305 results on '"Evan Y. Snyder"'
Search Results
2. Discovery of suppressors of CRMP2 phosphorylation reveals compounds that mimic the behavioral effects of lithium on amphetamine-induced hyperlocomotion
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Wen-Ning Zhao, Brian T. D. Tobe, Namrata D. Udeshi, Lucius L. Xuan, Cameron D. Pernia, Daniel P. Zolg, Amanda J. Roberts, Deepak Mani, Sarah R. Blumenthal, Iren Kurtser, Debasis Patnaik, Irina Gaisina, Joshua Bishop, Steven D. Sheridan, Jasmin Lalonde, Steven A. Carr, Evan Y. Snyder, and Stephen J. Haggarty
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Abstract The effective treatment of bipolar disorder (BD) represents a significant unmet medical need. Although lithium remains a mainstay of treatment for BD, limited knowledge regarding how it modulates affective behavior has proven an obstacle to discovering more effective mood stabilizers with fewer adverse side effects. One potential mechanism of action of lithium is through inhibition of the serine/threonine protein kinase GSK3β, however, relevant substrates whose change in phosphorylation may mediate downstream changes in neuroplasticity remain poorly understood. Here, we used human induced pluripotent stem cell (hiPSC)-derived neuronal cells and stable isotope labeling by amino acids in cell culture (SILAC) along with quantitative mass spectrometry to identify global changes in the phosphoproteome upon inhibition of GSK3α/β with the highly selective, ATP-competitive inhibitor CHIR-99021. Comparison of phosphorylation changes to those induced by therapeutically relevant doses of lithium treatment led to the identification of collapsin response mediator protein 2 (CRMP2) as being highly sensitive to both treatments as well as an extended panel of structurally distinct GSK3α/β inhibitors. On this basis, a high-content image-based assay in hiPSC-derived neurons was developed to screen diverse compounds, including FDA-approved drugs, for their ability to mimic lithium’s suppression of CRMP2 phosphorylation without directly inhibiting GSK3β kinase activity. Systemic administration of a subset of these CRMP2-phosphorylation suppressors were found to mimic lithium’s attenuation of amphetamine-induced hyperlocomotion in mice. Taken together, these studies not only provide insights into the neural substrates regulated by lithium, but also provide novel human neuronal assays for supporting the development of mechanism-based therapeutics for BD and related neuropsychiatric disorders.
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- 2020
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3. The Developmental & Molecular Requirements for Ensuring that Human Pluripotent Stem Cell-Derived Hair Follicle Bulge Stem Cells Have Acquired Competence for Hair Follicle Generation Following Transplantation
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Michel R. Ibrahim, Walid Medhat, Hasan El-Fakahany, Hamza Abdel-Raouf, and Evan Y. Snyder
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Medicine - Abstract
When using human induced pluripotent stem cells (hiPSCs) to achieve hair follicle (HF) replacement, we found it best to emulate the earliest fundamental developmental processes of gastrulation, ectodermal lineage commitment, and dermogenesis. Viewing hiPSCs as a model of the epiblast, we exploited insights from mapping the dynamic up- and down-regulation of the developmental molecules that determine HF lineage in order to ascertain the precise differentiation stage and molecular requirements for grafting HF-generating progenitors. To yield an integrin-dependent lineage like the HF in vivo, we show that hiPSC derivatives should co-express, just prior to transplantation, the following combination of markers: integrins α6 and β1 and the glycoprotein CD200 on their surface; and, intracellularly, the epithelial marker keratin 18 and the hair follicle bulge stem cell (HFBSC)-defining molecules transcription factor P63 and the keratins 15 and 19. If the degree of trichogenic responsiveness indicated by the presence of these molecules is not achieved (they peak on Days 11-18 of the protocol), HF generation is not possible. Conversely, if differentiation of the cells is allowed to proceed beyond the transient intermediate progenitor state represented by the HFBSC, and instead cascades to their becoming keratin 14 + keratin 5 + CD200 – keratinocytes (Day 25), HF generation is equally impossible. We make the developmental case for transplanting at Day 16-18 of differentiation—the point at which the hiPSCs have lost pluripotency, have attained optimal expression of HFBSC markers, have not yet experienced downregulation of key integrins and surface glycoproteins, have not yet started expressing keratinocyte-associated molecules, and have sufficient proliferative capacity to allow a well-populated graft. This panel of markers may be used for isolating (by cytometry) HF-generating derivatives away from cell types unsuited for this therapy as well as for identifying trichogenic drugs.
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- 2021
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4. Nna1 gene deficiency triggers Purkinje neuron death by tubulin hyperglutamylation and ER dysfunction
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Jianxue Li, Evan Y. Snyder, Fenny H.F. Tang, Renata Pasqualini, Wadih Arap, and Richard L. Sidman
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Neuroscience ,Medicine - Abstract
Posttranslational glutamylation/deglutamylation balance in tubulins influences dendritic maturation and neuronal survival of cerebellar Purkinje neurons (PNs). PNs and some additional neuronal types degenerate in several spontaneous, independently occurring Purkinje cell degeneration (pcd) mice featuring mutant neuronal nuclear protein induced by axotomy (Nna1), a deglutamylase gene. This defective deglutamylase allows glutamylases to form hyperglutamylated tubulins. In pcd, all PNs die during postnatal “adolescence.” Neurons in some additional brain regions also die, mostly later than PNs. We show in laser capture microdissected single PNs, in cerebellar granule cell neuronal clusters, and in dissected hippocampus and substantia nigra that deglutamase mRNA and protein were virtually absent before pcd PNs degenerated, whereas glutaminase mRNA and protein remained normal. Hyperglutamylated microtubules and dimeric tubulins accumulated in pcd PNs and were involved in pcd PN death by glutamylase/deglutamylase imbalance. Importantly, treatment with a microtubule depolymerizer corrected the glutamylation/deglutamylation ratio, increasing PN survival. Further, before onset of neuronal death, pcd PNs displayed prominent basal polylisosomal masses rich in ER. We propose a “seesaw” metamorphic model summarizing mutant Nna1-induced tubulin hyperglutamylation, the pcd’s PN phenotype, and report that the neuronal disorder involved ER stress, unfolded protein response, and protein synthesis inhibition preceding PN death by apoptosis/necroptosis.
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- 2020
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5. Driving Neuronal Differentiation through Reversal of an ERK1/2-miR-124-SOX9 Axis Abrogates Glioblastoma Aggressiveness
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Hanna Sabelström, Rebecca Petri, Ksenya Shchors, Rahul Jandial, Christin Schmidt, Rohit Sacheva, Selma Masic, Edith Yuan, Trenten Fenster, Michael Martinez, Supna Saxena, Theodore P. Nicolaides, Shirin Ilkhanizadeh, Mitchel S. Berger, Evan Y. Snyder, William A. Weiss, Johan Jakobsson, and Anders I. Persson
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Biology (General) ,QH301-705.5 - Abstract
Summary: Identifying cellular programs that drive cancers to be stem-like and treatment resistant is critical to improving outcomes in patients. Here, we demonstrate that constitutive extracellular signal-regulated kinase 1/2 (ERK1/2) activation sustains a stem-like state in glioblastoma (GBM), the most common primary malignant brain tumor. Pharmacological inhibition of ERK1/2 activation restores neurogenesis during murine astrocytoma formation, inducing neuronal differentiation in tumorspheres. Constitutive ERK1/2 activation globally regulates miRNA expression in murine and human GBMs, while neuronal differentiation of GBM tumorspheres following the inhibition of ERK1/2 activation requires the functional expression of miR-124 and the depletion of its target gene SOX9. Overexpression of miR124 depletes SOX9 in vivo and promotes a stem-like-to-neuronal transition, with reduced tumorigenicity and increased radiation sensitivity. Providing a rationale for reports demonstrating miR-124-induced abrogation of GBM aggressiveness, we conclude that reversal of an ERK1/2-miR-124-SOX9 axis induces a neuronal phenotype and that enforcing neuronal differentiation represents a therapeutic strategy to improve outcomes in GBM. : Sabelström et al. show that the loss of neurogenesis is reversible during neural stem cell-derived glioma formation. Pharmacological inhibition of ERK1/2 globally regulates miRNAs and induces neuronal differentiation, a process that is dependent on the modulation of an miR-124-SOX9 axis in glioblastoma (GBM) cells. The overexpression of miR-124 induces neuronal differentiation that abrogates GBM aggressiveness. Keywords: brain, cancer, differentiation, glioma, glioblastoma, microRNA, neural stem cell, neurogenesis, neuron, tumor
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- 2019
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6. Quantitative Analysis of Human Pluripotency and Neural Specification by In-Depth (Phospho)Proteomic Profiling
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Ilyas Singec, Andrew M. Crain, Junjie Hou, Brian T.D. Tobe, Maria Talantova, Alicia A. Winquist, Kutbuddin S. Doctor, Jennifer Choy, Xiayu Huang, Esther La Monaca, David M. Horn, Dieter A. Wolf, Stuart A. Lipton, Gustavo J. Gutierrez, Laurence M. Brill, and Evan Y. Snyder
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Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
Controlled differentiation of human embryonic stem cells (hESCs) can be utilized for precise analysis of cell type identities during early development. We established a highly efficient neural induction strategy and an improved analytical platform, and determined proteomic and phosphoproteomic profiles of hESCs and their specified multipotent neural stem cell derivatives (hNSCs). This quantitative dataset (nearly 13,000 proteins and 60,000 phosphorylation sites) provides unique molecular insights into pluripotency and neural lineage entry. Systems-level comparative analysis of proteins (e.g., transcription factors, epigenetic regulators, kinase families), phosphorylation sites, and numerous biological pathways allowed the identification of distinct signatures in pluripotent and multipotent cells. Furthermore, as predicted by the dataset, we functionally validated an autocrine/paracrine mechanism by demonstrating that the secreted protein midkine is a regulator of neural specification. This resource is freely available to the scientific community, including a searchable website, PluriProt.
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- 2016
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7. Novel Bivalent and D-Peptide Ligands of CXCR4 Mobilize Hematopoietic Progenitor Cells to the Blood in C3H/HeJ Mice
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Yujia Mao, Qian Meng, Panpan Song, Siyu Zhu, Yan Xu, Evan Y. Snyder, Jing An, and Ziwei Huang
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Medicine - Abstract
The interaction of SDF-1α (also known as CXCL12) with the CXCR4 receptor plays a critical role in the retention of hematopoietic stem cells (HSCs) in bone marrow. The viral macrophage inflammatory protein-II (vMIP-II), a human herpesvirus-8 (HHV-8)-encoded viral chemokine, can bind the CXCR4 receptor and inhibit endogenous ligand-induced calcium responses and cell migration. Previously, we used the bivalent ligand approach to link synthetically two unnatural D-amino acid peptides derived from the N-terminus of vMIP-II (DV1 and DV3, respectively) to generate a dimeric peptide, DV1-K-(DV3) (also named HC4319), which shows very high affinity for CXCR4. Here, we studied the biological effects of this dimeric peptide, HC4319, and its monomeric counterpart, DV1, on SDF-1α-induced signaling in CXCR4- or CXCR7-transfected Chinese hamster ovary cells and mobilization of hematopoietic progenitor cells (HPCs) in C3H/HeJ mice using an HPC assay. HC4319 and DV1 inhibited significantly the phosphorylation of Akt and Erk, known to be downstream signaling events of CXCR4. This in vivo study in C3H/HeJ mice showed that HC4319 and DV-1 strongly induced rapid mobilization of granulocyte–macrophage colony-forming units (CFUs), erythrocyte burst-forming units, and granulocyte–erythrocyte–monocyte–megakaryocyte CFUs from the bone marrow to the blood. These results provide the first reported experimental evidence that bivalent and D-amino acid peptides derived from the N-terminus of vMIP-II are potent mobilizers of HPCs in C3H/HeJ mice and support the further development of such agents for clinical application.
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- 2018
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8. hESC Differentiation toward an Autonomic Neuronal Cell Fate Depends on Distinct Cues from the Co-Patterning Vasculature
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Lisette M. Acevedo, Jeffrey N. Lindquist, Breda M. Walsh, Peik Sia, Flavio Cimadamore, Connie Chen, Martin Denzel, Cameron D. Pernia, Barbara Ranscht, Alexey Terskikh, Evan Y. Snyder, and David A. Cheresh
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Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
To gain insight into the cellular and molecular cues that promote neurovascular co-patterning at the earliest stages of human embryogenesis, we developed a human embryonic stem cell model to mimic the developing epiblast. Contact of ectoderm-derived neural cells with mesoderm-derived vasculature is initiated via the neural crest (NC), not the neural tube (NT). Neurovascular co-patterning then ensues with specification of NC toward an autonomic fate requiring vascular endothelial cell (EC)-secreted nitric oxide (NO) and direct contact with vascular smooth muscle cells (VSMCs) via T-cadherin-mediated homotypic interactions. Once a neurovascular template has been established, NT-derived central neurons then align themselves with the vasculature. Our findings reveal that, in early human development, the autonomic nervous system forms in response to distinct molecular cues from VSMCs and ECs, providing a model for how other developing lineages might coordinate their co-patterning.
