activity and the M2 anti-inflammatory macrophage

What is the allogenic effect of MSC on the response related to immune T cells? In vivo Robert Lanza and group have shown that human embryonic MSC does alter the secretion of cytokines to Dendritic Cells, Naive and effector T cells both TH1 and TH2 and NK s causing an anti-inflammatory response along with phenotype change. tumor necrosis factor (TNF- alpha) secretion and mature DC to increase interleukin-10 (IL-10) and interferon gamma (IFN-g) this will cause an increase in Treg CD4 cells by IL-4 …Leading to prostaglandin E2 (PGE2), and inhibitors of PGE2 production mitigated hMSC-mediated immune modulation in the microenvironment that controls inflammation and may lead to lower GVHD disease from transplantation of cells or organs. Recently, McGuirk (Reference #3 ) has shown MSCs secrete an array of cytokines, chemokines, and soluble receptors that act locally in microbiome. MSCs regulate immunity by interacting with innate immune cells (including macrophages, natural killer (NK) cells, and dendritic cells), and adaptive immune cells (including B and T cells)

"Immune profile of mesenchymal stem cells (MSCs). Graphic summary of the interactions between MSC and the immune system. MSCs can suppress proliferation of both T helper (TH) and cytotoxic T cells (Tc) through multiple pathways. Differentiation of MSCs to TH2 and regulatory T-cells (Treg) is triggered, resulting in an anti-inflammatory environment. Interleukin (IL)-6 blocks the maturation of dendritic cells (DC) by inhibiting upregulation of CD40, CD80, and CD86, which subsequently reduces T-cell activation. Monocytes are stimulated by MSCs to preferentially differentiate towards the M2 phenotype. IL-10, produced by M2 macrophages, can boost the formation of Treg, and simultaneously reduces neutrophil tissue migration. Neutrophils (polymorphonuclear granulocytes; PMN) have a longer life span; however, production of reactive oxygen species (ROS) is decreased. Natural killer (NK) cell proliferation and cytotoxic activity are both suppressed. B-cell proliferation is inhibited, and production of antibodies is reduced. HGF, hepatocyte growth factor; IDO, indoleamine-pyrrole-2-3-dioxygenase; PGE2, prostaglandin E2; and TGF-β, transforming growth factor-β. (Adapted from van den Akker F, de Jager SC, Sluijter JP. Mesenchymal stem cell therapy for cardiac inflammation: iummunomodulatory properties and the influence of toll-like receptors." Mediators Inflamm2013: 181020, 2013.)

"Mesenchymal stromal cells (MSCs) are adult multipotent stem cells residing as pericytes in various tissues and organs where they can differentiate into specialized cells to replace dying cells and damaged tissues. These cells are commonly found at injury sites and in tumors that are known to behave like "wounds that do not heal." the mechanisms of MSCs in migrating, homing, and repairing injured tissues. Reference also reviews a number of reports showing that tumor microenvironment triggers plasticity mechanisms in MSCs to induce malignant neoplastic tissue formation, maintenance, and chemoresistance, as well as tumor growth" Reference #5

"MSC role in tumor microenvironment. MSCs can induce cancer cell survival, stemness, and chemoresistance by differentiating
into cancer-associated fibroblasts (CAFs) using a tumor growth factor β type 1 (TGFβ1)-dependent mechanism, and by releasing soluble
factors that favor angiogenesis and immunosuppression in the tumor microenvironment, such as prostaglandin E2 (PGE2) and vascular
endothelial growth factor (VEGF). MSCs can mediate anti-cancer effects by releasing anti-cancer factors, such as tumor necrosis factor related
apoptosis inducing ligand (TRAIL), via mechanisms that are not well understood."
Cancer Biol Med 2017. doi: 10.20892/j.issn.2095-3941.2016.0033

In Conclusion:
It is now known that TCR gamma delta show suppression of the immune system while TCR alpha beta is the fulcrum for proliferation during an immune response or in attack mode. Knowing the cellular process and their checkpoints are the key. I have outlined 5 major checkpoints on the immune Teeter Totter as:

