• transdifferentiation
• direct conversion
• small molecule
• chemical cocktail
• neural cells

• Cell Transdifferentiation/drug effects
• Humans
• Neurons/cytology
• Neurons/physiology
• Neurons/drug effects
• Regenerative Medicine/methods

• Epigenetics
• IGF2BPs
• Reprogramming
• Stem cells
• iPSC
• m6A

• Humans
• Animals
• Mice
• Induced Pluripotent Stem Cells/metabolism
• Cell Differentiation/genetics
• Epigenesis, Genetic
• Fibroblasts/metabolism
• Cellular Reprogramming/genetics

• BX002905 VA
• California Institute of Regenerative Medicine (CIRM) grant
• Department of Veterans Affairs
• Key Technologies Research and Development Program
• the Innovative Program of National Natural Science Foundation of China
• the National Natural Science Foundation of China
• Fund of Jilin Provincial Finance Department
• Fund of Jilin Provincial Development and Reform Commission
• Fund of Jilin Provincial Health Commission

• Animals
• Brain
• Humans
• Lab-On-A-Chip Devices
• Microfluidics
• Neurons
• Tissue Engineering

• Joachim Herz Stiftung
• Paul-Ehrlich-Gesellschaft Stiftung
• Volkswagen Foundation
• Deutsche Forschungsgemeinschaft
• Pro Retina-Stiftung

• Human induced pluripotent stem cells
• Reprogramming
• Self-renewal
• Small molecules

• Cellular Reprogramming/genetics
• Humans
• Induced Pluripotent Stem Cells
• Phosphorylation
• Protein Kinases
• Protein Serine-Threonine Kinases/antagonists & inhibitors
• Protein-Tyrosine Kinases/antagonists & inhibitors
• Pyruvate Dehydrogenase Acetyl-Transferring Kinase/metabolism
• Dyrk Kinases

• ROCK
• Schlemm’s canal
• glaucoma
• intraocular pressure
• rho kinase inhibitors
• trabecular meshwork

• P30 EY020799 NEI NIH HHS

Neural crest cells (NCCs) are a promising source for cell therapy and regenerative medicine owing to their multipotency, self-renewability, and capability to secrete various trophic factors. However, isolating NCCs from adult organs is challenging, because NCCs are broadly distributed throughout the body. Hence, we attempted to directly induce NCCs from human adipose-derived mesenchymal stem cells (ADSCs), which can be isolated easily, using small molecule cocktails. We established a controlled induction protocol with two-step application of small molecule cocktails for 6 days. The induction efficiency was evaluated based on mRNA and protein expression of neural crest markers, such as nerve growth factor receptor (NGFR) and sex-determining region Y-box 10 (SOX10). We also found that various trophic factors were significantly upregulated following treatment with the small molecule cocktails. Therefore, we performed global profiling of cell surface makers and identified distinctly upregulated markers, including the neural crest-specific cell surface markers CD271 and CD57. These results indicate that our chemical treatment can direct human ADSCs to developing into the neural crest lineage. This offers a promising experimental platform to study human NCCs for applications in cell therapy and regenerative medicine.

• CD57 Antigens/metabolism
• Cell Culture Techniques
• Cell Differentiation
• Cell Proliferation
• Cells, Cultured
• Culture Media
• Humans
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Nerve Tissue Proteins/metabolism
• Neural Crest/cytology
• Neural Crest/metabolism
• Receptors, Nerve Growth Factor/metabolism
• Regenerative Medicine/methods
• SOXE Transcription Factors/metabolism

• Japan Society for the Promotion of Science
• Biomimetics Sympathies Inc.