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- 2015
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9. Neural Stem Cells Derived from Human Parthenogenetic Stem Cells Engraft and Promote Recovery in a Nonhuman Primate Model of Parkinson's Disease
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Rodolfo Gonzalez, Ibon Garitaonandia, Maxim Poustovoitov, Tatiana Abramihina, Caleb McEntire, Ben Culp, Jordan Attwood, Alexander Noskov, Trudy Christiansen-Weber, Marwa Khater, Sergio Mora-Castilla, Cuong To, Andrew Crain, Glenn Sherman, Andrey Semechkin, Louise C. Laurent, John D. Elsworth, John Sladek, Evan Y. Snyder, D. Eugene Redmond, and Russell A. Kern
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Medicine - Abstract
Cell therapy has attracted considerable interest as a promising therapeutic alternative for patients with Parkinson's disease (PD). Clinical studies have shown that grafted fetal neural tissue can achieve considerable biochemical and clinical improvements in PD. However, the source of fetal tissue grafts is limited and ethically controversial. Human parthenogenetic stem cells offer a good alternative because they are derived from unfertilized oocytes without destroying potentially viable human embryos and can be used to generate an unlimited supply of neural cells for transplantation. We have previously reported that human parthenogenetic stem cell-derived neural stem cells (hpNSCs) successfully engraft, survive long term, and increase brain dopamine (DA) levels in rodent and nonhuman primate models of PD. Here we report the results of a 12-month transplantation study of hpNSCs in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned African green monkeys with moderate to severe clinical parkinsonian symptoms. The hpNSCs manufactured under current good manufacturing practice (cGMP) conditions were injected bilaterally into the striatum and substantia nigra of immunosuppressed monkeys. Transplantation of hpNSCs was safe and well tolerated by the animals with no dyskinesia, tumors, ectopic tissue formation, or other test article-related serious adverse events. We observed that hpNSCs promoted behavioral recovery; increased striatal DA concentration, fiber innervation, and number of dopaminergic neurons; and induced the expression of genes and pathways downregulated in PD compared to vehicle control animals. These results provide further evidence for the clinical translation of hpNSCs and support the approval of the world's first pluripotent stem cell-based phase I/IIa study for the treatment of PD (Clinical Trial Identifier NCT02452723).
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- 2016
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10. Proof of Concept Studies Exploring the Safety and Functional Activity of Human Parthenogenetic-Derived Neural Stem Cells for the Treatment of Parkinson's Disease
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Rodolfo Gonzalez, Ibon Garitaonandia, Andrew Crain, Maxim Poustovoitov, Tatiana Abramihina, Alexander Noskov, Chuan Jiang, Robert Morey, Louise C. Laurent, John D. Elsworth, Evan Y. Snyder, D. Eugene Redmond, and Ruslan Semechkin
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Medicine - Abstract
Recent studies indicate that human pluripotent stem cell (PSC)-based therapies hold great promise in Parkinson's disease (PD). Clinical studies have shown that grafted fetal neural tissue can achieve considerable biochemical and clinical improvements in PD. However, the source of fetal tissue grafts is limited and ethically controversial. Human parthenogenetic stem cells offer a good alternative because they are derived from unfertilized oocytes without destroying viable human embryos and can be used to generate an unlimited supply of neural stem cells for transplantation. Here we evaluate for the first time the safety and engraftment of human parthenogenetic stem cell-derived neural stem cells (hpNSCs) in two animal models: 6-hydroxydopamine (6-OHDA)-lesioned rodents and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated nonhuman primates (NHPs). In both rodents and nonhuman primates, we observed successful engraftment and higher dopamine levels in hpNSC-transplanted animals compared to vehicle control animals, without any adverse events. These results indicate that hpNSCs are safe, well tolerated, and could potentially be a source for cell-based therapies in PD.
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- 2015
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11. The Use of Nonneuronal Cells for Gene Delivery
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Evan Y. Snyder and Marie-Claude Senut
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Published
- 1997
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12. DMSO-Free Programmed Cryopreservation of Fully Dissociated and Adherent Human Induced Pluripotent Stem Cells
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Igor I. Katkov, Natalia G. Kan, Flavio Cimadamore, Brandon Nelson, Evan Y. Snyder, and Alexey V. Terskikh
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Internal medicine ,RC31-1245 - Abstract
Three modes for cryopreservation (CP) of human iPSC cells have been compared: STD: standard CP of small clumps with 10% of CPA in cryovials, ACC: dissociation of the cells with Accutase and freezing in cryovials, and PLT: programmed freezing of adherent cells in plastic multiwell dishes in a programmable freezer using one- and multistep cooling protocols. Four CPAs were tesetd: dimethyl sulfoxide (DMSO), ethylene glycol (EG), propylene glycol (PG), and glycerol (GLY). The cells in ACC and PLT were frozen and recovered after thawing in the presence of a ROCK inhibitor Y-27632 (RI). EG was less toxic w/o CP cryopreservation than DMSO and allowed much better maintenance of pluripotency after CP than PG or GLY. The cells were cryopreserved very efficiently as adherent cultures (+RI) in plates (5-6-fold higher than STD) using EG and a 6-step freezing protocol. Recovery under these conditions is comparable or even higher than ACC+RI. Conclusions. Maintenance of cell-substratum adherence is a favorable environment that mitigates freezing and thawing stresses (ComfortFreeze® concept developed by CELLTRONIX). CP of cells directly in plates in ready-to-go after thawing format for HT/HC screening can be beneficial in many SC-related scientific and commercial applications such as drug discovery and toxicity tests.
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- 2011
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13. Neural Stem Cells Implanted into MPTP-Treated Monkeys Increase the Size of Endogenous Tyrosine Hydroxylase-Positive Cells Found in the Striatum: A Return to Control Measures
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Kimberly B. Bjugstad, D. Eugene Redmond, Yang D. Teng, J. D. Elsworth, R. H. Roth, B. C. Blanchard, Evan Y. Snyder, and John R. Sladek
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Medicine - Abstract
Neural stem cells (NSC) have been shown to migrate towards damaged areas, produce trophic factors, and replace lost cells in ways that might be therapeutic for Parkinson's disease (PD). However, there is very little information on the effects of NSC on endogenous cell populations. In the current study, effects of implanted human NSC (hNSC) on endogenous tyrosine hydroxylase-positive cells (TH+ cells) after treatment with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were explored in nonhuman primates. After MPTP damage and in PD, the primate brain is characterized by decreased numbers of dopamine neurons in the substantia nigra (SN) and an increase in neurons expressing TH in the caudate nucleus. To determine how implanted NSC might affect these cell populations, 11 St. Kitts African green monkeys were treated with the selective dopaminergic neurotoxin, MPTP. Human NSC were implanted into the left and right caudate nucleus and the right SN of eight of the MPTP-treated monkeys. At either 4 or 7 months after NSC implants, the brains were removed and the size and number of TH+ cells in the target areas were assessed. The results were compared to data obtained from normal untreated control monkeys and to the three unimplanted MPTP-treated monkeys. The majority of hNSC were found bilaterally along the nigrostriatal pathway and in the substantia nigra, while relatively few were found in the caudate. In the presence of NSC, the number and size of caudate TH+ cells returned to non-MPTP-treated control levels. MPTP-induced and hNSC-induced changes in the putamen were less apparent. We conclude that after MPTP treatment in the primate, hNSC prevent the MPTP-induced upregulation of TH+ cells in the caudate and putamen, indicating that hNSC may be beneficial to maintaining a normal striatal environment.
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- 2005
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14. The lung employs an intrinsic surfactant-mediated inflammatory response for viral defense
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Sandra L. Leibel, Rachael N. McVicar, Rabi Murad, Elizabeth M. Kwong, Alex E. Clark, Asuka Alvarado, Bethany A. Grimmig, Ruslan Nuryyev, Randee E. Young, Jamie Casey Lee, Weiqi Peng, Yanfang Peipei Zhu, Eric Griffis, Cameron J. Nowell, Kang Liu, Brian James, Suzie Alarcon, Atul Malhotra, Linden J. Gearing, Paul J. Hertzog, Cheska Marie Galapate, Koen M.O. Galenkamp, Cosimo Commisso, Davey M. Smith, Xin Sun, Aaron F. Carlin, Ben A. Croker, and Evan Y. Snyder
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Article - Abstract
Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) causes an acute respiratory distress syndrome (ARDS) that resembles surfactant deficient RDS. Using a novel multi-cell type, human induced pluripotent stem cell (hiPSC)-derived lung organoid (LO) system, validated against primary lung cells, we found that inflammatory cytokine/chemokine production and interferon (IFN) responses are dynamically regulated autonomously within the lung following SARS-CoV-2 infection, an intrinsic defense mechanism mediated by surfactant proteins (SP). Single cell RNA sequencing revealed broad infectability of most lung cell types through canonical (ACE2) and non-canonical (endocytotic) viral entry routes. SARS-CoV-2 triggers rapid apoptosis, impairing viral dissemination. In the absence of surfactant protein B (SP-B), resistance to infection was impaired and cytokine/chemokine production and IFN responses were modulated. Exogenous surfactant, recombinant SP-B, or genomic correction of the SP-B deletion restored resistance to SARS-CoV-2 and improved viability.
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- 2023
15. A newly-recognized population of residual neural crest cells in the adult leptomeninges is re-activated for vascular repair
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Yoshihiko Nakamura, Takafumi Nakano, Ji Hyun Park, Masayoshi Tanaka, Wenlu Li, Elga Esposito, Bum Ju Ahn, Violeta Durán-Laforet, Rakhi Desai, Ikbal Sencan, Sava Sakadžić, Eng H. Lo, Evan Y. Snyder, Marcin Tabaka, and Kazuhide Hayakawa
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SummaryThe neural crest (NC) is a transient structure in vertebrate embryogenesis comprising highly migratory multipotent stem cells that give rise to a diverse array of cell types in organs throughout the body, including initiating neurovascular patterning. It is assumed that neural crest stem cells (NCSCs) disappear after development. Unexpectedly, using single-nucleus RNA-sequencing, we discovered residual quiescent NCSCs in the adult mouse meninges which are activated by injury and contribute to the brain’s homeostatic response. RNA velocity, pathway, and transcription factor analyses in a murine stroke model (combined with in vivo imaging) show that these adult NCSCs migrate towards the perivascular spaces of the infarct and undergo a perivascular stromal cell transition that is regulated by Ptp1b, Ghr, and Stat3. Loss- and gain-of-function experiments show that these “vestigial” NCSCs are required for restoring vascular endothelial barrier function via β-catenin and Stat3 signaling. These findings suggest that, in the adult, an unexpected reservoir of cells -- once pivotal to embryogenesis and vascular morphogenesis -- are re-invoked for neurovascular repair.
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- 2022
16. Meningeal Multipotent Cells: A Hidden Target for CNS Repair?
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Kazuhide Hayakawa, Eng H. Lo, and Evan Y. Snyder
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Central Nervous System ,Male ,0301 basic medicine ,Central nervous system ,Biology ,Article ,Brain Ischemia ,Rats, Sprague-Dawley ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Meninges ,0302 clinical medicine ,Cerebrospinal fluid ,Neural Stem Cells ,Central Nervous System Diseases ,medicine ,Animals ,Homeostasis ,Humans ,Regeneration ,Adapalene ,Perivascular space ,Multipotent Stem Cells ,Mesenchymal stem cell ,Neural crest ,Rats ,Adult Stem Cells ,030104 developmental biology ,medicine.anatomical_structure ,Neurology ,Neural Crest ,Multipotent Stem Cell ,Molecular Medicine ,Stem cell ,Glymphatic System ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Traditionally, the primary role of the meninges is thought to be structural, i.e., to act as a surrounding membrane that contains and cushions the brain with cerebrospinal fluid. During development, the meninges is formed by both mesenchymal and neural crest cells. There is now emerging evidence that subsets of undifferentiated stem cells might persist in the adult meninges. In this mini-review, we survey representative studies of brain-meningeal interactions and discuss the hypothesis that the meninges are not just protective membranes, but instead contain multiplex stem cell subsets that may contribute to central nervous system (CNS) homeostasis. Further investigations into meningeal multipotent cells may reveal a “hidden” target for promoting neurovascular remodeling and repair after CNS injury and disease.
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- 2021
17. Strategies and Progress in CXCR4-Targeted Anti-Human Immunodeficiency Virus (HIV) Therapeutic Development
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Robert T. Schooley, Evan Y. Snyder, and Lina S M Huang
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0301 basic medicine ,Microbiology (medical) ,Receptors, CXCR4 ,Proteases ,Receptors, CCR5 ,Cell ,HIV Infections ,CXCR4 ,03 medical and health sciences ,Chemokine receptor ,0302 clinical medicine ,Acquired immunodeficiency syndrome (AIDS) ,Viral entry ,medicine ,Humans ,Receptor ,Acquired Immunodeficiency Syndrome ,business.industry ,virus diseases ,medicine.disease ,Virology ,Reverse transcriptase ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,HIV-1 ,business - Abstract
The acquired immunodeficiency syndrome (AIDS), caused by the human immunodeficiency virus (HIV), has been a global public health challenge for several decades. The majority of HIV infection is caused by the human immunodeficiency virus type 1 (HIV-1), which enters and infects a host cell via the cell surface proteins of CD4 as the primary receptor, and chemokine receptors CXCR4 or CCR5 as the coreceptor–then undergoing replication using the cell’s intracellular machinery. Whereas many drugs targeting CCR5-mediated entry or HIV-1 replication via reverse transcriptase or proteases have long been used clinically, agents targeting CXCR4 are yet to be advanced to clinical application. Here in this review we highlight some of the strategies for and progress made in the discovery of novel small molecules, peptides, and larger molecules that target CXCR4, and their future prospects for translation into the clinic as a new class of anti-HIV therapeutics.
