scholarly journals Direct conversion of human fibroblasts into osteoblasts and osteocytes with small molecules and a single factor, Runx2

2017 ◽  
Author(s):  
Yanjiao Li ◽  
YaoLong Wang ◽  
Juehua Yu ◽  
Zhaoxia Ma ◽  
Qiong Bai ◽  
...  
Keyword(s):  
Small Molecules ◽  
Human Fibroblasts ◽  
Direct Conversion ◽  
Direct Reprogramming ◽  
Cell Type ◽  
Dynamic Nature ◽  
Post Transplantation ◽  
Genetic Manipulations ◽  
Cell Based Therapy ◽  
Promising Strategy

AbstractHuman osteoblasts can be induced from somatic cells by introducing defined factors, however, the strategy limits cells therapeutic applications for its multi-factor and complicated genetic manipulations that may bring uncertainty into the genome. Another important cell type in bone metabolism, osteocytes, which play a central role in regulating the dynamic nature of bone in all its diverse functions, have not been obtained from transdifferetiation so far. Herein, we have established procedures to convert human fibroblast directly into osteocyte-like and osteoblast-like cells using a single transcription factor, Runx2 and chemical cocktails by activating Wnt and cAMP/PKA pathways. These induced osteoblast-like cells express osteogenic markers and generate mineralized nodule deposition. A good performance of bone formation from these cells was observed in subcutaneous site of mouse at 4 weeks post-transplantation. Moreover, further studies convert human fibroblasts into osteocyte-like cells by orchestrating timing of the aforementioned chemical cocktails exposure. These osteocyte-like cells express osteocyte-specific markers and display characteristic morphology features of osteocytes. In summary, this study provides a promising strategy for cell-based therapy in bone regenerative medicine by direct reprogramming of fibroblasts into osteocytes and osteoblasts.

scholarly journals Highly efficient direct conversion of human monocytes into neuronal cells using a small molecule combination

2019 ◽  
Author(s):  
Itaru Ninomiya ◽  
Masato Kanazawa ◽  
Akihide Koyama ◽  
Masahiro Hatakeyama ◽  
Osamu Onodera
Keyword(s):  
Small Molecules ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Human Fibroblasts ◽  
Neuronal Cells ◽  
Neuronal Cell ◽  
Direct Conversion ◽  
Dermal Fibroblasts ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes

SummaryPrevious studies reported that human fibroblasts and astrocytes were successfully converted into neuronal cells by small molecules without introducing ectopic transgenes. Induced neuronal cells—reprogrammed directly from dermal fibroblasts or brain astrocytes—were obtained from some donors; however, the clinical applications of this approach would be limited because it requires an invasive biopsy to harvest enough cells for derivation. Here, we report that adult human peripheral blood monocytes may be directly converted into neuron-like cells using only a combination of small molecules without transgene integration. This method enables neuronal cell generation from TUJ1-positive cells after 3 days of induction (at over 80% conversion efficacy). These cells presented neuronal morphologies and markers, suggesting that terminally differentiated human cells may be efficiently transdifferentiated into a distantly related lineage. Overall, our study provides a strategy to develop neuronal cells directly from human adult peripheral blood monocytes using a generate transgene-free, chemical-only approach.

scholarly journals Direct Conversion of Normal and Alzheimer’s Disease Human Fibroblasts into Neuronal Cells by Small Molecules

2015 ◽  
Vol 17 (2) ◽  
pp. 204-212 ◽  
Author(s):  
Wenxiang Hu ◽  
Binlong Qiu ◽  
Wuqiang Guan ◽  
Qinying Wang ◽  
Min Wang ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Small Molecules ◽  
Human Fibroblasts ◽  
Neuronal Cells ◽  
Direct Conversion

scholarly journals Placenta to cartilage: direct conversion of human placenta to chondrocytes with transformation by defined factors

2012 ◽  
Vol 23 (18) ◽  
pp. 3511-3521 ◽  
Author(s):  
Ryuga Ishii ◽  
Daisuke Kami ◽  
Masashi Toyoda ◽  
Hatsune Makino ◽  
Satoshi Gojo ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Human Fibroblasts ◽  
Direct Conversion ◽  
Cellular Reprogramming ◽  
Irreversible Processes ◽  
Cell Based Therapy ◽  
Decidual Cells ◽  
Cell Conversion

Cellular differentiation and lineage commitment are considered to be robust and irreversible processes during development. Recent work has shown that mouse and human fibroblasts can be reprogrammed to a pluripotent state with a combination of four transcription factors. We hypothesized that combinatorial expression of chondrocyte-specific transcription factors could directly convert human placental cells into chondrocytes. Starting from a pool of candidate genes, we identified a combination of only five genes (5F pool)—BCL6, T (also called BRACHYURY), c-MYC, MITF, and BAF60C (also called SMARCD3)—that rapidly and efficiently convert postnatal human chorion and decidual cells into chondrocytes. The cells generated expressed multiple cartilage-specific genes, such as Collagen type II α1, LINK PROTEIN-1, and AGGRECAN, and exhibited characteristics of cartilage both in vivo and in vitro. Expression of the endogenous genes for T and MITF was initiated, implying that the cell conversion is due to not only the forced expression of the transgenes, but also to cellular reprogramming by the transgenes. This direct conversion system from noncartilage tissue to cartilaginous tissue is a substantial advance toward understanding cartilage development, cell-based therapy, and oncogenesis of chondrocytes.

