The authors of the article titled "Advances in Genetic Reprogramming: Prospects from Developmental Biology to Regenerative Medicine" (Dhanjal DS, Singh R, Sharma V, Nepovimova E, Adam V, Kuca K, Chopra C. Curr Med Chem. 2024; 31(13): 1646-1690. DOI: 10.2174/0929867330666230503144619. PMID: 37138422) [1] have made revisions to the references in the text and the reference section. These updates have been made to ensure the integrity of the article. The updated reference list can be found in the latest version of the article. The authors apologize for any confusion or inconvenience caused. The original article can be found online at: https://www.eurekaselect.com/article/131443.
- MeSH
- Humans MeSH
- Cellular Reprogramming * genetics MeSH
- Regenerative Medicine * MeSH
- Developmental Biology MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Aim: Human induced pluripotent stem cells (iPSCs) are inefficiently derived from somatic cells by overexpression of defined transcription factors. Overexpression of H2A histone variant macroH2A1.1, but not macroH2A1.2, leads to increased iPSC reprogramming by unclear mechanisms. Materials & methods: Cleavage under targets and tagmentation (CUT&Tag) allows robust epigenomic profiling of a low cell number. We performed an integrative CUT&Tag-RNA-Seq analysis of macroH2A1-dependent orchestration of iPSCs reprogramming using human endothelial cells. Results: We demonstrate wider genome occupancy, predicted transcription factors binding, and gene expression regulated by macroH2A1.1 during reprogramming, compared to macroH2A1.2. MacroH2A1.1, previously associated with neurodegenerative pathologies, specifically activated ectoderm/neural processes. Conclusion: CUT&Tag and RNA-Seq data integration is a powerful tool to investigate the epigenetic mechanisms occurring during cell reprogramming.
- MeSH
- Endothelial Cells metabolism MeSH
- Histones * metabolism MeSH
- Induced Pluripotent Stem Cells * metabolism MeSH
- Humans MeSH
- Cellular Reprogramming genetics MeSH
- RNA-Seq MeSH
- Transcription Factors genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
One of the challenges in clinical translation of cell-replacement therapies is the definition of optimal cell generation and storage/recovery protocols which would permit a rapid preparation of cell-treatment products for patient administration. Besides, the availability of injection devices that are simple to use is critical for potential future dissemination of any spinally targeted cell-replacement therapy into general medical practice. Here, we compared the engraftment properties of established human-induced pluripotent stem cells (hiPSCs)-derived neural precursor cell (NPCs) line once cells were harvested fresh from the cell culture or previously frozen and then grafted into striata or spinal cord of the immunodeficient rat. A newly developed human spinal injection device equipped with a spinal cord pulsation-cancelation magnetic needle was also tested for its safety in an adult immunosuppressed pig. Previously frozen NPCs showed similar post-grafting survival and differentiation profile as was seen for freshly harvested cells. Testing of human injection device showed acceptable safety with no detectable surgical procedure or spinal NPCs injection-related side effects.