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- 2021
18. Valproate reverses mania-like behaviors in mice via preferential targeting of HDAC2
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Evan Y. Snyder, Angela R. Ozburn, Colleen A. McClung, Mariah A. Hildebrand, George C. Tseng, Jeffrey Oliver-Smith, Matthew B Jarpe, Alicia M. Winquist, Hui Zhang, Wei Zong, Puja K. Parekh, Darius Becker-Krail, Zhiguang Huo, Ethan Fitzgerald, Rachel N. Arey, Xiyu Zhu, Lauren M. DePoy, Andrew Crain, Kyle D. Ketchesin, Brian T. D. Tobe, Xiangning Xue, and Ryan W. Logan
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0301 basic medicine ,Induced Pluripotent Stem Cells ,Histone Deacetylase 2 ,Pharmacology ,Article ,Treatment of bipolar disorder ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Mice ,0302 clinical medicine ,RNA interference ,mental disorders ,Medicine ,Animals ,Humans ,Induced pluripotent stem cell ,Molecular Biology ,business.industry ,Histone deacetylase 2 ,Valproic Acid ,HDAC1 ,Ventral tegmental area ,Histone Deacetylase Inhibitors ,Psychiatry and Mental health ,Mania ,030104 developmental biology ,medicine.anatomical_structure ,lipids (amino acids, peptides, and proteins) ,Histone deacetylase ,medicine.symptom ,business ,030217 neurology & neurosurgery - Abstract
Valproate (VPA) has been used in the treatment of bipolar disorder since the 1990s. However, the therapeutic targets of VPA have remained elusive. Here we employ a preclinical model to identify the therapeutic targets of VPA. We find compounds that inhibit histone deacetylase proteins (HDACs) are effective in normalizing manic-like behavior, and that class I HDACs (e.g., HDAC1 and HDAC2) are most important in this response. Using an RNAi approach, we find that HDAC2, but not HDAC1, inhibition in the ventral tegmental area (VTA) is sufficient to normalize behavior. Furthermore, HDAC2 overexpression in the VTA prevents the actions of VPA. We used RNA sequencing in both mice and human induced pluripotent stem cells (iPSCs) derived from bipolar patients to further identify important molecular targets. Together, these studies identify HDAC2 and downstream targets for the development of novel therapeutics for bipolar mania.
- Published
- 2020
19. Induced pluripotency and spontaneous reversal of cellular aging in supercentenarian donor cells
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Paola A. Bignone, Dana Larocca, L.S. Coles, Evan Y. Snyder, Yang Liu, and Ji-Eun Lee
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Adult ,0301 basic medicine ,Induced Pluripotent Stem Cells ,Population ,Biophysics ,Clone (cell biology) ,Biology ,Biochemistry ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Progenitor cell ,Child ,education ,Induced pluripotent stem cell ,Molecular Biology ,Cellular Senescence ,Aged, 80 and over ,Progeria ,education.field_of_study ,Mesenchymal stem cell ,Telomere Homeostasis ,Cell Differentiation ,Mesenchymal Stem Cells ,Cell Biology ,Cellular Reprogramming ,medicine.disease ,Tissue Donors ,Clone Cells ,Cell biology ,Telomere ,030104 developmental biology ,Gene Expression Regulation ,030220 oncology & carcinogenesis ,Transcriptome ,Reprogramming - Abstract
Supercentenarians (≥110-year-old, SC) are a uniquely informative population not only because they surpass centenarians in age, but because they appear to age more slowly with fewer incidences of chronic age-related disease than centenarians. We reprogramed donor B-lymphoblastoid cell lines (LCL) derived from a 114-year-old (SC), a 43-year-old healthy disease-free control (HDC) and an 8-year-old with a rapid aging disease (Hutchinson-Gilford progeria syndrome (HGPS)) and compared SC-iPSC to HDC-iPSC and HGPS-iPSCs. Reprogramming to pluripotency was confirmed by pluripotency marker expression and differentiation to 3 germ-layers. Each iPSC clone differentiated efficiently to mesenchymal progenitor cells (MPC) as determined by surface marker expression and RNAseq analysis. We identified supercentenarian and HGPS associated gene expression patterns in the differentiated MPC lines that were not evident in the parental iPSC lines. Importantly, telomere length resetting occurred in iPSC from all donors albeit at a lower incidence in supercentenarian iPSCs. These data indicate the potential to use reprogramming to reset both developmental state and cellular age in the "oldest of the old." We anticipate that supercentenarian iPSC and their differentiated derivatives will be valuable tools for studying the underlying mechanisms of extreme longevity and disease resistance.
- Published
- 2020
20. Contributors
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K.I. Agladze, Ibrahim Akin, Michail A. Alterman, Abdolreza Ardeshirylajimi, Alessia Bertero, Rahulkumar Bhoi, Natacha Breuls, Anne Bush, Francesca Caloni, Ivan Carcamo-Orive, Zhifen Chen, Teresa Coccini, Massimo Conese, Chad A. Cowan, Nathan James Cunningham, Ibrahim El-Battrawy, Benjamin S. Freedman, Andrée Gauthier-Fisher, Melkamu Getie-Kebtie, Nefele Giarratana, Madelyn A. Gillentine, Christopher Grunseich, Mirabelle S.H. Ho, Miriel S.H. Ho, Deborah A. Hursh, Gentaro Ikeda, Ji Hye Jung, Martin H. Kang, Yong Kyun Kim, Jacqueline Kowitz, Siegfried Lang, Onofrio Laselva, Jennifer Lei, Sandra L. Leibel, Clifford L. Librach, Geoffrey P. Lomax, Maryam Mahmoodinia Maymand, Rachael N. McVicar, Andrew R. Mendelsohn, Anna R. Mendelsohn, Sadegh lotfalah Moradi, Yasuhiro Murakawa, Dan Nir, Connor G. O'Brien, A.D. Podgurskaya, Natalia S. Pripuzova, Maurilio Sampaolesi, M.M. Slotvitsky, Evan Y. Snyder, Shi Su, Colin Sweeney, Bernard Thébaud, V.A. Tsvelaya, Haritha Vallabhaneni, Evgeniya A. Vaskova, Huaxiao Yang, Phillip C. Yang, Laura Yedigaryan, Masahito Yoshihara, and Xiaobo Zhou
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- 2022
21. Using human induced pluripotent stem cells to advance personalized/precision medicine
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Sandra L. Leibel, Rachael N. McVicar, Anne Bush, and Evan Y. Snyder
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- 2022
22. Clinical evidence that a dysregulated master neural network modulator may aid in diagnosing schizophrenia
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Joseph T. Coyle, Richard L. Sidman, Brian T. D. Tobe, Aoi Jitsuki-Takahashi, Hiroko Makihara, Fumio Nakamura, Haruko Nakamura, Yoshio Hirayasu, Keisuke Watanabe, Naoya Yamashita, Glenn T. Konopaske, Evan Y. Snyder, Cameron D Pernia, Yoshio Goshima, Munetaka Nomoto, Reina Aoki, Yusuke Saigusa, Toshihiko Baba, Francine M. Benes, and Mari Saito
- Subjects
0301 basic medicine ,medicine.medical_specialty ,collapsin response mediator protein-2 (CRMP2) ,Nerve Tissue Proteins ,blood test ,03 medical and health sciences ,0302 clinical medicine ,Mediator ,Internal medicine ,medicine ,Biological neural network ,Blood test ,Humans ,Bipolar disorder ,Multidisciplinary ,medicine.diagnostic_test ,business.industry ,cytoskeleton ,Biological Sciences ,medicine.disease ,dendritic morphology ,030104 developmental biology ,Endocrinology ,Gene Expression Regulation ,Schizophrenia ,Biomarker (medicine) ,biomarker ,Intercellular Signaling Peptides and Proteins ,Collapsin response mediator protein family ,medicine.symptom ,Nerve Net ,business ,Mania ,030217 neurology & neurosurgery ,Biomarkers ,Neuroscience ,Genome-Wide Association Study - Abstract
Significance There are no biomarkers for schizophrenia (SCZ), a disorder of dysfunctional neural networks. We demonstrate that a master regulator of cytoskeleton (“CRMP2”) and, hence, neural circuitry, may form the basis for such a biomarker because its activity is uniquely imbalanced in SCZ patients. We show that SCZ patients are characterized by an excess of active CRMP2 not only in their brains (where it is correlated with dendritic abnormalities) but also in their peripheral blood lymphocytes. The abundance of active CRMP2 and insufficiency of opposing inactive p-CRMP2 likely disrupts neuronal function. Because peripheral blood CRMP2 appears to reflect intracerebral processes, it could form the basis of a rapid, minimally invasive, sensitive, and specific clinical diagnostic aid for SCZ in young patients., There are no validated biomarkers for schizophrenia (SCZ), a disorder linked to neural network dysfunction. We demonstrate that collapsin response mediator protein-2 (CRMP2), a master regulator of cytoskeleton and, hence, neural circuitry, may form the basis for a biomarker because its activity is uniquely imbalanced in SCZ patients. CRMP2’s activity depends upon its phosphorylation state. While an equilibrium between inactive (phosphorylated) and active (nonphosphorylated) CRMP2 is present in unaffected individuals, we show that SCZ patients are characterized by excess active CRMP2. We examined CRMP2 levels first in postmortem brains (correlated with neuronal morphometrics) and then, because CRMP2 is expressed in lymphocytes as well, in the peripheral blood of SCZ patients versus age-matched unaffected controls. In the brains and, more starkly, in the lymphocytes of SCZ patients
- Published
- 2021
23. A Tool for Accurate Stoichiometric Composition of Cryopreservative Media for Fetal and Induced Pluripotent Stem Cell-Derived Human Neural Stem Cells
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Evan Y. Snyder and Walter D. Niles
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Cryopreservation ,General Immunology and Microbiology ,Cryoprotectant ,Chemistry ,General Neuroscience ,Cell ,Induced Pluripotent Stem Cells ,Health Informatics ,Vitrification ,General Biochemistry, Genetics and Molecular Biology ,Neural stem cell ,Cell biology ,Transplantation ,Medical Laboratory Technology ,medicine.anatomical_structure ,Cryoprotective Agents ,Neural Stem Cells ,medicine ,Humans ,Viability assay ,General Pharmacology, Toxicology and Pharmaceutics ,Induced pluripotent stem cell - Abstract
Fetal human neural stem cells (fhNSC) are of considerable interest as potential regenerative therapies for neuronal or glial degeneration or destruction resulting from genetic abnormalities, disease, or injury. Realization of this potential requires securing a supply of cells sufficient to meet the needs of transplantation, which are often tens to hundreds of millions of cells per dose. This challenge necessitates the establishment of safe and efficient cell banking protocols. Cryopreservation, involving the slow freezing or vitrification of cells, enables storage of fhNSC for prolonged periods, while maintaining their viability and multipotency required for clinical use. To optimize cryopreservation of fhNSC, attention has become focused on the composition of the medium used to effect cryopreservation by slow freezing/vitrification-i.e., the cryopreservative medium. The cryopreservative medium is typically specified as a dilution of a concentrated cryoprotectant, such as dimethylsulfoxide or glycerol, in cell culture medium that is often combined with serum or another source of necessary growth factors. The present work is devoted to a computational tool for determining the composition of a cryopreservative medium that can be combined with dissociated fhNSC resuspended in a certain volume of culture medium to achieve the criterion of stoichiometric dilution of cryoprotectant favorable to cell viability in the final mixture of cryopreservative medium and cells. © 2021 Wiley Periodicals LLC. Basic Protocol: Culture and passage of fhNSC, counting of enzymatically dissociated fhNSC, and quantitative formulation of cryomedium Alternate Protocol: Procedure when cell medium is not added to the cryomedium.
- Published
- 2021
24. Reversal of Surfactant Protein B Deficiency in Patient Specific Human Induced Pluripotent Stem Cell Derived Lung Organoids by Gene Therapy
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Evan Y. Snyder, Jinxia Wang, Alicia M. Winquist, Irene Tseu, Sharareh Shojaie, Daochun Luo, Neal H. Nathan, Martin Post, and Sandra L Leibel
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Genetic Markers ,Cellular differentiation ,Genetic enhancement ,Organogenesis ,Green Fluorescent Proteins ,Induced Pluripotent Stem Cells ,lcsh:Medicine ,Biology ,Lamellar granule ,Pulmonary Alveolar Proteinosis ,Article ,03 medical and health sciences ,0302 clinical medicine ,Pluripotent stem cells ,Surfactant Protein B Deficiency ,Organoid ,Humans ,Induced pluripotent stem cell ,lcsh:Science ,Lung ,030304 developmental biology ,0303 health sciences ,Respiratory tract diseases ,Multidisciplinary ,Pulmonary Surfactant-Associated Protein B ,Mesenchymal stem cell ,Lentivirus ,lcsh:R ,Cell Differentiation ,Epithelial Cells ,Genetic Therapy ,respiratory system ,Fibroblasts ,3. Good health ,Organoids ,Pulmonary Alveoli ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,lcsh:Q - Abstract
Surfactant protein B (SFTPB) deficiency is a fatal disease affecting newborn infants. Surfactant is produced by alveolar type II cells which can be differentiated in vitro from patient specific induced pluripotent stem cell (iPSC)-derived lung organoids. Here we show the differentiation of patient specific iPSCs derived from a patient with SFTPB deficiency into lung organoids with mesenchymal and epithelial cell populations from both the proximal and distal portions of the human lung. We alter the deficiency by infecting the SFTPB deficient iPSCs with a lentivirus carrying the wild type SFTPB gene. After differentiating the mutant and corrected cells into lung organoids, we show expression of SFTPB mRNA during endodermal and organoid differentiation but the protein product only after organoid differentiation. We also show the presence of normal lamellar bodies and the secretion of surfactant into the cell culture medium in the organoids of lentiviral infected cells. These findings suggest that a lethal lung disease can be targeted and corrected in a human lung organoid model in vitro.