• approaches
• epigenetic
• mechanisms
• molecular docking
• reprogramming
• small molecules

• Beta cells
• Differentiation
• Mesenchymal stem cells
• Small molecules
• Therapy
• Transplantation

Human mesenchymal stem cells (MSCs) are good candidates for brain cell replacement strategies and have already been used as adjuvant treatments in neurological disorders. MSCs can be obtained from many different sources, and the present study compares the potential of neuronal transdifferentiation in MSCs from adult and neonatal sources (Wharton's jelly (WhJ), dental pulp (DP), periodontal ligament (PDL), gingival tissue (GT), dermis (SK), placenta (PLAC), and umbilical cord blood (UCB)) with a protocol previously tested in bone marrow- (BM-) MSCs consisting of a cocktail of six small molecules: I-BET151, CHIR99021, forskolin, RepSox, Y-27632, and dbcAMP (ICFRYA). Neuronal morphology and the presence of cells positive for neuronal markers (TUJ1 and MAP2) were considered attributes of neuronal induction. The ICFRYA cocktail did not induce neuronal features in WhJ-MSCs, and these features were only partial in the MSCs from dental tissues, SK-MSCs, and PLAC-MSCs. The best response was found in UCB-MSCs, which was comparable to the response of BM-MSCs. The addition of neurotrophic factors to the ICFRYA cocktail significantly increased the number of cells with complex neuron-like morphology and increased the number of cells positive for mature neuronal markers in BM- and UCB-MSCs. The neuronal cells generated from UCB-MSCs and BM-MSCs showed increased reactivity of the neuronal genes TUJ1, MAP2, NF-H, NCAM, ND1, TAU, ENO2, GABA, and NeuN as well as down- and upregulation of MSC and neuronal genes, respectively. The present study showed marked differences between the MSCs from different sources in response to the transdifferentiation protocol used here. These results may contribute to identifying the best source of MSCs for potential cell replacement therapies.

• Arthritis
• degenerative disease
• diabetes
• endogenous stem cells
• heart disease
• muscular dystrophy
• regenerative medicine
• small molecules
• stem cell niche
• tissue repair

The combination of defined factors with small molecules targeting epigenetic factors is a strategy that has been shown to enhance optimal derivation of iPSCs and could be used for disease modelling, high throughput screenings and/or regenerative medicine applications. In this study, we showed that a new first-in-class reversible dual G9a/DNMT1 inhibitor compound (CM272) improves the efficiency of human cell reprogramming and iPSC generation from primary cells of healthy donors and patient samples, using both integrative and non-integrative methods. Moreover, CM272 facilitates the generation of human iPSC with only two factors allowing the removal of the most potent oncogenic factor cMYC. Furthermore, we demonstrated that mechanistically, treatment with CM272 induces heterochromatin relaxation, facilitates the engagement of OCT4 and SOX2 transcription factors to OSKM refractory binding regions that are required for iPSC establishment, and enhances mesenchymal to epithelial transition during the early phase of cell reprogramming. Thus, the use of this new G9a/DNMT reversible dual inhibitor compound may represent an interesting alternative for improving cell reprogramming and human iPSC derivation for many different applications while providing interesting insights into reprogramming mechanisms.

• Cells, Cultured
• Cellular Reprogramming
• Genome, Human
• Histocompatibility Antigens
• Histone-Lysine N-Methyltransferase/antagonists & inhibitors
• Humans
• Induced Pluripotent Stem Cells/cytology
• Real-Time Polymerase Chain Reaction
• Repressor Proteins/antagonists & inhibitors
• Transcription Factors/metabolism

• Instituto de Salud Carlos III
• CIBERONC. Instituto de Salud Carlos III
• TERCEL. Instituto de Salud Carlos III (ES).
• ERA-NET TRANSCAN-2. European Commission. Horizon2020
• ERA-NET EuroNanoMed-II. European Commission. FP7
• Secretaría de Estado de Investigación, Desarrollo e Innovación
• Fundación Caja Navarra