- MeSH
- Cell Differentiation physiology MeSH
- Adult MeSH
- Genetic Vectors genetics MeSH
- Induced Pluripotent Stem Cells * physiology transplantation MeSH
- Rats MeSH
- Humans MeSH
- Spinal Cord MeSH
- Brain MeSH
- Neural Stem Cells * physiology transplantation MeSH
- Specimen Handling methods MeSH
- Tissue and Organ Harvesting methods MeSH
- Swine MeSH
- Cellular Reprogramming * genetics physiology MeSH
- Graft Survival physiology MeSH
- Injections, Spinal * adverse effects instrumentation methods MeSH
- Stem Cell Transplantation * adverse effects instrumentation methods MeSH
- Sendai virus MeSH
- Treatment Outcome MeSH
- Animals MeSH
- Check Tag
- Adult MeSH
- Rats MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Embryonic stem cells and induced pluripotent stem cells provided us with fascinating new knowledge in recent years. Mechanistic insight into intricate regulatory circuitry governing pluripotency stemness and disclosing parallels between pluripotency stemness and cancer instigated numerous studies focusing on roles of pluripotency transcription factors, including Oct4, Sox2, Klf4, Nanog, Sall4 and Tfcp2L1, in cancer. Although generally well substantiated as tumour-promoting factors, oncogenic roles of pluripotency transcription factors and their clinical impacts are revealing themselves as increasingly complex. In certain tumours, both Oct4 and Sox2 behave as genuine oncogenes, and reporter genes driven by composite regulatory elements jointly recognized by both the factors can identify stem-like cells in a proportion of tumours. On the other hand, cancer stem cells seem to be biologically very heterogeneous both among different tumour types and among and even within individual tumours. Pluripotency transcription factors are certainly implicated in cancer stemness, but do not seem to encompass its entire spectrum. Certain cancer stem cells maintain their stemness by biological mechanisms completely different from pluripotency stemness, sometimes even by engaging signalling pathways that promote differentiation of pluripotent stem cells. Moreover, while these signalling pathways may well be antithetical to stemness in pluripotent stem cells, they may cooperate with pluripotency factors in cancer stem cells - a paradigmatic example is provided by the MAPK-AP-1 pathway. Unexpectedly, forced expression of pluripotency transcription factors in cancer cells frequently results in loss of their tumour-initiating ability, their phenotypic reversion and partial epigenetic normalization. Besides the very different signalling contexts operating in pluripotent and cancer stem cells, respectively, the pronounced dose dependency of reprogramming pluripotency factors may also contribute to the frequent loss of tumorigenicity observed in induced pluripotent cancer cells. Finally, contradictory cell-autonomous and non-cell-autonomous effects of various signalling molecules operate during pluripotency (cancer) reprogramming. The effects of pluripotency transcription factors in cancer are thus best explained within the concept of cancer stem cell heterogeneity.
- MeSH
- Cell Differentiation genetics MeSH
- Embryonic Stem Cells MeSH
- Induced Pluripotent Stem Cells * metabolism MeSH
- Humans MeSH
- Neoplasms * genetics metabolism MeSH
- Octamer Transcription Factor-3 genetics metabolism MeSH
- Pluripotent Stem Cells * MeSH
- Cellular Reprogramming genetics MeSH
- Transcription Factors genetics metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Significance: Since their discovery, induced pluripotent stem cells (iPSCs) had generated considerable interest in the scientific community for their great potential in regenerative medicine, disease modeling, and cell-based therapeutic approach, due to their unique characteristics of self-renewal and pluripotency. Recent Advances: Technological advances in iPSC genome-wide epigenetic profiling led to the elucidation of the epigenetic control of cellular identity during nuclear reprogramming. Moreover, iPSC physiology and metabolism are tightly regulated by oxidation-reduction events that mainly occur during the respiratory chain. In theory, iPSC-derived differentiated cells would be ideal for stem cell transplantation as autologous cells from donors, as the risks of rejection are minimal. Critical Issues: However, iPSCs experience high oxidative stress that, in turn, confers a high risk of increased genomic instability, which is most often linked to DNA repair deficiencies. Genomic instability has to be assessed before iPSCs can be used in therapeutic designs. Future Directions: This review will particularly focus on the links between redox balance and epigenetic modifications-in particular based on the histone variant macroH2A1-that determine DNA damage response in iPSCs and derived differentiated cells, and that might be exploited to decrease the teratogenic potential on iPSC transplantation. Antioxid. Redox Signal. 34, 335-349.