1.) Antigen presentation, start of the innate immunity system
2.) Activated T Cells, the aggressive protection expansion of the human immune system
3.) T Cell receptor, the integrators that discriminate self vs. non self
4.) "The CTLA-4 (cytotoxic T-lymphocyte-associated protein the brakes of the immune response and finally
5.) PD-1 (Programmed death -1) killing of foreign invaders or aging cells or cancerous cells of apoptotic gone wrong
An example of Tumor resistance that may be alter by the correct MSC cell population. Indoleamine 2,3-dioxygenase (IDO) is the rate-limiting enzyme in the catalysis of tryptophan. The mechanism by which IDO promotes tolerance is still an area of active investigation and hot clinical trials for Cancer with PDL-1 and other biomarkers, such as CTLA-4.

Immune checkpoint pathways down regulate or up regulation of T-cell activation to maintain peripheral tolerance can be exploited by tumors to induce an immunosuppressive state that allows the tumors to grow and develop instead of being eliminated by the immune system. The differential patterns of the CTLA-4 and PD-1 ligand expression-found primarily in lymphoid tissue and in peripheral tissues, respectively-are central to the hypothesis that CTLA-4 acts early in tolerance induction and PD-1 acts late to maintain long-term tolerance. One needs to reboot the immune with the correct stem cells this paper will outline the repair of chronic inflammation or cancer …….The immune system needs to be rebooted so rescue and repair can take place. But MSCs will be the key in stem cell replacement in Regenerative Medicine in the near future. The major question is what MSC type of Replacement cell, adult vs embryonic vs IPSC ? IMO, Naïve blastomeric pluripotent MSC ( google Robert Lanza and MSC) derived from NED cell are the most potent, migrate the best and produce the immunomodulatories need for reboot, rescue and repair, therefore will be the best cells for therapy , but how long will the bioprocess take and the scientific field to realize what are the best cells ?

Major problem of Tumor infiltrated Lymphocytes, TIL Cells in Cancer……What is the solution? Can MSC control fibroblast and be the magic cell to help cure cancer? IMO, yes

Learning from PD-1 Resistance: New Combinations Strategy Gordon J. Freeman

References:

1. Mesenchymal stromal cells and the innate immune response, Volume 168, Issue 2, December 2015, Pages 140-146, Immunology letters Katarina Le Banc and Linsay C. Davies
2. Human mesenchymal stem cells modulate allogeneic immune cell response Blood 2005 ,105 1815 -1822 Sudeepta Aggarwal and Mark F. Pittenger
3. Review: Mesenchymal Stromal Cells: What Is the Mechanism in Acute Graft-Versus-Host Disease? Neil Dunavin , Ajoy Dias , Meizhang Li and Joseph McGuirk , biomedicine 1 July 2017 5, 39 doi:10,3390
4. Rebuilding the Damaged Heart: Mesenchymal Stem Cells, Cell-Based Therapy, and Engineered Heart Tissue Samuel Golpanian, Ariel Wolf, Konstantinos E. Hatzistergos, Joshua M. Hare Physiological Reviews Published 22 June 2016 Vol. 96 no. 3, 1127-1168 DOI: 10.1152/physrev.00019.2015
5. Mesenchymal stromal cells' role in tumor microenvironment: involvement of signaling pathways Armel Herve Nwabo Kamdje1, Paul Takam Kamga2, Richard Tagne Simo1, Lorella Vecchio3, Paul Faustin Seke Etet3, Jean Marc Muller4, Giulio Bassi2, Erique Lukong5, Raghuveera Kumar Goel5, Jeremie Mbo Amvene1, Mauro Krampera2 Cancer Biol Med. 2017 May; 14(2): 129-141. doi: 10.20892/j.issn.2095-3941.2016.0033

Posted by: Andrew Blidy at August 25th, 2017 3:23 PM

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