Brown adipocytes play an important role in human energy metabolism and prevention of obesity and diabetes. Induced pluripotent stem cells (iPSCs) represent a promising source for brown adipocytes; however, exogenous gene induction is generally required for iPSCs generation, which might cause undesired effects particularly in long-term treatment after transplantation. We have previously reported a cocktail of six small chemical compounds that enables a conversion of human fibroblasts into chemical compound-induced neuronal cells (CiNCs). Here, we report that modified combinations of the chemical compounds and rosiglitazone, a PPARγ agonist, afforded direct conversion of human fibroblasts into brown adipocytes. The chemical compound-induced brown adipocytes (ciBAs) exhibit induction of human brown adipocyte-specific genes such as Ucp1, Ckmt1, Cited1 and other adipocyte-specific genes such as Fabp4, AdipoQ, and Pparγ. Treatment with either isoproterenol or Forskolin further induced the expression of Ucp1, suggesting that β adrenergic receptor signalling in ciBAs could be functional for induction of thermogenic genes. Moreover, oxygen consumption rates were elevated in ciBAs along with increase of cellular mitochondria. Our findings might provide an easily accessible approach for generating human brown adipocytes from fibroblasts and offer therapeutic potential for the management of obesity, diabetes, and related metabolic disorders.

• Developmental reprogramming
• Physcomitrella patens
• Protoplast
• Quantitative proteome

• Bryopsida/metabolism
• Plant Proteins/metabolism
• Proteomics/methods
• Protoplasts/metabolism
• Signal Transduction/physiology

The transport of drug molecules is mainly determined by the distribution of influx and efflux transporters for which they are substrates. To enable tissue targeting, we sought to develop the idea that we might affect the transporter-mediated disposition of small-molecule drugs via the addition of a second small molecule that of itself had no inhibitory pharmacological effect but that influenced the expression of transporters for the primary drug. We refer to this as a “binary weapon” strategy. The experimental system tested the ability of a molecule that on its own had no cytotoxic effect to increase the toxicity of the nucleoside analog gemcitabine to Panc1 pancreatic cancer cells. An initial phenotypic screen of a 500-member polar drug (fragment) library yielded three “hits.” The structures of 20 of the other 2,000 members of this library suite had a Tanimoto similarity greater than 0.7 to those of the initial hits, and each was itself a hit (the cheminformatics thus providing for a massive enrichment). We chose the top six representatives for further study. They fell into three clusters whose members bore reasonable structural similarities to each other (two were in fact isomers), lending strength to the self-consistency of both our conceptual and experimental strategies. Existing literature had suggested that indole-3-carbinol might play a similar role to that of our fragments, but in our hands it was without effect; nor was it structurally similar to any of our hits. As there was no evidence that the fragments could affect toxicity directly, we looked for effects on transporter transcript levels. In our hands, only the ENT1-3 uptake and ABCC2,3,4,5, and 10 efflux transporters displayed measurable transcripts in Panc1 cultures, along with a ribonucleoside reductase RRM1 known to affect gemcitabine toxicity. Very strikingly, the addition of gemcitabine alone increased the expression of the transcript for ABCC2 (MRP2) by more than 12-fold, and that of RRM1 by more than fourfold, and each of the fragment “hits” served to reverse this. However, an inhibitor of ABCC2 was without significant effect, implying that RRM1 was possibly the more significant player. These effects were somewhat selective for Panc cells. It seems, therefore, that while the effects we measured were here mediated more by efflux than influx transporters, and potentially by other means, the binary weapon idea is hereby fully confirmed: it is indeed possible to find molecules that manipulate the expression of transporters that are involved in the bioactivity of a pharmaceutical drug. This opens up an entirely new area, that of chemical genomics-based drug targeting.

• anticancer drugs
• binary weapon
• cheminformatics
• drug transporters
• gemcitabine
• pancreatic cancer
• phenotypic screening

Forced expression of lineage-specific transcription factors in somatic cells can result in the generation of different cell types in a process named direct reprogramming, bypassing the pluripotent state. However, the introduction of transgenes limits the therapeutic applications of the produced cells. Numerous small-molecules have been introduced in the field of stem cell biology capable of governing self-renewal, reprogramming, transdifferentiation and regeneration. These chemical compounds are versatile tools for cell fate conversion toward desired outcomes. Cell fate conversion using small-molecules alone (chemical reprogramming) has superiority over arduous traditional genetic techniques in several aspects. For instance, rapid, transient, and reversible effects in activation and inhibition of functions of specific proteins are of the profits of small-molecules. They are cost-effective, have a long half-life, diversity on structure and function, and allow for temporal and flexible regulation of signaling pathways. Additionally, their effects could be adjusted by fine-tuning concentrations and combinations of different small-molecules. Therefore, chemicals are powerful tools in cell fate conversion and study of stem cell and chemical biology in vitro and in vivo. Moreover, transgene-free and chemical-only transdifferentiation approaches provide alternative strategies for the generation of various cell types, disease modeling, drug screening, and regenerative medicine. The current review gives an overview of the recent findings concerning transdifferentiation by only small-molecules without the use of transgenes.

