Turning Pluriptoent Human Embryonic Stem Cells into a Large Supply of Plastic CNS Derivatives for Cell Therapy

San Diego Regenerative Medicine Institute and Xcelthera announce the publication of Dr. Parsons’ original research article supported by the National Institutes of Health, titled “An Engraftable Human Embryonic Stem Cell Neuronal Lineage-Specific Derivative Retains Embryonic Chromatin Plasticity for Scale-Up CNS Regeneration”, in Journal of Regenerative Medicine & Tissue Engineering.

Human stem cell transplantation represents a promising therapeutic approach closest to provide a cure to restore the lost nerve tissue and function for a wide range of devastating and untreatable neurological disorders. However, to date, lack of a clinically-suitable source of engraftable human stem/progenitor cells with adequate neurogenic potential has been the major setback in developing effective cell-based therapy as a treatment option for restoring the damaged or lost central nervous system (CNS) structure and circuitry. The traditional sources of engraftable human stem cells with neural potential for transplantation therapies have been multipotent human neural stem cells (hNSCs) isolated directly from the human fetal CNS. However, cell therapy based on CNS tissue-derived hNSCs has encountered supply restriction and difficulty to use in the clinical setting due to their limited expansion ability and declining plasticity with aging, potentially restricting the tissue-derived hNSC as an adequate source for graft material.

Alternatively, the pluripotent human embryonic stem cells (hESCs) proffer cures for a wide range of neurological disorders by supplying the diversity of human neuronal cell types in the developing CNS for regeneration and repair. We must bear in mind that the pluripotent hESC itself cannot be used for therapeutic applications. It has been recognized that pluripotent hESCs must be transformed into fate-restricted derivatives before use for cell therapy. However, realizing the therapeutic potential of hESC derivatives has been hindered by the current state of the art for generating functional cells through multi-lineage differentiation of pluripotent cells, which is uncontrollable, inefficient, instable, highly variable, difficult to reproduce and scale-up, and often causes phenotypic heterogeneity and instability, hence, a high risk of tumorigenicity following transplantation. Under protocols presently employed in the field, the prototypical neuroepithelial-like nestin-positive hNSCs, either isolated from CNS in vivo or derived from pluripotent cells in vitro via conventional multi-lineage differentiation, appear to exert their therapeutic effects primarily by their non-neuronal progenies through producing trophic and/or neuroprotective molecules to rescue endogenous dying host neurons, but not related to regeneration from the graft or host remyelination.

We recently reported that pluripotent hESCs maintained under a defined platform can be uniformly converted into a cardiac or neural lineage by small molecule induction. This technology breakthrough enables well-controlled generation of a large supply of neuronal lineage-specific derivatives across the spectrum of developmental stages direct from the pluripotent state of hESCs with small molecule induction. Having achieved uniformly conversion of pluripotent hESCs to a neuronal lineage, in this study, the expression and intracellular distribution patterns of a set of chromatin modifiers in the hESC neuronal lineage-specific derivative hESC-I hNuPs were examined and compared to the two prototypical neuroepithelial-like hNSCs either derived from hESCs in vitro or isolated directly from the human fetal CNS in vivo. These hESC-I hNuPs expressed high levels of active chromatin modifiers, including acetylated histone H3 and H4, HDAC1, Brg-1, and hSNF2H, retaining an embryonic acetylated globally active chromatin state. Consistent with this observation, several repressive chromatin remodeling factors regulating histone H3K9 methylation, including SIRT1, SUV39H1, and Brm, were inactive in hESC-I hNuPs. These Nurr1-positive hESC-I hNuPs, which did not express the canonical hNSC markers, yielded neurons efficiently (> 90%) and exclusively, as they did not differentiate into glial cells, such as astrocytes, and oligodendrocytes. Following engraftment in the brain, hESC-I hNuPs yielded well-dispersed and well-integrated human neurons at a high prevalence. No graft overgrowth, formation of teratomas or neoplasms, or appearance of non-neuronal cell types was observed following engraftment. Transplanted wild type mice developed hyper-active behavior, such as fast movement and fast spin, which also suggested that transplanted human neuronal cells had survived and integrated into the mouse brain to function and control mouse behavior. By contrast, the prototypical neuroepithelial-like nestin-positive hNSCs derived either from hESCs or CNS can spontaneously differentiate into a mixed population of cells containing undifferentiated hNSCs, neurons (10-30%), astrocytes, and oligodendrocytes in vitro and in vivo. These observations suggest that, unlike the prototypical neuroepithelial-like nestin-positive hNSCs, these in vitro neuroectoderm-derived Nurr1-positive hESC-I hNuPs are a more neuronal lineage-specific and plastic human stem cell derivative, providing an engraftable human embryonic neuronal progenitor in high purity and large supply with adequate neurogenic potential for scale-up CNS regeneration as stem cell therapy to be translated to patients in clinical trials.  