- Published
- 2019
25. Generation of 3D Whole Lung Organoids from Induced Pluripotent Stem Cells for Modeling Lung Developmental Biology and Disease
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Evan Y. Snyder, Alicia M. Winquist, Rachael N. McVicar, and Sandra L Leibel
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Organogenesis ,Mesenchyme ,Cellular differentiation ,General Chemical Engineering ,Induced Pluripotent Stem Cells ,Cell Culture Techniques ,Biology ,General Biochemistry, Genetics and Molecular Biology ,medicine ,Humans ,Progenitor cell ,Induced pluripotent stem cell ,Lung ,Tissue Engineering ,General Immunology and Microbiology ,General Neuroscience ,Endoderm ,Mesenchymal stem cell ,Cell Differentiation ,respiratory system ,Embryonic stem cell ,Cell biology ,Organoids ,medicine.anatomical_structure ,Cell culture ,Alveolar Epithelial Cells ,Developmental biology ,Developmental Biology - Abstract
Human lung development and disease has been difficult to study due to the lack of biologically relevant in vitro model systems. Human induced pluripotent stem cells (hiPSCs) can be differentiated stepwise into 3D multicellular lung organoids, made of both epithelial and mesenchymal cell populations. We recapitulate embryonic developmental cues by temporally introducing a variety of growth factors and small molecules to efficiently generate definitive endoderm, anterior foregut endoderm, and subsequently lung progenitor cells. These cells are then embedded in growth factor reduced (GFR)-basement membrane matrix medium, allowing them to spontaneously develop into 3D lung organoids in response to external growth factors. These whole lung organoids (WLO) undergo early lung developmental stages including branching morphogenesis and maturation after exposure to dexamethasone, cyclic AMP and isobutylxanthine. WLOs possess airway epithelial cells expressing the markers KRT5 (basal), SCGB3A2 (club) and MUC5AC (goblet) as well as alveolar epithelial cells expressing HOPX (alveolar type I) and SP-C (alveolar type II). Mesenchymal cells are also present, including smooth muscle actin (SMA), and platelet-derived growth factor receptor A (PDGFRα). iPSC derived WLOs can be maintained in 3D culture conditions for many months and can be sorted for surface markers to purify a specific cell population. iPSC derived WLOs can also be utilized to study human lung development, including signaling between the lung epithelium and mesenchyme, to model genetic mutations on human lung cell function and development, and to determine the cytotoxicity of infective agents.
- Published
- 2021
26. Chemical mutagenesis of a GPCR ligand: Detoxifying 'inflammo-attraction' to direct therapeutic stem cell migration
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Dustin R. Wakeman, Thomas N. Seyfried, Edward B. Han, Ziwei Huang, Rodolfo Gonzalez, Walter L. Niles, Yongmei Feng, Milton H. Hamblin, Maocai Yan, Runquan Zhang, Jing An, Evan Y. Snyder, Juan Wang, Srinivas Duggineni, Jean Pyo Lee, Xiao Fang, Yan Xu, David A. Wenger, Richard L. Sidman, Yinsong Zhu, and Justin Chen
- Subjects
Central Nervous System ,Chemokine ,Receptors, CXCR4 ,medicine.medical_treatment ,Cell ,Induced Pluripotent Stem Cells ,Ligands ,CXCR4 ,Neural Stem Cells ,Cell Movement ,medicine ,Humans ,Induced pluripotent stem cell ,Inflammation ,Neurons ,Multidisciplinary ,biology ,Chemistry ,neurodegeneration ,Neurodegenerative Diseases ,Cell Biology ,Biological Sciences ,Neural stem cell ,Chemokine CXCL12 ,Cell biology ,human induced pluripotent stem cells ,medicine.anatomical_structure ,Cytokine ,Mutagenesis ,Astrocytes ,biology.protein ,Stem cell ,homing ,Homing (hematopoietic) ,Protein Binding - Abstract
Significance While inflammatory chemokines, constitutively produced by pathologic regions, are pivotal for attracting reparative stem cells, one would certainly not want to further “inflame” a diseased brain by instilling such molecules. Exploiting the fact that receptors for such cytokines (G protein-coupled receptors [GPCR]) possess two “pockets”—one for binding, the other for signaling—we created a synthetic GPCR-agonist that maximizes interaction with the former and narrows that with the latter. Homing is robust with no inflammation. The peptide successfully directed the integration of human induced pluripotent stem cell derivatives (known to have muted migration) in a model of a prototypical neurodegenerative condition, ameliorating symptomatology., A transplanted stem cell’s engagement with a pathologic niche is the first step in its restoring homeostasis to that site. Inflammatory chemokines are constitutively produced in such a niche; their binding to receptors on the stem cell helps direct that cell’s “pathotropism.” Neural stem cells (NSCs), which express CXCR4, migrate to sites of CNS injury or degeneration in part because astrocytes and vasculature produce the inflammatory chemokine CXCL12. Binding of CXCL12 to CXCR4 (a G protein-coupled receptor, GPCR) triggers repair processes within the NSC. Although a tool directing NSCs to where needed has been long-sought, one would not inject this chemokine in vivo because undesirable inflammation also follows CXCL12–CXCR4 coupling. Alternatively, we chemically “mutated” CXCL12, creating a CXCR4 agonist that contained a strong pure binding motif linked to a signaling motif devoid of sequences responsible for synthetic functions. This synthetic dual-moity CXCR4 agonist not only elicited more extensive and persistent human NSC migration and distribution than did native CXCL 12, but induced no host inflammation (or other adverse effects); rather, there was predominantly reparative gene expression. When co-administered with transplanted human induced pluripotent stem cell-derived hNSCs in a mouse model of a prototypical neurodegenerative disease, the agonist enhanced migration, dissemination, and integration of donor-derived cells into the diseased cerebral cortex (including as electrophysiologically-active cortical neurons) where their secreted cross-corrective enzyme mediated a therapeutic impact unachieved by cells alone. Such a “designer” cytokine receptor-agonist peptide illustrates that treatments can be controlled and optimized by exploiting fundamental stem cell properties (e.g., “inflammo-attraction”).
- Published
- 2020
27. Nna1 gene deficiency triggers Purkinje neuron death by tubulin hyperglutamylation and ER dysfunction
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Evan Y. Snyder, Renata Pasqualini, Wadih Arap, Fenny H.Tang Tang, Richard L. Sidman, and Jianxue Li
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Male ,0301 basic medicine ,animal structures ,Glutamine ,medicine.medical_treatment ,Necroptosis ,Purkinje cell ,Apoptosis ,Substantia nigra ,Biology ,Endoplasmic Reticulum ,Mice ,Purkinje Cells ,03 medical and health sciences ,0302 clinical medicine ,GTP-Binding Proteins ,Tubulin ,Microtubule ,medicine ,Animals ,Neurodegeneration ,Peptide Synthases ,Mice, Knockout ,Neurons ,General Medicine ,medicine.disease ,Granule cell ,Serine-Type D-Ala-D-Ala Carboxypeptidase ,Cell biology ,Mice, Inbred C57BL ,Phenotype ,030104 developmental biology ,medicine.anatomical_structure ,nervous system ,030220 oncology & carcinogenesis ,Mutation ,Unfolded protein response ,Medicine ,Female ,Axotomy ,Research Article ,Neuroscience - Abstract
Posttranslational glutamylation/deglutamylation balance in tubulins influences dendritic maturation and neuronal survival of cerebellar Purkinje neurons (PNs). PNs and some additional neuronal types degenerate in several spontaneous, independently occurring Purkinje cell degeneration (pcd) mice featuring mutant neuronal nuclear protein induced by axotomy (Nna1), a deglutamylase gene. This defective deglutamylase allows glutamylases to form hyperglutamylated tubulins. In pcd, all PNs die during postnatal “adolescence.” Neurons in some additional brain regions also die, mostly later than PNs. We show in laser capture microdissected single PNs, in cerebellar granule cell neuronal clusters, and in dissected hippocampus and substantia nigra that deglutamase mRNA and protein were virtually absent before pcd PNs degenerated, whereas glutaminase mRNA and protein remained normal. Hyperglutamylated microtubules and dimeric tubulins accumulated in pcd PNs and were involved in pcd PN death by glutamylase/deglutamylase imbalance. Importantly, treatment with a microtubule depolymerizer corrected the glutamylation/deglutamylation ratio, increasing PN survival. Further, before onset of neuronal death, pcd PNs displayed prominent basal polylisosomal masses rich in ER. We propose a “seesaw” metamorphic model summarizing mutant Nna1-induced tubulin hyperglutamylation, the pcd’s PN phenotype, and report that the neuronal disorder involved ER stress, unfolded protein response, and protein synthesis inhibition preceding PN death by apoptosis/necroptosis., Purkinje cell degeneration is due to ER stress, unfolded protein response, and protein synthesis inhibition preceding Purkinje neuron death by apoptosis/necroptosis.
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- 2020
28. Deriving Keratinocyte Progenitor Cells and Keratinocytes from Human‐Induced Pluripotent Stem Cells
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Michel R Ibrahim, Hamza Abdel-Raouf, Walid Medhat, Evan Y. Snyder, and Hasan El-Fakahany
- Subjects
Keratinocytes ,0301 basic medicine ,Keratin 14 ,Induced Pluripotent Stem Cells ,Cell Culture Techniques ,Biology ,Transfection ,Cell Line ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Keratin ,medicine ,Animals ,Humans ,Progenitor cell ,Induced pluripotent stem cell ,Embryoid Bodies ,Skin ,chemistry.chemical_classification ,integumentary system ,Cell Differentiation ,DNA ,Cell Biology ,General Medicine ,Fibroblasts ,Cellular Reprogramming ,Hair follicle ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Stem cell ,Keratinocyte ,Reprogramming ,030217 neurology & neurosurgery ,Plasmids ,Developmental Biology - Abstract
Skin or hair loss (alopecia) may occur due to a wide variety of causes ranging from trauma to pathological processes including acquired or congenital causes. It would be ideal to replace them with immunologically compatible cells to avoid potentially exacerbating the condition. Deriving the replacement cells from human-induced pluripotent stem cells (hiPSCs) allows for sufficient scale up and using hiPSCs as the choice of human pluripotent stem cells (hPSC) will ensure immunocompatibility. Here we offer a protocol for differentiating hiPSCs into keratinocyte progenitor cells (KPC) and keratinocytes employing all-trans retinoic acid (ATRA) and L-ascorbic acid, (L-AA), bone morphogenic protein-4 (BMP4), and epidermal growth factor (EGF). We observed that the hiPSC-derived KPCs express the same panel of markers as primary hair follicle bulge stem cells (HFBSCs), including CD200, integrin α-6 (ITGA6), integrin β-1 (ITGB1), the transcription factor P63, keratin 15 (KRT15), and keratin 19 (KRT19). If permitted to differentiate further, the hiPSC-derived KPC lose CD200 expression and rather come to express keratin 14 (KRT14) indicating emergence of more mature terminally-differentiated keratinocytes. The HFBSCs are transplantable for hair follicle (HF) restoration, and the keratinocytes may be transplantable for therapy for large burns or ulcers. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Reprogramming of normal human skin fibroblasts into normal hiPSCs using episomal DNA cocktail Basic Protocol 2: Differentiation of hiPSCs into KPCs and keratinocytes Alternate Protocol 2: EBS formation protocol using AggreWell™ plates (Antonchuk, 2013) Support Protocol 1: Passage hiPSC-KPC Support Protocol 2: Immunocytochemistry (ICC) Support Protocol 3: Immunofluorescence staining of cells for flow cytometry (FC).