• Cellular reprogramming
• Regenerative medicine
• Small-molecule
• Transdifferentiation

• Akt
• FoxO
• SIRT1
• WISP1
• Wnt signaling
• apoptosis
• autophagy
• cell longevity
• erythropoietin
• forkhead transcription factors
• growth factors
• mechanistic target of rapamycin (mTOR)
• mitochondria
• oxidative stress
• programmed cell death
• reactive oxygen species
• sirtuins
• stem cells
• transcription factors

• R01 NS053946 NINDS NIH HHS

Stem cells offer great promise for the treatment of multiple disorders throughout the body. Critical to this premise is the ability to govern stem cell pluripotency, proliferation, and differentiation. The mechanistic target of rapamycin (mTOR), 289-kDa serine/threonine protein kinase, that is a vital component of mTOR Complex 1 and mTOR Complex 2 represents a critical pathway for the oversight of stem cell maintenance. mTOR can control the programmed cell death pathways of autophagy and apoptosis that can yield variable outcomes in stem cell survival and be reliant upon proliferative pathways that include Wnt signaling, Wnt1 inducible signaling pathway protein 1 (WISP1), silent mating type information regulation 2 homolog 1 (Saccharomyces cerevisiae) (SIRT1), and trophic factors. mTOR also is a necessary component for the early development and establishment of stem cells as well as having a significant impact in the regulation of the maturation of specific cell phenotypes. Yet, as a proliferative agent, mTOR can not only foster cancer stem cell development and tumorigenesis, but also mediate cell senescence under certain conditions to limit invasive cancer growth. mTOR offers an exciting target for the oversight of stem cell therapies but requires careful consideration of the diverse clinical outcomes that can be fueled by mTOR signaling pathways.

• Apoptosis
• Autophagy
• Cancer
• Cardiovascular
• Erythropoietin
• Mechanistic target of rapamycin
• Neurodegeneration
• Progenitor stem cells
• Silent mating type information regulation 2 homolog
• Wnt1 inducible signaling pathway
• Wnt

• Animals
• Epigenesis, Genetic
• Forkhead Transcription Factors/genetics
• Forkhead Transcription Factors/metabolism
• Humans
• Nervous System/metabolism
• Nervous System Diseases/metabolism
• Nervous System Diseases/pathology
• Oxidative Stress
• Protein Processing, Post-Translational

• ipscs
• mitochondria
• metabolism
• stem cells
• reprogramming
• pluripotency

• Animals
• Cell Proliferation
• Cellular Reprogramming
• Energy Metabolism
• Humans
• Metabolic Networks and Pathways
• Mitochondria/genetics
• Mitochondria/metabolism
• Pluripotent Stem Cells/cytology
• Pluripotent Stem Cells/metabolism

• AMPK
• Alzheimer's disease
• CCN4
• EGF
• EPO
• FGF
• IGF-1
• WISP1
• Wnt
• apoptosis
• autophagy
• central nervous system
• diabetes mellitus
• erythropoietin
• mTOR
• neuron
• neuropathy
• oxidative stress
• psychiatric
• stem cells

Induced pluripotent stem cells (iPSCs) could be employed in the creation of patient-specific stem cells, which could subsequently be used in various basic and clinical applications. However, current iPSC methodologies present significant hidden risks with respect to genetic mutations and abnormal expression which are a barrier in realizing the full potential of iPSCs. A chemical approach is thought to be a promising strategy for safety and efficiency of iPSC generation. Many small molecules have been identified that can be used in place of exogenous transcription factors and significantly improve iPSC reprogramming efficiency and quality. Recent studies have shown that the use of small molecules results in the generation of chemically induced pluripotent stem cells from mouse embryonic fibroblast cells. These studies might lead to new areas of stem cell research and medical applications, not only human iPSC by chemicals alone, but also safe generation of somatic stem cells for cell based clinical trials and other researches. In this paper, we have reviewed the recent advances in small molecule approaches for the generation of iPSCs.