scholarly journals Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Hua Qin ◽  
An-Dong Zhao ◽  
Meng-Li Sun ◽  
Kui Ma ◽  
Xiao-Bing Fu
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Stem Cell ◽  
Small Molecules ◽  
Human Fibroblasts ◽  
Intermediate Stage ◽  
Neural Progenitor ◽  
Chemical Induction ◽  
Electrophysiological Properties ◽  
Cell Based Therapy

Abstract Background Generation of neurons is essential in cell replacement therapy for neurodegenerative disorders like Parkinson’s disease. Several studies have reported the generation of dopaminergic (DA) neurons from mouse and human fibroblasts by ectopic expression of transcription factors, in which genetic manipulation is associated with potential risks. Methods The small molecules and protein factors were selected based on their function to directly induce human fetal lung IMR-90 fibroblasts into DA neuron-like cells. Microscopical, immunocytochemical, and RT-qPCR analyses were used to characterize the morphology, phenotype, and gene expression features of the induced cells. The whole-cell patch-clamp recordings were exploited to measure the electrophysiological properties. Results Human IMR-90 fibroblasts were rapidly converted into DA neuron-like cells after the chemical induction using small molecules and protein factors, with a yield of approximately 95% positive TUJ1-positive cells. The induced DA neuron-like cells were immunopositive for pan-neuronal markers MAP2, NEUN, and Synapsin 1 and DA markers TH, DDC, DAT, and NURR1. The chemical induction process did not involve a neural progenitor/stem cell intermediate stage. The induced neurons could fire single action potentials, which reflected partially the electrophysiological properties of neurons. Conclusion We developed a chemical cocktail of small molecules and protein factors to convert human fibroblasts into DA neuron-like cells without passing through a neural progenitor/stem cell intermediate stage. The induced DA neuron-like cells from human fibroblasts might provide a cellular source for cell-based therapy of Parkinson’s disease in the future.

scholarly journals STEM-20. A CANCER STEM CELL-SELECTIVE APOPTOSIS-INDUCING SMALL MOLECULE FOR THE TREATMENT OF GBM

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii200-ii200
Author(s):  
Stephen Skirboll ◽  
Natasha Lucki ◽  
Genaro Villa ◽  
Naja Vergani ◽  
Michael Bollong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Small Molecules ◽  
Small Molecule ◽  
Large Scale ◽  
Critical Role ◽  
Tumor Formation ◽  
Cell Type ◽  
Caspase 1 ◽  
Activation Assay

Abstract INTRODUCTION Glioblastoma multiforme (GBM) is the most aggressive form of primary brain cancer. A subpopulation of multipotent cells termed GBM cancer stem cells (CSCs) play a critical role in tumor initiation and maintenance, drug resistance, and recurrence following surgery. New therapeutic strategies for the treatment of GBM have recently focused on targeting CSCs. Here we have used an unbiased large-scale screening approach to identify drug-like small molecules that induce apoptosis in GBM CSCs in a cell type-selective manner. METHODS A luciferase-based survival assay of patient-derived GBM CSC lines was established to perform a large-scale screen of ∼one million drug-like small molecules with the goal of identifying novel compounds that are selectively toxic to chemoresistant GBM CSCs. Compounds found to kill GBM CSC lines as compared to control cell types were further characterized. A caspase activation assay was used to evaluate the mechanism of induced cell death. A xenograft animal model using patient-derived GBM CSCs was employed to test the leading candidate for suppression of in vivo tumor formation. RESULTS We identified a small molecule, termed RIPGBM, from the cell-based chemical screen that induces apoptosis in primary patient-derived GBM CSC cultures. The cell type-dependent selectivity of RIPGBM appears to arise at least in part from redox-dependent formation of a proapoptotic derivative, termed cRIPGBM, in GBM CSCs. cRIPGBM induces caspase 1-dependent apoptosis by binding to receptor-interacting protein kinase 2 (RIPK2) and acting as a molecular switch, which reduces the formation of a prosurvival RIPK2/TAK1 complex and increases the formation of a proapoptotic RIPK2/caspase 1 complex. In an intracranial GBM xenograft mouse model, RIPGBM was found to significantly suppress tumor formation. CONCLUSIONS Our chemical genetics-based approach has identified a small molecule drug candidate and a potential drug target that selectively targets cancer stem cells and provides an approach for the treatment of GBMs.

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