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- 2020
29. Generation of Complete Multi-Cell Type Lung Organoids From Human Embryonic and Patient-Specific Induced Pluripotent Stem Cells for Infectious Disease Modeling and Therapeutics Validation
- Author
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Rachael N. McVicar, Alicia M. Winquist, Evan Y. Snyder, Sandra L Leibel, and Walter D. Niles
- Subjects
0301 basic medicine ,Patient-Specific Modeling ,ciliated cells ,Cellular differentiation ,lung organoid ,Human Embryonic Stem Cells ,Cell Culture Techniques ,smooth muscle ,0302 clinical medicine ,Protocol ,human pluripotent stem cells ,Induced pluripotent stem cell ,Lung ,Respiratory Tract Infections ,pulmonary disease modeling ,Cell Differentiation ,General Medicine ,differentiation ,respiratory system ,Cell biology ,Organoids ,medicine.anatomical_structure ,Endoderm ,Ls30 ,Coronavirus Infections ,Cell type ,surfactant ,Induced Pluripotent Stem Cells ,Pneumonia, Viral ,Biology ,Models, Biological ,Time-Lapse Imaging ,03 medical and health sciences ,Betacoronavirus ,medicine ,Humans ,Ls40 ,Progenitor cell ,endoderm ,Pandemics ,lung development ,SARS-CoV-2 ,Mesenchymal stem cell ,COVID-19 ,type 2 alveolar cells ,Cell Biology ,SARS‐CoV‐2 viral infection ,Embryonic stem cell ,Ls90 ,030104 developmental biology ,Cell culture ,030217 neurology & neurosurgery ,Developmental Biology ,type 1 alveolar cells - Abstract
The normal development of the pulmonary system is critical to transitioning from placental‐dependent fetal life to alveolar‐dependent newborn life. Human lung development and disease have been difficult to study due to the lack of an in vitro model system containing cells from the large airways and distal alveolus. This article describes a system that allows human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) to differentiate and form three‐dimensional (3D) structures that emulate the development, cytoarchitecture, and function of the lung (“organoids”), containing epithelial and mesenchymal cell populations, and including the production of surfactant and presence of ciliated cells. The organoids can also be invested with mesoderm derivatives, differentiated from the same human pluripotent stem cells, such as alveolar macrophages and vasculature. Such lung organoids may be used to study the impact of environmental modifiers and perturbagens (toxins, microbial or viral pathogens, alterations in microbiome) or the efficacy and safety of drugs, biologics, and gene transfer. © 2020 Wiley Periodicals LLC. Basic Protocol: hESC/hiPSC dissection, definitive endoderm formation, and lung progenitor cell induction
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- 2020
30. Discovery of suppressors of CRMP2 phosphorylation reveals compounds that mimic the behavioral effects of lithium on amphetamine-induced hyperlocomotion
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Deepak Mani, Cameron D. Pernia, Steven D. Sheridan, Jasmin Lalonde, Joshua A. Bishop, Brian T. D. Tobe, Debasis Patnaik, Namrata D. Udeshi, Stephen J. Haggarty, Steven A. Carr, Lucius L Xuan, Irina N. Gaisina, Evan Y. Snyder, Sarah R. Blumenthal, Daniel P Zolg, Wen-Ning Zhao, Amanda J. Roberts, and Iren Kurtser
- Subjects
0301 basic medicine ,Bipolar Disorder ,Lithium (medication) ,Induced Pluripotent Stem Cells ,Lithium ,Predictive markers ,Molecular neuroscience ,Article ,lcsh:RC321-571 ,Mice ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Stable isotope labeling by amino acids in cell culture ,medicine ,Animals ,Humans ,Phosphorylation ,Kinase activity ,Protein kinase A ,Induced pluripotent stem cell ,Amphetamine ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,Biological Psychiatry ,Chemistry ,3. Good health ,Cell biology ,Psychiatry and Mental health ,030104 developmental biology ,Lithium Compounds ,Collapsin response mediator protein family ,030217 neurology & neurosurgery ,medicine.drug - Abstract
The effective treatment of bipolar disorder (BD) represents a significant unmet medical need. Although lithium remains a mainstay of treatment for BD, limited knowledge regarding how it modulates affective behavior has proven an obstacle to discovering more effective mood stabilizers with fewer adverse side effects. One potential mechanism of action of lithium is through inhibition of the serine/threonine protein kinase GSK3β, however, relevant substrates whose change in phosphorylation may mediate downstream changes in neuroplasticity remain poorly understood. Here, we used human induced pluripotent stem cell (hiPSC)-derived neuronal cells and stable isotope labeling by amino acids in cell culture (SILAC) along with quantitative mass spectrometry to identify global changes in the phosphoproteome upon inhibition of GSK3α/β with the highly selective, ATP-competitive inhibitor CHIR-99021. Comparison of phosphorylation changes to those induced by therapeutically relevant doses of lithium treatment led to the identification of collapsin response mediator protein 2 (CRMP2) as being highly sensitive to both treatments as well as an extended panel of structurally distinct GSK3α/β inhibitors. On this basis, a high-content image-based assay in hiPSC-derived neurons was developed to screen diverse compounds, including FDA-approved drugs, for their ability to mimic lithium’s suppression of CRMP2 phosphorylation without directly inhibiting GSK3β kinase activity. Systemic administration of a subset of these CRMP2-phosphorylation suppressors were found to mimic lithium’s attenuation of amphetamine-induced hyperlocomotion in mice. Taken together, these studies not only provide insights into the neural substrates regulated by lithium, but also provide novel human neuronal assays for supporting the development of mechanism-based therapeutics for BD and related neuropsychiatric disorders.
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- 2020
31. Deciphering the Systems Architecture of the Brain Using Molecular Can Openers
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Stephen J. Haggarty, Brian T. D. Tobe, Yoshio Goshima, Cameron D. Pernia, Evan Y. Snyder, Neal H. Nathan, and Richard L. Sidman
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Modern medicine ,Human patient ,Systems architecture ,Context (language use) ,Business ,Induced pluripotent stem cell ,Data science ,Healthcare providers - Abstract
Throughout much of human and medical history the brain has been a black box opaque to scientific investigation for a myriad of reasons ranging from societal to logistical barriers. Compounding this opacity, the biological effects of ecologically sourced medicines that early cultures, and many current societies, prescribed were inherently elusive due to technological hurdles. However, in modern medicine and science, these challenges are now surmountable. Healthy and diseased human patient neurons are now more accessible, as noninvasively collected somatic cells, such as skin cells, can be reprogrammed into induced pluripotent stem cells (iPSCs) and then differentiated into neurons in vitro. Due to this contemporary ability to grow patient neurons in a dish, in conjunction with tractable animal models, functional agents can now be deployed to pry open the black box and gain insight into the construction of the brain and the pathophysiology of disease. This chapter will use case studies of peer-reviewed and published research to demonstrate the explanatory power of functional agents that serve as ‘molecular can openers’, such as in the case of the functional agent lithium in the context of bipolar disorder. This chapter will also consider the value iPSCs provide to research and development and offer thoughts on the business and translational aspects of this technology as well as share contemplations on the future of this field. This chapter aims to depict how collaboration between basic science laboratories, clinical research institutions, biotechnology businesses, regulatory apparatuses and healthcare providers can not only improve health and profit outcomes but also improve performance along the innovation S-curve.
- Published
- 2020
32. List of Contributors
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Julie G. Allickson, Rafael G. Amado, Usman Azam, Frank Barry, Irene Aldecoa Bilbao, Gwendolyn K. Binder, Justin L. Black, Karin Blumer, Eugene P. Brandon, David A. Brindley, William Murray Burns, Francesca Capone, Guillaume Carmona, Michael Chaffman, William Chou, Zhanfeng Cui, Kevin A. D’Amour, Nathan J. Dowden, Bernd Eschgfaeller, Eric Faulkner, Maria Cristina Galli, Yoshio Goshima, Stephen J. Haggarty, Celine-Lea Halioua-Haubold, Kristen Harrington-Smith, Darren Hickerson, William Ho, Johan Hyllner, Natasha Iyer, Marieke Jansen, Etienne Jousseaume, Sven Kili, Antoinette F. Konski, Lawrence S. Lamb, Ching Lam, Jaeseung Lim, Chi Lo, Thomas Malcolm, Almudena Martinez-Fernandez, Todd Meinecke, Edward Meinert, Michael C. Milone, David A. Mitchell, Niraj Muni, Filomena Nappi, Neal H. Nathan, Emanuele Ostuni, Marianthi Papakosta, Simona Paratore, Vincenzo Lorenzo Pascali, Cameron D. Pernia, Benjamin Philipson, Fiona Phillips, James B. Phillips, Nasib Qureshi, Pietro Refolo, Moira Ringo, Matthew Robson, Crystal Ruff, Manuel Schmidt, Matthias Schroff, Michael J. Scott, Michaela Sharpe, Richard L. Sidman, John D. Sinden, Devyn M. Smith, Evan Y. Snyder, Sita Somara, Antonio Gioacchino Spagnolo, Daryl S. Spinner, Kanaka Sridharan, Michael Sullivan, Keith Thompson, Brian T.D. Tobe, Alain A. Vertès, Michael D. West, Cynthia Wilkins-Port, Burghardt Wittig, Aidong Yang, Jie Zhang, and Mark C. Zimmerman
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- 2020
33. Alteration of the protein levels and/or posttranslational modification of CRMP 2 (Collapsin Response Mediator Protein 2) in the peripheral blood samples from young schizophrenia patients
- Author
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Nomoto, Munetaka, primary, Yamashita, Naoya, additional, Jitsuki-Takahashi, Aoi, additional, Nakamura, Haruko, additional, Makihara, Hiroko, additional, Nakamura, Fumio, additional, Evan Y, Snyder, additional, Hirayasu, Yoshio, additional, and Goshima, Yoshio, additional
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- 2020
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34. 'Reprogram Enablement' as an Assay for Identifying Early Oncogenic Pathways by Their Ability to Allow Neoplastic Cells to Reacquire an Epiblast State
- Author
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Rachael N. McVicar, Ryan C. Gimple, Yanjun Kong, Yang Liu, Andrew Hodges, Weiwei Zhan, Evan Y. Snyder, and Jun Yin
- Subjects
cancer stem cells ,0301 basic medicine ,Transcription, Genetic ,Somatic cell ,Carcinogenesis ,medicine.disease_cause ,Thyroid Carcinoma, Anaplastic ,Biochemistry ,Sendai virus ,Epigenesis, Genetic ,reprogram enablement ,0302 clinical medicine ,Neoplasms ,organoids ,epiblast ,Cell Differentiation ,human embryonic stem cells ,Cellular Reprogramming ,Gene Expression Regulation, Neoplastic ,Phenotype ,Neoplastic Stem Cells ,Biological Assay ,Reprogramming ,Germ Layers ,tumor ,Induced Pluripotent Stem Cells ,Down-Regulation ,Biology ,Article ,03 medical and health sciences ,oncogenesis ,Cancer stem cell ,Genetics ,medicine ,cancer ,Humans ,Epigenetics ,Thyroid Neoplasms ,reprogramming ,Cell Biology ,DNA Methylation ,Embryonic stem cell ,human induced pluripotent stem cells ,030104 developmental biology ,Epiblast ,Cancer cell ,Mutation ,Cancer research ,ras Proteins ,030217 neurology & neurosurgery ,RAS ,Developmental Biology - Abstract
Summary One approach to understanding how tissue-specific cancers emerge is to determine the requirements for “reprograming” such neoplastic cells back to their developmentally normal primordial pre-malignant epiblast-like pluripotent state and then scrutinizing their spontaneous reconversion to a neoplasm, perhaps rendering salient the earliest pivotal oncogenic pathway(s) (before other aberrations accumulate in the adult tumor). For the prototypical malignancy anaplastic thyroid carcinoma (ATC), we found that tonic RAS reduction was obligatory for reprogramming cancer cells to a normal epiblast-emulating cells, confirmed by changes in their transcriptomic and epigenetic profiles, loss of neoplastic behavior, and ability to derive normal somatic cells from their “epiblast organoids.” Without such suppression, ATCs re-emerged from the clones. Hence, for ATC, RAS inhibition was its “reprogram enablement” (RE) factor. Each cancer likely has its own RE factor; identifying it may illuminate pre-malignant risk markers, better classifications, therapeutic targets, and tissue-specification of a previously pluripotent, now neoplastic, cell., Highlights • The factors for reprogramming a cancer cell to an epiblast-like cell can be assayed • “Reprogram enablement” can yield insights into the earliest pivotal oncogenic steps • For anaplastic thyroid carcinoma, RAS inhibition was obligatory for reprograming • Each tissue-specific cancer will have its own reprogramming enablement requirement, In this article, Snyder and colleagues show that the minimal essential factor(s) needed to convert a cancer cell back to its normal, pre-malignant, epiblast-like pluripotent state and then back again into a neoplastic cell—its “reprogram enablement factor”—may provide insights into the earliest pivotal steps in that cancer's tissue-specific oncogenesis, as well as its early diagnosis and treatment.
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- 2019
35. Stem Cell Composition of Umbilical Cord Blood Following Milking Compared with Delayed Clamping of the Cord Appears Better Suited for Promoting Hematopoiesis
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Evan Y. Snyder, Anthony J. Orona, Yang Liu, Jason M. Konop, Donlaong Wu, Anup Katheria, Ramina Amino, and Elizabeth Kim
- Subjects
Pathology ,medicine.medical_specialty ,Blood Specimen Collection ,Cord ,Time Factors ,business.industry ,Anemia ,Mesenchymal stem cell ,Irradiated bone ,Mesenchymal Stem Cells ,medicine.disease ,Fetal Blood ,Hematopoietic Stem Cells ,Umbilical cord ,Constriction ,Milking ,Hematopoiesis ,Haematopoiesis ,Mice ,medicine.anatomical_structure ,Pediatrics, Perinatology and Child Health ,Medicine ,Animals ,Stem cell ,business - Abstract
In comparing placental transfusion strategies, blood obtained from an umbilical cord that has been "milked" vs one in which clamping was simply delayed contains mesenchymal stromal cells in addition to solely hematopoietic stem cells, a composition more favorable for hematopoiesis, as suggested by its superior rescue of lethally irradiated bone marrow-depleted mice.