- MeSH
- Cell Differentiation * genetics MeSH
- Cell Self Renewal MeSH
- Epigenesis, Genetic * MeSH
- Induced Pluripotent Stem Cells cytology metabolism MeSH
- Humans MeSH
- DNA Methylation MeSH
- Mitochondria genetics metabolism MeSH
- Cell Transformation, Neoplastic genetics metabolism MeSH
- Genomic Instability MeSH
- Oxidation-Reduction * MeSH
- Oxidative Stress MeSH
- Oxidative Phosphorylation MeSH
- Pluripotent Stem Cells cytology metabolism MeSH
- Cellular Reprogramming genetics MeSH
- Regenerative Medicine MeSH
- Stem Cell Transplantation MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Histone H3.3 glycine 34 to arginine/valine (G34R/V) mutations drive deadly gliomas and show exquisite regional and temporal specificity, suggesting a developmental context permissive to their effects. Here we show that 50% of G34R/V tumors (n = 95) bear activating PDGFRA mutations that display strong selection pressure at recurrence. Although considered gliomas, G34R/V tumors actually arise in GSX2/DLX-expressing interneuron progenitors, where G34R/V mutations impair neuronal differentiation. The lineage of origin may facilitate PDGFRA co-option through a chromatin loop connecting PDGFRA to GSX2 regulatory elements, promoting PDGFRA overexpression and mutation. At the single-cell level, G34R/V tumors harbor dual neuronal/astroglial identity and lack oligodendroglial programs, actively repressed by GSX2/DLX-mediated cell fate specification. G34R/V may become dispensable for tumor maintenance, whereas mutant-PDGFRA is potently oncogenic. Collectively, our results open novel research avenues in deadly tumors. G34R/V gliomas are neuronal malignancies where interneuron progenitors are stalled in differentiation by G34R/V mutations and malignant gliogenesis is promoted by co-option of a potentially targetable pathway, PDGFRA signaling.
- MeSH
- Astrocytes metabolism pathology MeSH
- Models, Biological MeSH
- Cell Lineage MeSH
- Chromatin metabolism MeSH
- Embryo, Mammalian metabolism MeSH
- Epigenesis, Genetic MeSH
- Transcription, Genetic MeSH
- Glioma genetics pathology MeSH
- Histones genetics metabolism MeSH
- Interneurons metabolism MeSH
- Carcinogenesis genetics pathology MeSH
- Lysine metabolism MeSH
- Mutation genetics MeSH
- Mice, Inbred C57BL MeSH
- Brain Neoplasms genetics pathology MeSH
- Neural Stem Cells metabolism MeSH
- Oligodendroglia metabolism MeSH
- Prosencephalon embryology MeSH
- Cellular Reprogramming genetics MeSH
- Promoter Regions, Genetic genetics MeSH
- Gene Expression Regulation, Neoplastic MeSH
- Receptor, Platelet-Derived Growth Factor alpha genetics metabolism MeSH
- Neoplasm Grading MeSH
- Transcriptome genetics MeSH
- Gene Silencing MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Reprogramming of non-endocrine pancreatic cells into insulin-producing cells represents a promising therapeutic approach for the restoration of endogenous insulin production in diabetic patients. In this paper, we report that human organoid cells derived from the pancreatic tissue can be reprogrammed into the insulin-producing cells (IPCs) by the combination of in vitro transcribed modified mRNA encoding transcription factor neurogenin 3 and small molecules modulating the epigenetic state and signalling pathways. Upon the reprogramming, IPCs formed 4.6 ± 1.2 % of the total cells and expressed typical markers (insulin, glucokinase, ABCC8, KCNJ11, SLC2A2, SLC30A8) and transcription factors (PDX1, NEUROD1, MAFA, NKX2.2, NKX6.1, PAX4, PAX6) needed for the proper function of pancreatic β-cells. Additionally, we have revealed a positive effect of ALK5 inhibitor RepSox on the overall reprogramming efficiency. However, the reprogrammed IPCs possessed only a partial insulin-secretory capacity, as they were not able to respond to the changes in the extracellular glucose concentration by increasing insulin secretion. Based on the achieved results we conclude that due to the incomplete reprogramming, the IPCs have immature character and only partial properties of native human β-cells.