Controlled by Financial Conflicts --- How Did CIRM Board Members Preempt Prop 71 Bond? Why Would Stem Cell Meetings Sponsored by CIRM Blacklist Human Embryonic Stem Cell Research?

Financial conflict of interest without transparency is the formula for abuse and government corruption that CIRM (California Institute for Regenerative Medicine), the California state stem cell agency, has. Anyone with slightest scientific integrity would be shocked to see ICOC such daring undertaken by financial conflicts on California stem cell report, not just a few millions, it is billions, it is hundreds of millions of government fund disappearing into CIRM board’s own institutions and companies without asking any questions if they have any scientific merits and if they have followed the legal procedure of Prop 71. How come Alan Trounson dared to claim that there is no evidence of financial conflicts of interest for CIRM awards on California stem cell report. Maybe Alan Trounson is at easy to know that he and CIRM have already set up many non-transparent safety precautions to cover up any evidences, e.g. use pre-application to deliberately select conflict of interest awards, never disclose any information of those pre-applications, and never allow investigator-initiated applications and appeals for pre-applications. Maybe Alan Trounson is at easy to know that he and CIRM have already preempted Prop 71 bond prior to ICOC public meetings, that no matter which award would come up, no matter who would appeal and win the appeal, the money would always go to CIRM board members’ institutions. Alan Trounson and CIRM have intentionally given California stem cell researchers many difficulties to even apply for CIRM funding by denying them the rights to apply for public fund by law, such as non-transparent pre-application procedures (see our previous posts) and conflict of interests RFAs (e.g., CIRM RFA 12-01, 12-02, 12-03, 12-04, 12-05) in which CIRM has specifically written that Prop 71 stem cell research is not allowed to apply (evidences if you’d like). If those safety precautions of CIRM to preempt Prop 71 bond prior to ICOC public meetings have failed, Alan Trounson even personally denied the existence of such grants. Alan Trounson’ comment on California stem cell report made self-contradicted presumption that university has better sciences or researchers, which is financial conflict of interest evidence itself. University of California, the biggest beneficiary of Prop 71 so far, has not been where the leadership and advances of stem cell research reside in, nor has shown any support to human embryonic stem cell (hESC) research, as evidenced by that UC officials secretly shut down UC hESC research labs. None of CIRM’s excuses to reject or not to consider human embryonic stem cell research grants of Prop 71 has applied to any ICOC awards. Capricor did not have a single piece of scientific data that their cells can become beating heart muscle as we did for human embryonic stem cells (hESCs), or their science or research is better. UC Davis did not have a single piece of scientific data that their cells can regenerate as we did for hESCs, or their science or research is better. STEM did not have a single piece of scientific data that their cells have the capacity/efficiency to repair as we did for hESCs, or their science or research is better. If it is not financial conflicts, we would like to know what it is that those ICOC board members were all awarded $ 20 Ms, while Prop 71 stem cell research could not even pass CIRM pre-application?