- Published
- 2019
36. The Developmental & Molecular Requirements for Ensuring that Human Pluripotent Stem Cell-Derived Hair Follicle Bulge Stem Cells Have Acquired Competence for Hair Follicle Generation Following Transplantation
- Author
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Evan Y. Snyder, Hamza Abdel-Raouf, Hasan El-Fakahany, Walid Medhat, and Michel R Ibrahim
- Subjects
keratinocytes ,Pluripotent Stem Cells ,CD200 glycoprotein ,hair follicle replacement ,Keratin 14 ,human induced pluripotent stem cells (hiPSCs) ,Biomedical Engineering ,hair follicle bulge stem cells (HFBSCs) ,Biology ,Regenerative Medicine ,Keratin 18 ,medicine ,Humans ,Progenitor cell ,Induced pluripotent stem cell ,Transplantation ,Cell Biology ,Hair follicle ,Cell biology ,Keratin 5 ,medicine.anatomical_structure ,integrins ,Medicine ,Original Article ,Stem cell ,human pluripotent stem cells (hPSCs) ,Hair Follicle ,human embryonic stem cells (hESCs) - Abstract
When using human induced pluripotent stem cells (hiPSCs) to achieve hair follicle (HF) replacement, we found it best to emulate the earliest fundamental developmental processes of gastrulation, ectodermal lineage commitment, and dermogenesis. Viewing hiPSCs as a model of the epiblast, we exploited insights from mapping the dynamic up- and down-regulation of the developmental molecules that determine HF lineage in order to ascertain the precise differentiation stage and molecular requirements for grafting HF-generating progenitors. To yield an integrin-dependent lineage like the HF in vivo, we show that hiPSC derivatives should co-express, just prior to transplantation, the following combination of markers: integrins α6 and β1 and the glycoprotein CD200 on their surface; and, intracellularly, the epithelial marker keratin 18 and the hair follicle bulge stem cell (HFBSC)-defining molecules transcription factor P63 and the keratins 15 and 19. If the degree of trichogenic responsiveness indicated by the presence of these molecules is not achieved (they peak on Days 11-18 of the protocol), HF generation is not possible. Conversely, if differentiation of the cells is allowed to proceed beyond the transient intermediate progenitor state represented by the HFBSC, and instead cascades to their becoming keratin 14+ keratin 5+ CD200– keratinocytes (Day 25), HF generation is equally impossible. We make the developmental case for transplanting at Day 16-18 of differentiation—the point at which the hiPSCs have lost pluripotency, have attained optimal expression of HFBSC markers, have not yet experienced downregulation of key integrins and surface glycoproteins, have not yet started expressing keratinocyte-associated molecules, and have sufficient proliferative capacity to allow a well-populated graft. This panel of markers may be used for isolating (by cytometry) HF-generating derivatives away from cell types unsuited for this therapy as well as for identifying trichogenic drugs.
- Published
- 2021
37. Quantitative Analysis of Human Pluripotency and Neural Specification by In-Depth (Phospho)Proteomic Profiling
- Author
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Brian T. D. Tobe, Alicia M. Winquist, Laurence M. Brill, Gustavo J. Gutierrez, Andrew Crain, Stuart A. Lipton, Junjie Hou, Jennifer Choy, Kutbuddin S. Doctor, Maria Talantova, Evan Y. Snyder, Esther La Monaca, Dieter A. Wolf, Xiayu Huang, David Horn, Ilyas Singec, and Biology
- Subjects
0301 basic medicine ,Proteomics ,Proteome ,Cellular differentiation ,Stem Cell Research - Embryonic - Non-Human ,Regenerative Medicine ,Biochemistry ,0302 clinical medicine ,Induced pluripotent stem cell ,lcsh:QH301-705.5 ,Cells, Cultured ,Neurons ,lcsh:R5-920 ,Cultured ,Cell Differentiation ,Neural stem cell ,3. Good health ,Cell biology ,Gene Knockdown Techniques ,Stem cell ,lcsh:Medicine (General) ,Neural development ,Signal Transduction ,Resource ,Pluripotent Stem Cells ,Cells ,1.1 Normal biological development and functioning ,Clinical Sciences ,Biology ,03 medical and health sciences ,Underpinning research ,Genetics ,Animals ,Humans ,Cell Lineage ,Stem Cell Research - Embryonic - Human ,Transplantation ,Proteomic Profiling ,Gene Expression Profiling ,Neurosciences ,Computational Biology ,Cell Biology ,Stem Cell Research ,Phosphoproteins ,Embryonic stem cell ,030104 developmental biology ,lcsh:Biology (General) ,Generic health relevance ,Biochemistry and Cell Biology ,Transcriptome ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Summary Controlled differentiation of human embryonic stem cells (hESCs) can be utilized for precise analysis of cell type identities during early development. We established a highly efficient neural induction strategy and an improved analytical platform, and determined proteomic and phosphoproteomic profiles of hESCs and their specified multipotent neural stem cell derivatives (hNSCs). This quantitative dataset (nearly 13,000 proteins and 60,000 phosphorylation sites) provides unique molecular insights into pluripotency and neural lineage entry. Systems-level comparative analysis of proteins (e.g., transcription factors, epigenetic regulators, kinase families), phosphorylation sites, and numerous biological pathways allowed the identification of distinct signatures in pluripotent and multipotent cells. Furthermore, as predicted by the dataset, we functionally validated an autocrine/paracrine mechanism by demonstrating that the secreted protein midkine is a regulator of neural specification. This resource is freely available to the scientific community, including a searchable website, PluriProt., Highlights • Controlled neural induction produces pure cultures of PAX6+ neural stem cells • Most comprehensive (phospho)proteome mapping in pluripotent and multipotent cells • Prediction and validation of midkine as regulator of neural lineage commitment • Searchable and publicly available website presenting (phospho)proteomic dataset, Snyder, Brill, Singec, and colleagues demonstrate detailed analysis of human pluripotency and controlled neural lineage entry by using quantitative label-free (phospho)proteomics. The accuracy of the large dataset (13,000 proteins; 60,000 non-redundant phosphorylation sites) allows precise characterization and comparison of pluripotent and multipotent “stemness.” Functional follow-up experiments validate that the understudied protein midkine controls neuralization of hESCs.
- Published
- 2016
38. Reduced-representation Phosphosignatures Measured by Quantitative Targeted MS Capture Cellular States and Enable Large-scale Comparison of Drug-induced Phenotypes
- Author
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Evan Y. Snyder, Adam Officer, Xiaodong Lu, Jacob D. Jaffe, Eric Kuhn, Jinal Patel, Jana W. Qiao, Lola Fagbami, Steven A. Carr, Roger Hu, Desiree Davison, Lindsay K. Pino, Caitlin M. Feeney, Aravind Subramanian, Susan E. Abbatiello, Jianxue Li, Richard L. Sidman, Jennifer G. Abelin, Daniel D. Lam, and Amanda L. Creech
- Subjects
Proteomics ,0301 basic medicine ,Analyte ,Quantitative proteomics ,Computational biology ,Biochemistry ,Analytical Chemistry ,Small Molecule Libraries ,Mice ,03 medical and health sciences ,0302 clinical medicine ,High-Throughput Screening Assays ,Gene expression ,Animals ,Humans ,Molecular Biology ,Cells, Cultured ,Embryonic Stem Cells ,Chemistry ,Phosphopeptide ,Research ,Phosphoproteomics ,Proteolytic enzymes ,Phosphoproteins ,Small molecule ,030104 developmental biology ,030220 oncology & carcinogenesis ,MCF-7 Cells ,Signal Transduction - Abstract
Profiling post-translational modifications represents an alternative dimension to gene expression data in characterizing cellular processes. Many cellular responses to drugs are mediated by changes in cellular phosphosignaling. We sought to develop a common platform on which phosphosignaling responses could be profiled across thousands of samples, and created a targeted MS assay that profiles a reduced-representation set of phosphopeptides that we show to be strong indicators of responses to chemical perturbagens. To develop the assay, we investigated the coordinate regulation of phosphosites in samples derived from three cell lines treated with 26 different bioactive small molecules. Phosphopeptide analytes were selected from these discovery studies by clustering and picking 1 to 2 proxy members from each cluster. A quantitative, targeted parallel reaction monitoring assay was developed to directly measure 96 reduced-representation probes. Sample processing for proteolytic digestion, protein quantification, peptide desalting, and phosphopeptide enrichment have been fully automated, making possible the simultaneous processing of 96 samples in only 3 days, with a plate phosphopeptide enrichment variance of 12%. This highly reproducible process allowed ∼95% of the reduced-representation phosphopeptide probes to be detected in ∼200 samples. The performance of the assay was evaluated by measuring the probes in new samples generated under treatment conditions from discovery experiments, recapitulating the observations of deeper experiments using a fraction of the analytical effort. We measured these probes in new experiments varying the treatments, cell types, and timepoints to demonstrate generalizability. We demonstrated that the assay is sensitive to disruptions in common signaling pathways (e.g. MAPK, PI3K/mTOR, and CDK). The high-throughput, reduced-representation phosphoproteomics assay provides a platform for the comparison of perturbations across a range of biological conditions, suitable for profiling thousands of samples. We believe the assay will prove highly useful for classification of known and novel drug and genetic mechanisms through comparison of phosphoproteomic signatures.
- Published
- 2016
39. A Biomarker for Predicting Responsiveness to Stem Cell Therapy Based on Mechanism-of-Action: Evidence from Cerebral Injury
- Author
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Dustin R. Wakeman, Nirmalya Ghosh, Amy Plaia, Nejmi Dilmac, Walter D. Niles, Richard E. Hartman, Ruslan Nuryyev, Evan Y. Snyder, Neal H. Nathan, Melissa S. Dulcich, Alena Yusof, Richard L. Sidman, Beatriz Tone, Stephen Ashwal, Janessa B Law, Cameron D. Pernia, and Andre Obenaus
- Subjects
0301 basic medicine ,medicine.medical_treatment ,Regenerative Medicine ,Regenerative medicine ,Neuroprotection ,General Biochemistry, Genetics and Molecular Biology ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Medicine ,medicine.diagnostic_test ,business.industry ,Penumbra ,Magnetic resonance imaging ,Stem-cell therapy ,Neural stem cell ,Rats ,Biomarker (cell) ,Transplantation ,Disease Models, Animal ,030104 developmental biology ,Brain Injuries ,business ,Neuroscience ,Biomarkers ,030217 neurology & neurosurgery ,Stem Cell Transplantation - Abstract
To date, no stem cell therapy has been directed to specific recipients-and, conversely, withheld from others-based on a clinical or molecular profile congruent with that cell's therapeutic mechanism-of-action (MOA) for that condition. We address this challenge preclinically with a prototypical scenario: human neural stem cells (hNSCs) against perinatal/neonatal cerebral hypoxic-ischemic injury (HII). We demonstrate that a clinically translatable magnetic resonance imaging (MRI) algorithm, hierarchical region splitting, provides a rigorous, expeditious, prospective, noninvasive "biomarker" for identifying subjects with lesions bearing a molecular profile indicative of responsiveness to hNSCs' neuroprotective MOA. Implanted hNSCs improve lesional, motor, and/or cognitive outcomes only when there is an MRI-measurable penumbra that can be forestalled from evolving into necrotic core; the core never improves. Unlike the core, a penumbra is characterized by a molecular profile associated with salvageability. Hence, only lesions characterized by penumbral > core volumes should be treated with cells, making such measurements arguably a regenerative medicine selection biomarker.
- Published
- 2020
40. Modeling Complex Neurological Diseases with Stem Cells: A Study of Bipolar Disorder
- Author
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Cameron D, Pernia, Neal H, Nathan, Brian T D, Tobe, Alicia M, Winquist, Richard L, Sidman, Yoshio, Goshima, and Evan Y, Snyder
- Subjects
Neurons ,Bipolar Disorder ,Induced Pluripotent Stem Cells ,Animals ,Humans ,Cell Differentiation ,Lithium - Abstract
The pathogenesis of bipolar disorder (BPD) is unknown. Using human-induced pluripotent stem cells (hiPSCs) to unravel pathological mechanisms in polygenic diseases is challenging, with few successful studies to date. However, hiPSCs from BPD patients responsive to lithium have offered unique opportunities to discern lithium's mechanism of action and hence gain insight into BPD pathology. By profiling the proteomics of BPD-hiPSC-derived neurons, we found that lithium alters the phosphorylation state of collapsin response mediator protein-2 (CRMP2). The "set point" for the ratio of pCRMP2:CRMP2 is elevated uniquely in hiPSC-derived neurons from lithium responsive (Li-R) BPD patients, but not other psychiatric and neurological disorders. Utilizing neurons differentiated from human patient stem cells as an in vitro platform, we were able to elucidate the mechanism driving the pathogenesis and pathophysiology of lithium-responsive BPD, heretofore unknown. Importantly, the findings in culture were validated in human postmortem material as well as in animal models of BPD behavior. These data suggest that the "lithium response pathway" in BPD governs CRMP2's phosphorylation, which regulates cytoskeletal organization, particularly in dendritic spines, leading to modulated neural networks that may underlie Li-R BPD pathogenesis. This chapter reviews the methodology of leveraging a functional agent, lithium, to identify unknown pathophysiological pathways with hiPSCs and how to translate this disease modeling approach to other neurological disorders.
- Published
- 2018
41. Modeling Complex Neurological Diseases with Stem Cells: A Study of Bipolar Disorder
- Author
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Brian T. D. Tobe, Alicia M. Winquist, Neal H. Nathan, Cameron D. Pernia, Evan Y. Snyder, Richard L. Sidman, and Yoshio Goshima
- Subjects
0301 basic medicine ,Dendritic spine ,business.industry ,Cellular differentiation ,Disease ,medicine.disease ,behavioral disciplines and activities ,Pathogenesis ,03 medical and health sciences ,030104 developmental biology ,mental disorders ,medicine ,Collapsin response mediator protein family ,Bipolar disorder ,Stem cell ,Induced pluripotent stem cell ,business ,Neuroscience - Abstract
The pathogenesis of bipolar disorder (BPD) is unknown. Using human-induced pluripotent stem cells (hiPSCs) to unravel pathological mechanisms in polygenic diseases is challenging, with few successful studies to date. However, hiPSCs from BPD patients responsive to lithium have offered unique opportunities to discern lithium’s mechanism of action and hence gain insight into BPD pathology. By profiling the proteomics of BPD–hiPSC-derived neurons, we found that lithium alters the phosphorylation state of collapsin response mediator protein-2 (CRMP2). The “set point” for the ratio of pCRMP2:CRMP2 is elevated uniquely in hiPSC-derived neurons from lithium responsive (Li-R) BPD patients, but not other psychiatric and neurological disorders. Utilizing neurons differentiated from human patient stem cells as an in vitro platform, we were able to elucidate the mechanism driving the pathogenesis and pathophysiology of lithium-responsive BPD, heretofore unknown. Importantly, the findings in culture were validated in human postmortem material as well as in animal models of BPD behavior. These data suggest that the “lithium response pathway” in BPD governs CRMP2’s phosphorylation, which regulates cytoskeletal organization, particularly in dendritic spines, leading to modulated neural networks that may underlie Li-R BPD pathogenesis. This chapter reviews the methodology of leveraging a functional agent, lithium, to identify unknown pathophysiological pathways with hiPSCs and how to translate this disease modeling approach to other neurological disorders.