Pluripotent stem cells are unspecialized cells with unlimited self-renewal, and they can be triggered to differentiate into desired specialized cell types. These features provide the basis for an unlimited cell source for innovative cell therapies. Pluripotent cells also allow to study developmental pathways, and to employ them or their differentiated cell derivatives in pharmaceutical testing and biotechnological applications. Via blastocyst complementation, pluripotent cells are a favoured tool for the generation of genetically modified mice. The recently established technology to generate an induced pluripotency status by ectopic co-expression of the transcription factors Oct4, Sox2, Klf4 and c-Myc allows to extending these applications to farm animal species, for which the derivation of genuine embryonic stem cells was not successful so far. Most induced pluripotent stem (iPS) cells are generated by retroviral or lentiviral transduction of reprogramming factors. Multiple viral integrations into the genome may cause insertional mutagenesis and may increase the risk of tumour formation. Non-integration methods have been reported to overcome the safety concerns associated with retro and lentiviral-derived iPS cells, such as transient expression of the reprogramming factors using episomal plasmids, and direct delivery of reprogramming mRNAs or proteins. In this review, we focus on the mechanisms of cellular reprogramming and current methods used to induce pluripotency. We also highlight problems associated with the generation of iPS cells. An increased understanding of the fundamental mechanisms underlying pluripotency and refining the methodology of iPS cell generation will have a profound impact on future development and application in regenerative medicine and reproductive biotechnology of farm animals.

• Reprogramming
• Large animal models
• Stemness
• Chimera
• Germline transmission
• Induced pluripotent stem cells
• Gene delivery

Cardiovascular disease, nervous system disorders, and cancer in association with other diseases such as diabetes mellitus result in greater than sixty percent of the global annual deaths. These noncommunicable diseases also affect at least one-third of the population in low and middle-income countries and lead to hypertension, elevated cholesterol, malignancy, and neurodegenerative disorders such as Alzheimer’s disease and stroke. With the climbing lifespan of the world’s population, increased prevalence of these disorders is expected requiring the development of new therapeutic strategies against these disabling disease entities. Targeting stem cell proliferation for cardiac disease, vascular disorders, cancer, and neurodegenerative disorders is receiving great enthusiasm, especially those that focus upon SIRT1, a mammalian homologue of the yeast silent information regulator-2. Modulation of the cellular activity of SIRT1 can involve oversight by nicotinamide/nicotinic acid mononucleotide adenylyltransferase, mammalian forkhead transcription factors, mechanistic of rapamycin pathways, and cysteine-rich protein 61, connective tissue growth factor, and nephroblastoma over-expressed gene family members that can impact cytoprotective outcomes. Ultimately, the ability of SIRT1 to control the programmed cell death pathways of apoptosis and autophagy can determine not only cardiac, vascular, and neuronal stem cell development and longevity, but also the onset of tumorigenesis and the resistance against chemotherapy. SIRT1 therefore has a critical role and holds exciting prospects for new therapeutic strategies that can offer reparative processes for cardiac, vascular, and nervous system degenerative disorders as well as targeted control of aberrant cell growth during cancer.

• FoxO
• Mechanistic of rapamycin
• Apoptosis
• Autophagy
• Cardiovascular
• Cysteine-rich protein 61, connective tissue growth factor, and nephroblastoma over-expressed gene
• Neurodegeneration
• Progenitor stem cells
• SIRT1
• Cancer

Cell transplantation is an attractive potential therapy for heart diseases. For example, myocardial infarction (MI) is a leading cause of mortality in many countries. Numerous medical interventions have been developed to stabilize patients with MI and, although this has increased survival rates, there is currently no clinically approved method to reverse the loss of cardiac muscle cells (cardiomyocytes) that accompanies this disease. Cell transplantation has been proposed as a method to replace cardiomyocytes, but a safe and reliable source of cardiogenic cells is required. An ideal source would be the patients’ own somatic tissue cells, which could be converted into cardiogenic cells and transplanted into the site of MI. However, these are difficult to produce in large quantities and standardized protocols to produce cardiac cells would be advantageous for the research community. To achieve these research goals, small molecules represent attractive tools to control cell behavior. In this editorial, we introduce the use of small molecules in stem cell research and summarize their application to the induction of cardiogenesis in non-cardiac cells. Exciting new developments in this field are discussed, which we hope will encourage cardiac stem cell biologists to further consider employing small molecules in their culture protocols.