Like Alan Trounson said, there is no evidence for financial conflicts of interest. All ICOC board members have followed the symbolic public step to excuse themselves to vote on their own grants, to the extent that none of those ICOC board members’ awards had enough votes required by the law to approve the funding. But those ICOC board members still have their awards passed to receive Prop 71 bond anyway. Maybe we only need to dig little bit deeper for evidences, if take a look at those stem cell meetings keynoted by Alan Trounson or sponsored by CIRM along the courses, such as ISSCR (International society for stem cell research), world stem cell summit, and stem cell meetings on the mesa. How come almost all the invited speakers have preferably been selected by CIRM to receive $Ms of CIRM awards, even though their talks and research have nothing to do with Prop 71 stem cell research? How come none of these so-called stem cell meetings sponsored by CIRM would welcome hESC research, as evident by no hESC research of Prop 71 was ever invited to present. In our experience, hESC research is not only unwelcomed by stem cell meetings sponsored by CIRM, it is in fact, shockingly, on their blacklist. We all know that ISSCR has lobbied a big number of CIRM funding to sponsor their meetings, did ISSCR use those CIRM money to put human embryonic stem cell research on their blacklist? I might be naïve to feel shocked when my poster of hESC research (titled “deriving cardiac elements from pluripotent hESCs for heart reconstitution”) was declined to be presented by ISSCR program chair at 2010 ISSCR meeting in San Francisco when it was perfectly welcomed to be presented in other non-stem cell meetings (see ISSCR poster communications below at SDRMI wordpress). Why? After I emailed to question the fairness of the meeting committee in selecting scientific presentation and restate the importance of our hESC research to the stem cell field, ISSCR responded that my abstract of only 300 words was actually scored by ISSCR committee and it was found that 90% of the abstract restated a problem that everyone recognizes [referred to our openly concerns for the induced pluripotent stem cells (iPS cells)], and no results in the abstract [in fact, 25% of the abstract was devoted to state the results, though no one would put data/results, reflected in the title, in the abstract] (see ISSCR poster communications below at SDRMI wordpress). I was left without any words because I have never heard such thing that meeting committee would score the abstracts, and apparently, CIRM had also picked up ISSCR such financial conflict of interest tradition to score word and space limited pre-applications. I had emailed and called then ISSCR president Irving Weissman of Stanford University, incoming president Fred Gage of Salk Institute, treasure Sean Morrison, no one responded. We all heard talks of Larry Goldstein and some other members of ISSCR founder’s cycle on the stage that had virtually no stem cell research results to present, but they have never been declined by those meetings sponsored by CIRM. Today, restating those shocking financial conflict of interest results that Irving Weissman gave his personal favorite Shinya Yamanaka preferential keynote for iPS cells at the ISSCR meeting, Deepak Srivastava (the ISSCR program chair) of UCSF would receive big funding for iPS cells and heart reprogramming soon from NIH, and ironically, NIH would soon got sued; Irving Weissman gave his wife preferential on stage presentation for their own company, and Stem cell Inc’s adult stem cells of no Prop 71 scientific merits would get special treatment for $40 M from CIRM, let alone equal amount or more that Irving Weissman has already taken for himself from CIRM with virtually no results, I should feel quite lucky that they had not come after me to shut down my hESC research lab at that time.

Legal Assaults on Human Embryonic Stem Cell Research: A Sanford-Burnham Lab Technician’s Blackmail