- Published
- 2018
42. STEM-11. A MAPK-DRIVEN miR-124-SOX9 AXIS IS CRITICAL FOR STEM CELL MAINTENANCE, PROGRESSION, AND THERAPY-RESISTANCE IN GLIOBLASTOMA
- Author
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Ksenya Shchors, Selma Masic, Hanna Sabelström, Theo Nicolaides, Rebecca Petri, Evan Y. Snyder, Trenten Fenster, Shirin IIkhanizadeh, Edith Yuan, Johan Jakobsson, Rohit Sacheva, William A. Weiss, Rahul Jandial, Mitchel S. Berger, and Anders Persson
- Subjects
MAPK/ERK pathway ,Cancer Research ,business.industry ,SOX9 ,Biology ,medicine.disease ,nervous system diseases ,Abstracts ,Text mining ,Oncology ,medicine ,Cancer research ,Neurology (clinical) ,Treatment resistance ,Stem cell ,business ,Glioblastoma - Abstract
Glioblastoma (GBM) is the most common and aggressive malignant primary brain tumor. Genetic alterations in growth factor signaling pathways are found in 90% of GBMs. Advances in developmental and glioma biology suggest that common down-stream effector molecules in growth factor signaling pathways are critical for stem cell maintenance in the normal brain and GBM cells. It remains unclear whether differentiation therapies will be of therapeutic value for GBM patients. Here, we demonstrate that constitutive mitogen activated protein kinase (MAPK) activation in stem cells drives GBM formation and blocks neurogenesis in mice. Pharmacological inhibition of MAPK signaling restored neurogenesis in vivo and induced neuronal differentiation in GBM tumorspheres cultures established from murine GBMs and patient-derived tumors. Inhibition of MAPK signaling depleted SOX9 protein expression, and to a lesser extent SOX9 mRNA levels, in GBM cells. MicroRNA profiling experiments demonstrated that MAPK signaling regulates genome-wide expression of miRNAs, including the neuronal determinant miR-124. Pharmacological inhibition of MAPK signaling increased miR-124 levels in SOX9-expressing GBMs, but not SOX10-expressing proneural tumors. Using a doxycycline-inducible approach in vitro and in vivo, we demonstrated that miR-124 overexpression blocks SOX9 expression and induces neuronal differentiation in an EGFRvIII-driven GBM model and patient-derived xenografts. Neuronal differentiation resulted in apoptosis, reduced DNA repair capacity, and radiosensitized GBM cells. Doxycycline-mediated miR-124 overexpression resulted in complete regression for 1/3 of patient-derived xenografts. Mechanistic studies showed that SOX9 was a direct target of miR-124 and a major regulator of stem cell maintenance in GBM. Preliminary data showed that MAPK activation regulates transcriptional networks in SOX10-expressing proneural glioma, suggesting that distinct miRNAs regulate glioma aggressiveness in a subtype-specific manner. In conclusion, our results provide a mechanistic explanation for MAPK-dependent expansion of the stem cell pool during GBM initiation and demonstrate that enforcing neuronal differentiation represents a viable therapeutic strategy in glioma.
- Published
- 2017
43. hESC Differentiation toward an Autonomic Neuronal Cell Fate Depends on Distinct Cues from the Co-Patterning Vasculature
- Author
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Evan Y. Snyder, Breda M. Walsh, Flavio Cimadamore, Cameron D. Pernia, Lisette M. Acevedo, Alexey V. Terskikh, Martin S. Denzel, Connie Chen, Peik Sia, Jeffrey N. Lindquist, Barbara Ranscht, and David A. Cheresh
- Subjects
Male ,Cellular differentiation ,Human Embryonic Stem Cells ,Cell Culture Techniques ,Peripherins ,Ectoderm ,Biochemistry ,Muscle, Smooth, Vascular ,Mesoderm ,Mice ,0302 clinical medicine ,Tubulin ,Models ,Human embryogenesis ,RNA, Small Interfering ,lcsh:QH301-705.5 ,Cells, Cultured ,Mice, Knockout ,Neurons ,0303 health sciences ,lcsh:R5-920 ,Cultured ,Neural crest ,Cell Differentiation ,Cadherins ,Immunohistochemistry ,medicine.anatomical_structure ,Neural Crest ,Muscle ,RNA Interference ,Smooth ,lcsh:Medicine (General) ,Nitric Oxide Synthase Type III ,Cells ,Knockout ,Biology ,Cell fate determination ,Nitric Oxide ,Small Interfering ,Models, Biological ,Article ,03 medical and health sciences ,Vascular ,Genetics ,medicine ,Animals ,Humans ,Nitric Oxide Donors ,Cell Lineage ,Endothelium ,030304 developmental biology ,Neural tube ,Cell Biology ,Biological ,Embryonic stem cell ,Coculture Techniques ,lcsh:Biology (General) ,Epiblast ,Immunology ,Blood Vessels ,RNA ,Endothelium, Vascular ,Neuroscience ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Summary To gain insight into the cellular and molecular cues that promote neurovascular co-patterning at the earliest stages of human embryogenesis, we developed a human embryonic stem cell model to mimic the developing epiblast. Contact of ectoderm-derived neural cells with mesoderm-derived vasculature is initiated via the neural crest (NC), not the neural tube (NT). Neurovascular co-patterning then ensues with specification of NC toward an autonomic fate requiring vascular endothelial cell (EC)-secreted nitric oxide (NO) and direct contact with vascular smooth muscle cells (VSMCs) via T-cadherin-mediated homotypic interactions. Once a neurovascular template has been established, NT-derived central neurons then align themselves with the vasculature. Our findings reveal that, in early human development, the autonomic nervous system forms in response to distinct molecular cues from VSMCs and ECs, providing a model for how other developing lineages might coordinate their co-patterning., Graphical Abstract, Highlights • Neural crest (NC) cells drive neurovascular co-patterning, as modeled by hESC • Autonomic differentiation of NC cells depends on contact with perivascular cells • This requires endothelial-derived NO and T-cadherin-mediated interaction with VSMCs, Cheresh, Snyder, and colleagues demonstrate how direct contact between hESC-derived neural crest (NC) cells and the vasculature promotes autonomic differentiation. In a hESC model mimicking the developing epiblast, NC cells initiate neurovascular co-patterning prior to forming neural tube neurons. Endothelial-secreted NO and direct contact with vascular smooth muscle cells via T-cadherin are required to promote NC differentiation into autonomic neurons.
- Published
- 2015
44. Enabling Consistency in Pluripotent Stem Cell-Derived Products for Research and Development and Clinical Applications Through Material Standards
- Author
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Amit Chandra, Mahendra S. Rao, Hae-Won Kim, Doug Sipp, Glyn Stacey, Guillermo García-Cardeña, Joseph C. Wu, Alexander Meissner, Jonathan C. Knowles, Natalie Artzi, Joseph D. Gold, Andrew Carr, David Brindley, Ivan B. Wall, Christopher A. Bravery, David J. Williams, Peter R.T. Archibald, Richard W. Barker, James R. Smith, Kim Bure, Anna French, Jeanne F. Loring, Brock Reeve, Evan Y. Snyder, Jeffrey M. Karp, and Steve Oh
- Subjects
Pluripotent Stem Cells ,education.field_of_study ,Biomedical Research ,business.industry ,Population ,Drug Evaluation, Preclinical ,Cell Biology ,General Medicine ,Reference Standards ,Consistency (database systems) ,Risk analysis (engineering) ,Basic research ,Physical form ,Humans ,Medicine ,Clinical safety ,business ,education ,Induced pluripotent stem cell ,Reference standards ,Perspectives ,Developmental Biology - Abstract
Summary There is a need for physical standards (reference materials) to ensure both reproducibility and consistency in the production of somatic cell types from human pluripotent stem cell (hPSC) sources. We have outlined the need for reference materials (RMs) in relation to the unique properties and concerns surrounding hPSC-derived products and suggest in-house approaches to RM generation relevant to basic research, drug screening, and therapeutic applications. hPSCs have an unparalleled potential as a source of somatic cells for drug screening, disease modeling, and therapeutic application. Undefined variation and product variability after differentiation to the lineage or cell type of interest impede efficient translation and can obscure the evaluation of clinical safety and efficacy. Moreover, in the absence of a consistent population, data generated from in vitro studies could be unreliable and irreproducible. Efforts to devise approaches and tools that facilitate improved consistency of hPSC-derived products, both as development tools and therapeutic products, will aid translation. Standards exist in both written and physical form; however, because many unknown factors persist in the field, premature written standards could inhibit rather than promote innovation and translation. We focused on the derivation of physical standard RMs. We outline the need for RMs and assess the approaches to in-house RM generation for hPSC-derived products, a critical tool for the analysis and control of product variation that can be applied by researchers and developers. We then explore potential routes for the generation of RMs, including both cellular and noncellular materials and novel methods that might provide valuable tools to measure and account for variation. Multiparametric techniques to identify “signatures” for therapeutically relevant cell types, such as neurons and cardiomyocytes that can be derived from hPSCs, would be of significant utility, although physical RMs will be required for clinical purposes.
- Published
- 2015
45. Generating iPSCs: Translating Cell Reprogramming Science into Scalable and Robust Biomanufacturing Strategies
- Author
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Andrew Carr, Brock Reeve, Laurence Daheron, Anna French, Hae-Won Kim, Kelvin S. Ng, Kim Bure, Peter J. Coffey, Richard J. Barker, David J. Williams, Mahendra Rao, Hannah Hurley, Jeffrey M. Karp, James A. Smith, David Brindley, Benjamin E. Mead, Ivan Wall, Marli Silva, Evan Y. Snyder, and Justin J. Cooper-White
- Subjects
Drug discovery ,Induced Pluripotent Stem Cells ,Cell Culture Techniques ,Reproducibility of Results ,Cell Biology ,Biology ,Cellular Reprogramming ,Bioinformatics ,Data science ,Scalability ,Genetics ,Animals ,Humans ,Molecular Medicine ,Biomanufacturing ,Induced pluripotent stem cell ,Reprogramming ,Stem Cell Transplantation - Abstract
Induced pluripotent stem cells (iPSCs) have the potential to transform drug discovery and healthcare in the 21(st) century. However, successful commercialization will require standardized manufacturing platforms. Here we highlight the need to define standardized practices for iPSC generation and processing and discuss current challenges to the robust manufacture of iPSC products.
- Published
- 2015
46. The state of the art in stem cell biology and regenerative medicine: the end of the beginning
- Author
-
Evan Y. Snyder
- Subjects
0301 basic medicine ,Cellular basis ,Proteomics ,Regenerative Medicine ,Regenerative medicine ,Quarter century ,Translational Research, Biomedical ,03 medical and health sciences ,Mice ,Neural Stem Cells ,Animals ,Humans ,Cell Lineage ,Precision Medicine ,Child ,Organism ,Cognitive science ,Gene Editing ,Neuronal Plasticity ,Pediatric research ,Stem Cells ,030104 developmental biology ,Pediatrics, Perinatology and Child Health ,Stem cell ,Developmental biology ,Stem cell biology ,Developmental Biology ,Stem Cell Transplantation - Abstract
With translational stem cell biology and Regenerative Medicine (the field to which the former gave rise) now over a quarter century old, it is time to take stock of where we have been and where we are going. This editorial overview, which serves as an introduction to this special issue of Pediatric Research dedicated to these fields, reinforces the notion that stem cells are ultimately intrinsic parts of developmental biology, for which Pediatrics represents the clinical face. Although stem cells provide the cellular basis for a great deal of only recently recognized plasticity programmed into the developing and postdevelopmental organism, and although there is enormous promise in harnessing this plasticity for therapeutic advantage, their successful use rests on a deep understanding of their developmental imperatives and the developmental programs in which they engage. The potential uses of stems are ranked and discussed in the order of most readily achievable to those requiring extensively more work. Although that order may not be what was contemplated at the field’s birth, we nevertheless retain an optimism for the ultimate positive impact of exploiting this fundamental biology for the well-being of children.