• Cardiogenesis
• Cell reprogramming
• Somatic cells
• Small molecules
• Cardiovascular disease

Introduction. Dental pulp cells (DPCs) are promising cell source for dental tissue regeneration. Recently, small molecules which optimize microenvironment or activate the reprogramming network provide a new way to enhance the pluripotency. Two promising bioflavonoids luteolin and apigenin were reported to enhance reprogramming efficiency in mouse embryonic fibroblast (MEF). However, their effect and underlying mechanism in cell fate determination of human DPCs remain unclear. Methods. To elucidate the effect of luteolin and apigenin on the cell fate determination of DPCs, we explored the cell proliferation, cell cycle, senescence, apoptosis, expression of pluripotency markers Oct-4, Sox2, and c-Myc, and multilineage differentiation capability of DPCs with luteolin or apigenin treatment. Results. We demonstrated that luteolin and apigenin inhibited cell proliferation, arrested DPCs in G2/M and S phase, and upregulated PI value and apoptosis. Moreover, luteolin and apigenin increased telomerase activity, maintained DPCs in a presenescent state, and activated the expression of Oct-4, Sox2, and c-Myc at a dose- and time-dependent pattern in DPCs even at late passages, albeit repressed lineage-specific differentiation. Conclusions. Addition of luteolin and apigenin in the culture medium might provide an effective way to maintain DPCs in an undifferentiated stage and inhibit lineage-specific differentiation.

• RNA-sequencing
• age-related macular degeneration (AMD)
• cell-based transplantation therapy
• disease modelling
• drug screening
• induced pluripotent stem cells (iPSCs)
• retinal pigment epithelium (RPE)
• stem cells

• ampk
• apoptosis
• autophagy
• cardiac
• ccn
• diabetes mellitus
• erythropoietin
• forkhead
• foxo
• metformin
• nervous system
• oxidative stress
• mtor
• sirtuins
• sirt1
• stem cells
• vascular
• wisp1
• wnt

• Animals
• Apoptosis
• Autophagy
• Diabetes Mellitus
• Humans

• R01 NS053946 NINDS NIH HHS

• differentiation
• epigenetics
• metabolism
• mitochondria
• pluripotency

• Aurora kinase
• Cell transdifferentiation
• Chemical biology
• Indirubin
• Small molecules

Currently, there are genetic- and chemical-based methods for producing pluripotent stem cells from somatic cells, but all of them are extremely inefficient.  However, a simple and efficient technique has recently been reported by Obokata et al (2014a, b) that creates pluripotent stem cells through acid-based treatment of somatic cells.  These cells were named stimulus-triggered acquisition of pluripotency (STAP) stem cells. This would be a major game changer in regenerative medicine if the results could be independently replicated. Hence, we isolated CD45 ⁺ splenocytes from five-day-old Oct4-GFP mice and treated the cells with acidified (pH 5.7) Hank’s Balanced Salt Solution (HBSS) for 25 min, using the methods described by Obokata et al 2014c. However, we found that this method did not induce the splenocytes to express the stem cell marker Oct4-GFP when observed under a confocal microscope three to six days after acid treatment. qPCR analysis also confirmed that acid treatment did not induce the splenocytes to express the stemness markers Oct4, Sox2 and Nanog.  In addition, we obtained similar results from acid-treated Oct4-GFP lung fibroblasts. In summary, we have not been able to produce STAP stem cells from neonatal splenocytes or lung fibroblasts using the acid-based treatment reported by Obokata et al (2014a, b, c).

• child
• critical illness
• infant
• pediatric intensive care unit
• trauma