Everyone knows that those opponents of human embryonic stem cell (hESC) research love to go to court for ridiculous reasons, such as contestable job threats to the superior legal status of a couple of adult stem cell researchers. So much so even a lab technician in Sanford-Burnham knew how to abuse the law. As we announced publication of Regenerative Medicine Startup’s important hESC research advances and breakthroughs as the only available cures/treatments urgent for patients suffering from heart attack and heart disease [see http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3419496/ released by NIH public access and Press Release: Mending the Borken Heart — Towards Clinical Application of Human Embryonic Stem Cell Therapy Derivatives], those opponents of hESC research know their legal status to steal public fund from stem cell research is numbered, even reckon on making baseless false complaints and legal threats to blackmail editors as their last resort to prevent the release of our important hESC research to the scientific community. We are shocked to learn that Andrew Crain, Evan Snyder’s lab technician at Sanford Burnham, made repeat unauthorized offenders behind our back to retract Regenerative Medicine Startup’s original hESC research that he has no knowledge no contribution by false complaints, as if it is being stolen from him; as if Sanford Consortium for Regenerative Medicine, built with California public funds for stem cell research (Prop 71) and required by the law to open to all stem cell researchers in San Diego scientific community, only belongs to Andrew’s boss Evan Snyder; as if Andrew Crain, Evan Snyder, Vincent Chen, Jean Loring generated any stem cell research data. In Andrew Crain’s exact reckless bully words --- “--- if not retracted immediately ---, legal action will be strongly pursued by our insitute and the NIH will be notified ---”. To see if Andrew Crain’s complaint has any truth in it or it has violated every canon of scientific conduct, maybe we should ask Andrew Crain, or any other plagiarizers Andrew Crain would like to name, if he can provide a single piece of data in this paper to show it was done by him, or write a interpretation for our data showed in this paper, where the data was from, what was for, how the experiment was designed, what was the question we want to answer, and how did we answer it. Why Andrew Crain or Evan Snyder, or any other repeat offenders Andrew Crain would like to name, dared not to send their complaints to me? May be he is totally aware that he is making false complaints and that making false claims or lying in front of judge is serious scientific misconduct or felony if he goes to court. Andrew Crain, Evan Snyder, Vincent Chen, and Jean Loring and their institutes care nothing about stem cell research, maybe their financial conflicts of interest are the real reason that Andrew Crain made the repeat false complaints to prevent publication and release of important hESC research of Regenerative Medicine Startup. If simply see how Evan Snyder/Vincent Chen/Yang Xu plagiarized Dr. Parsons’ original research data for California Institute for Regenerative Medicine (CIRM) grants without any collaboration agreement or permission of Dr. Parsons, violated every canon of scientific conduct, but they never retract public stem cell research fund they have stolen immediately, WHY? If simply see how Evan Snyder, an adult stem cell researcher claimed himself to be the west coast correspondent of NIH and founder of Southern California Stem Cell Consortium, and Sanford Burnham executives lobbied CIRM to steal hundreds of millions of Prop 71 from hESC research through closely-connected Southern California consortium club members that include Larry Goldstein, Yang Xu, Inder Verma, Jean Loring, Vincent Chen, Catriona Jamieson, Leanne Jones, Kristin Baldwin, Bing Ren, and Clive Svendsen (more can be found listed on Stem Cell Meetings on the Mesa), violated every canon of scientific conduct, but they never retract public stem cell research fund they have stolen immediately, WHY? If simply see how Evan Snyder’s Duke, Florida, Harvard connections would give Robert Wechsler-Reya of Sanford-Burnham and those who have never done any stem cell research $ Ms of CIRM funding as ring leader awards, violated every canon of scientific conduct, but they never retract public stem cell research fund they have stolen immediately, WHY? If simply see how Evan Snyder and Sanford-Burnham executives and their consortium members lobbied CIRM to set up many exclusionary unlawful eligibility criteria and issue conflicts of interest RFAs to make sure public stem cell research to come their way without having to generate any scientific data, violated every canon of scientific conduct and Prop 71, but they never retract public stem cell research fund they have stolen immediately, WHY? If simply see why CIRM would set up many exclusionary unlawful eligibility criteria for their RFAs, such as leadership awards have to be out of state, cell line derivation awards have to be iPS adult cells, faculty awards have to be MD, etc., and who have been the biggest financial beneficiary of such conflicts, violated very canon of Prop 71 and Federal and State Laws. If simply see why Sanford-Burnham/UC connect and their associated Crops (e.g., Viacyte, International Stem Cell Corp, Capricor, ACT, Cedars Sinai, Pfizer, Millipore, Johnson & Johnson) would have seminars, meetings, public propaganda events closely corresponding to CIRM RFAs and awards. If simply see why CIRM would give Vincent Chen $Ms for iPS cell conflict of no Prop 71 scientific merit, while block hESC research of Prop 71 scientific merits, and who would be the financial beneficiary of such conflicts, violated every canon of scientific conduct and Prop 71, but Vincent Chen never retract public stem cell research fund he has stolen immediately, WHY? Voice of Regenerative Medicine (VORM) condemn such reckless bullies who have no knowledge no contribution of scientific research data but scientific misconduct and financial conflicts to falsify statements to prevent publication of hESC research breakthroughs and advances release to the scientific community.

Greed & Debt --- CIRM Fraud Manufacturing Machine

Maybe we do not know California Institute for Regenerative Medicine (CIRM) board conflict of interest connections, maybe we do not know CIRM scientific review board conflict of interest connects, maybe we do not know Irv Weissman/Alan Trounson/ISSCR university corruption network connections, maybe we do not know Duane Roth/Connect Washington lobby connections, maybe we do not those glorious cheaters FDA/NIH/Corp alliance connections, but we do know for sure those CIRM awards bearing CA $ 1 billion in debt by greed & debt STEM & Capricor & UC are not stem cell research of Prop 71 not hope for patients CA voted. No scientific evidence $STEM/Weissman adult neural stem cells can work on chronic patients & Alzheimer, no scientific evidence Capricor tissue adult cells can become heart cells regenerate heart muscle as falsified by CIRM grant review statements. CIRM block CA prop71 stem cell research urgent for patients from lawful consideration, but give their board member $STEM/Capricor/UC frauds & scams special treatment against the law. CIRM not only has falsified the $STEM/Capricor/UC grant review statements against scientific evidences, but has not followed 2/3 quorum of Prop71 required for all non pluripotent stem/progenitor cell research. $STEM has cheated investors for > 10 years, now corrupt CA state agency cheat CA Prop71 public stem cell research fund bankrupt CA. STEM/Irv Weissman/SU have 7 conflict-of-interest CIRM bureaucrat connections do not mean STEM openly corrupt CA state agency is ok, CIRM board anti-law anti-ethics anti-moral is ok. Dumb wall street investors throw your money into stock frauds we care none, but STEM/Weissman/Capricor/UC corrupt CA state agency corrupt moral/ethics to cheat CA Prop 71 stem cell research fund we care VORM care patients care law care. Stop greed & debt STEM & Corp corrupt CA state agency cheat Prop 71.