- Published
- 2017
47. Stem Cell Technologies in Neuroscience
- Author
-
Yang D. Teng, Evan Y. Snyder, and Amit K. Srivastava
- Subjects
Chemistry ,Stem cell ,Neuroscience - Published
- 2017
48. Multimodal Neural Stem Cell Research Protocols for Experimental Spinal Cord Injuries
- Author
-
Liquan Wu, Evan Y. Snyder, Xiang Zeng, Inbo Han, and Yang D. Teng
- Subjects
medicine.anatomical_structure ,medicine ,Biology ,Spinal cord ,Neuroscience ,Neural stem cell - Published
- 2017
49. Probing the lithium-response pathway in hiPSCs implicates the phosphoregulatory set-point for a cytoskeletal modulator in bipolar pathogenesis
- Author
-
Fumio Nakamura, Husseini K. Manji, Namrata D. Udeshi, Hagop S. Akiskal, Jeffrey H. Price, Nikolaos Venizelos, Wen-Ning Zhao, Cordulla Duerr, Ryan W. Logan, Jeffrey S. Nye, Jasmin Lalonde, Steven D. Sheridan, Joseph T. Coyle, Toshio Ohshima, Yoshio Goshima, Michael McCarthy, Shelley Halpain, Barbara Calabrese, Joshua G. Hunsberger, Martin Alda, Yuuka Inoue, Gustavo J. Gutierrez, Stephen J. Haggarty, Brian T. D. Tobe, Cameron D. Pernia, Richard L. Sidman, Moyuka Wada, Hiroko Makihara, Laurence M. Brill, Glenn T. Konopaske, Yang Liu, De-Maw M. Chuang, Ranor C. B. Basa, Alicia M. Winquist, Yang D. Teng, Michelle M. Sidor, Steven A. Carr, Colleen A. McClung, Guy A. Rouleau, Katsuhiko Mikoshiba, Evan Y. Snyder, Laurel Dorsett, Lina Mastrangelo, Guang Chen, Jianxue Li, Haruko Nakamura, Andrew Crain, Philipp Mertins, Michael G. Brandel, Dongmei Wu, Naoya Yamashita, and Biology
- Subjects
0301 basic medicine ,Genetically modified mouse ,Proteomics ,Dendritic spine ,Bipolar Disorder ,Induced Pluripotent Stem Cells ,Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) ,Nerve Tissue Proteins ,Biology ,Lithium ,Models, Biological ,Psykiatri ,03 medical and health sciences ,Mice ,0302 clinical medicine ,posttranslational modification ,proteomics ,psychiatric disease modeling ,CRMP2 ,dendrites ,Calcium flux ,mental disorders ,medicine ,Animals ,Humans ,Induced pluripotent stem cell ,Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci) ,Cells, Cultured ,Brain Chemistry ,Psychiatry ,Multidisciplinary ,Neurosciences ,030104 developmental biology ,medicine.anatomical_structure ,PNAS Plus ,Phosphorylation ,Intercellular Signaling Peptides and Proteins ,Calcium ,Neuron ,Collapsin response mediator protein family ,Neuroscience ,Protein Processing, Post-Translational ,030217 neurology & neurosurgery ,Intracellular ,Neurovetenskaper - Abstract
The molecular pathogenesis of bipolar disorder (BPD) is poorly understood. Using human-induced pluripotent stem cells (hiPSCs) to unravel such mechanisms in polygenic diseases is generally challenging. However, hiPSCs from BPD patients responsive to lithium offered unique opportunities to discern lithium's target and hence gain molecular insight into BPD. By profiling the proteomics of BDP-hiPSC-derived neurons, we found that lithium alters the phosphorylation state of collapsin response mediator protein-2 (CRMP2). Active non-phosphorylated CRMP2, which binds cytoskeleton, is present throughout the neuron; inactive phosphorylated CRMP2, which dissociates from cytoskeleton, exits dendritic spines. CRMP2 elimination yields aberrant dendritogenesis with diminished spine density and lost lithium responsiveness (LiR). The "set-point" for the ratio of pCRMP2: CRMP2 is elevated uniquely in hiPSC-derived neurons from LiR BPD patients, but not with other psychiatric (including lithium-nonresponsive BPD) and neurological disorders. Lithium (and other pathway modulators) lowers pCRMP2, increasing spine area and density. Human BPD brains show similarly elevated ratios and diminished spine densities; lithium therapy normalizes the ratios and spines. Consistent with such "spine-opathies," human LiR BPD neurons with abnormal ratios evince abnormally steep slopes for calcium flux; lithium normalizes both. Behaviorally, transgenic mice that reproduce lithium's postulated site-of-action in dephosphorylating CRMP2 emulate LiR in BPD. These data suggest that the " lithium response pathway" in BPD governs CRMP2's phosphorylation, which regulates cytoskeletal organization, particularly in spines, modulating neural networks. Aberrations in the posttranslational regulation of this developmentally critical molecule may underlie LiR BPD pathogenesis. Instructively, examining the proteomic profile in hiPSCs of a functional agent-even one whose mechanism-of-action is unknown-might reveal otherwise inscrutable intracellular pathogenic pathways., Funding Agencies:NIH's Library of Integrated Network-based Cellular Signatures Program Viterbi Foundation Neuroscience Initiative Stanley Medical Research Institute R21MH093958 R33MH087896 R01MH095088 Tau Consortium California Institute of Regenerative Medicine training grants University of California, San Diego T32 training grant in psychiatry California Bipolar Foundation International Bipolar Foundation Creation of Innovation Centers for Advanced Interdisciplinary Research Areas Program in the Project for Developing Innovation Systems from the Ministry of Education, Science, Sports and Culture in Japan 42890001 RC2MH090011
- Published
- 2017
50. Contributors
- Author
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Gregory S. Aaen, Nicholas Scott Abend, Amal Abou-Hamden, Jeffrey C. Allen, Anthony A. Amato, Catherine Amlie-Lefond, Stephen Ashwal, Russell C. Bailey, James F. Bale, Brenda Banwell, Kristin W. Barañano, A. James Barkovich, Richard J. Barohn, Ute K. Bartels, Brenda Bartnik-Olson, Ori Barzilai, Alexander Bassuk, David R. Bearden, Liat Ben-Sira, Timothy J. Bernard, Elizabeth Berry-Kravis, Lauren A. Beslow, Jaclyn A. Biegel, Lori Billinghurst, Angela K. Birnbaum, Joanna S. Blackburn, Nuala Bobowski, Adrienne Boire, Carsten G. Bönnemann, Sonia L. Bonifacio, Daniel J. Bonthius, Breck Borcherding, Brian R. Branchford, John Brandsema, Kathryn M. Brennan, J. Nicholas Brenton, Amy R. Brooks-Kayal, Lawrence W. Brown, Jeffrey Buchalter, Carol S. Camfield, Peter R. Camfield, Cristina Campoy, Jessica L. Carpenter, Taeun Chang, Vann Chau, Susan N. Chi, Claudia A. Chiriboga, Yoon-Jae Cho, Cindy W. Christian, Nicholas Chrestian, Maria Roberta Cilio, Robin D. Clark, Bruce H. Cohen, Ronald D. Cohn, Anne M. Connolly, Todd Constable, Shlomi Constantini, Jeannine M. Conway, David L. Coulter, Tina M. Cowan, Russell C. Dale, Benjamin Darbro, Basil T. Darras, Jahannaz Dastgir, Linda De Meirleir, Darryl C. De Vivo, Linda S. de Vries, Jeremy K. Deisch, Paul Deltenre, Jay Desai, Maria Descartes, Gabrielle deVeber, Sameer C. Dhamne, Jullianne Diaz, Salvatore DiMauro, William B. Dobyns, Dan Doherty, Elizabeth J. Donner, Nico U.F. Dosenbach, James J. Dowling, James M. Drake, Cecile Ejerskov, Andrew G. Engel, Gregory M. Enns, María Victoria Escolano-Margarit, Iris Etzion, S. Ali Fatemi, Darcy L. Fehlings, Michelle Lauren Feinberg, Donna M. Ferriero, Pauline A. Filipek, Richard S. Finkel, Paul G. Fisher, Kevin Flanigan, Nicholas K. Foreman, Israel Franco, Yitzchak Frank, Douglas R. Fredrick, Hudson H. Freeze, Cristina Fuente-Mora, Joseph M. Furman, Renata C. Gallagher, Catherine Garel, Emily Gertsch, Donald L. Gilbert, Elizabeth E. Gilles, Christopher C. Giza, Carol A. Glaser, Hannah C. Glass, Tracy Glauser, Joseph Glykys, Amy Goldstein, Hernan Dario Gonorazky, Rodolfo Gonzalez, Howard P. Goodkin, John M. Graham, Alexander L. Greninger, Gary Gronseth, Andrea L. Gropman, Richard Grundy, Renzo Guerrini, Nalin Gupta, Jin S. Hahn, Milton H. Hamblin, Abeer J. Hani, Sharyu Hanmantgad, Mary J. Harbert, Chellamani Harini, Andrea M. Harriott, Chad Heatwole, Andrew D. Hershey, Deborah G. Hirtz, Gregory L. Holmes, Barbara A. Holshouser, Kathleen A. Hurwitz, Eugene Hwang, Rebecca N. Ichord, Paymaan Jafar-Nejad, Sejal V. Jain, Lori Jordan, Marielle A. Kabbouche, Joanne Kacperski, Peter B. Kang, Matthias A. Kariannis, Horacio Kaufmann, Harper L. Kaye, Robert Keating, Colin R. Kennedy, Yasmin Khakoo, Adam Kirton, John T. Kissel, Kelly G. Knupp, Bruce R. Korf, Eric H. Kossoff, Sanjeev V. Kothare, Oren Kupfer, W. Curt LaFrance, Beatrice Latal, Steven M. Leber, Jean-Pyo Lee, Ilo E. Leppik, Tally Lerman-Sagie, Jason T. Lerner, Richard J. Leventer, Daniel J. Licht, Uta Lichter-Konecki, Zvi Lidar, Djin Gie Liem, Tobias Loddenkemper, Roger K. Long, Quyen N. Luc, Mark Mackay, Annette Majnemer, Naila Makhani, Gustavo Malinger, David E. Mandelbaum, Stephen M. Maricich, Kiran P. Maski, Mudit Mathur, Dennis J. Matthews, Kelly McMahon, Megan B. DeMara-Hoth, Bryce Mendelsohn, Julie A. Mennella, Laura R. Ment, Eugenio Mercuri, David J. Michelson, Mohamad A. Mikati, Fady M. Mikhail, Steven Paul Miller, Jeff M. Milunsky, Jonathan W. Mink, Ghayda M. Mirzaa, Wendy G. Mitchell, Michael A. Mohan, Payam Mohassel, Mahendranath Moharir, Umrao R. Monani, Michelle Monje Deisseroth, Manikum Moodley, Andrew Mower, Richard T. Moxley, Sabine Mueller, Alysson R. Muotri, Sandesh C.S. Nagamani, Mohan J. Narayanan, Vinodh Narayanan, Ruth D. Nass, Jeffrey L. Neul, Yoram Nevo, Bobby G. Ng, Katherine C. Nickels, Graeme A.M. Nimmo, Michael J. Noetzel, Lucy Norcliffe-Kaufmann, Douglas R. Nordli, Ulrike Nowak-Göttl, Hope L. O'Brien, Joyce Oleszek, Maryam Oskoui, Alex R. Paciorkowski, Roger J. Packer, Seymour Packman, Jose-Alberto Palma, Andrea C. Pardo, Julie A. Parsons, John Colin Partridge, Gregory M. Pastores, Marc C. Patterson, William J. Pearce, Phillip L. Pearl, Melanie Penner, Leila Percival, Marcia Pereira, Stefan M. Pfister, John Phillips, Barbara Plecko, Sigita Plioplys, Annapurna Poduri, Sharon Poisson, Scott L. Pomeroy, Andrea Poretti, Scott W. Powers, Michael R. Pranzatelli, Allison Przekop, Malcolm Rabie, Sampathkumar Rangasamy, Gerald V. Raymond, Alyssa T. Reddy, Rebecca L. Rendleman, Jong M. Rho, Lance H. Rodan, Sarah M. Roddy, Elizabeth E. Rogers, Stephen M. Rosenthal, N. Paul Rosman, M. Elizabeth Ross, Alexander Rotenberg, Robert S. Rust, Cheryl P. Sanchez, Pedro Sanchez, Iván Sánchez Fernández, Tristan T. Sands, Terence D. Sanger, Kumar Sannagowdara, Dustin Scheinost, Mark S. Scher, Nina F. Schor, Isabelle Schrauwen, Michael M. Segal, Syndi Seinfeld, Duygu Selcen, Laurie E. Seltzer, Margaret Semrud-Clikeman, Dennis W. Shaw, Bennett A. Shaywitz, Sally E. Shaywitz, Renée A. Shellhaas, Elliott H. Sherr, Rita D. Sheth, Michael I. Shevell, Shlomo Shinnar, Ben Shofty, Stanford K. Shu, Michael E. Shy, Laura Silveira Moriyama, Nicholas J. Silvestri, Katherine B. Sims, Harvey S. Singer, Nilika Shah Singhal, Craig M. Smith, Edward Smith, Stephen A. Smith, Evan Y. Snyder, Janet Soul, Christy L. Spalink, Karen A. Spencer, Carl E. Stafstrom, Robert Steinfeld, Jonathan B. Strober, Joseph Sullivan, Kenneth F. Swaiman, Kathryn J. Swoboda, Elizabeth D. Tate, William O. Tatum, Ingrid Tein, Kristyn Tekulve, Jeffrey R. Tenney, Elizabeth A. Thiele, Robert Thompson-Stone, Laura Tochen, Laura M. Tormoehlen, Lily Tran, Doris A. Trauner, Sinan O. Turnacioglu, Nicole J. Ullrich, David K. Urion, Guy Van Camp, Michèle Van Hirtum-Das, Clara D.M. van Karnebeek, Lionel Van Maldergem, Adeline Vanderver, Nicholas A. Vitanza, Michael von Rhein, Emily von Scheven, Ann Wagner, Mark S. Wainwright, Melissa A. Walker, John T. Walkup, Laurence Walsh, Lauren C. Walters-Sen, Raymond Y. Wang, Thomas T. Warner, Harry T. Whelan, Geoffrey A. Weinberg, Elizabeth M. Wells, James W. Wheless, Elaine C. Wirrell, Jeffrey H. Wisoff, Nicole I. Wolf, Gil I. Wolfe, F. Virginia Wright, Nathaniel D. Wycliffe, Michele L. Yang, Christopher J. Yuskaitis, Huda Y. Zoghbi, and Mary L. Zupanc
- Published
- 2017
Catalog
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