- MeSH
- AC133 Antigen metabolism MeSH
- Insulin-Secreting Cells cytology drug effects MeSH
- Adult MeSH
- Transcription, Genetic drug effects MeSH
- Insulin biosynthesis MeSH
- Small Molecule Libraries pharmacology MeSH
- Humans MeSH
- RNA, Messenger genetics metabolism MeSH
- Organoids cytology MeSH
- Cellular Reprogramming drug effects genetics MeSH
- Cell Proliferation MeSH
- Nerve Tissue Proteins genetics metabolism MeSH
- Basic Helix-Loop-Helix Transcription Factors genetics metabolism MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- MeSH
- Cell Differentiation physiology genetics MeSH
- Chimera genetics metabolism MeSH
- Embryonic Stem Cells cytology physiology MeSH
- Induced Pluripotent Stem Cells * cytology physiology ultrastructure MeSH
- Models, Animal MeSH
- Mice, Transgenic MeSH
- Cellular Reprogramming physiology genetics MeSH
- Cellular Reprogramming Techniques methods MeSH
- In Vitro Techniques MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
MicroRNA (miRNAs) are short noncoding RNA molecules involved in many cellular processes and shown to play a key role in somatic cell induced reprogramming. We performed an array based screening to identify candidates that are differentially expressed between dermal skin fibroblasts (DFs) and induced pluripotent stem cells (iPSCs). We focused our investigations on miR-145 and showed that this candidate is highly expressed in DFs relative to iPSCs and significantly downregulated during reprogramming process. Inhibition of miR-145 in DFs led to the induction of "cellular plasticity" demonstrated by: (a) alteration of cell morphology associated with downregulation of mesenchymal and upregulation of epithelial markers; (b) upregulation of pluripotency-associated genes including SOX2, KLF4, C-MYC; (c) downregulation of miRNA let-7b known to inhibit reprogramming; and (iv) increased efficiency of reprogramming to iPSCs in the presence of reprogramming factors. Together, our results indicate a direct functional link between miR-145 and molecular pathways underlying reprogramming of somatic cells to iPSCs.
- MeSH
- Fibroblasts cytology metabolism MeSH
- Induced Pluripotent Stem Cells cytology MeSH
- Humans MeSH
- MicroRNAs genetics metabolism MeSH
- Molecular Sequence Data MeSH
- Cellular Reprogramming * genetics MeSH
- Gene Expression Regulation MeSH
- Reproducibility of Results MeSH
- Base Sequence MeSH
- Dermis cytology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The potential clinical applications of human induced pluripotent stem cells (hiPSCs) are limited by genetic and epigenetic variations among hiPSC lines and the question of their equivalency with human embryonic stem cells (hESCs). We used MethylScreen technology to determine the DNA methylation profile of pluripotency and differentiation markers in hiPSC lines from different source cell types compared to hESCs and hiPSC source cells. After derivation, hiPSC lines compromised a heterogeneous population characterized by variable levels of aberrant DNA methylation. These aberrations were induced during somatic cell reprogramming and their levels were associated with the type of hiPSC source cells. hiPSC population heterogeneity was reduced during prolonged culture and hiPSCs acquired an hESC-like methylation profile. In contrast, the expression of differentiation marker genes in hiPSC lines remained distinguishable from that in hESCs. Taken together, in vitro culture facilitates hiPSC acquisition of hESC epigenetic characteristics. However, differences remain between both pluripotent stem cell types, which must be considered before their use in downstream applications.
- MeSH
- Cell Differentiation genetics MeSH
- Cell Line MeSH
- Fibroblasts cytology metabolism MeSH
- Induced Pluripotent Stem Cells cytology metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Human Embryonic Stem Cells cytology metabolism MeSH
- DNA Methylation * MeSH
- Cellular Reprogramming genetics MeSH
- Cluster Analysis MeSH
- Gene Expression Profiling MeSH
- Gene Expression Regulation, Developmental MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
