Manoj K. Gupta

Joslin Diabetes Center · Section on Islet Cell and Regenerative Biology

Non-alcoholic fatty liver disease (NAFLD) is a liver manifestation of metabolic syndrome, and is estimated to affect one billion individuals worldwide. An increased intake of a high-fat diet (HFD) and sugar-sweetened beverages are risk-factors for NAFLD development, but how their combined intake promotes progression to a more severe form of liver i...

MODY8 (maturity-onset diabetes of the young, type 8) is a dominantly inherited monogenic form of diabetes associated with mutations in the carboxyl ester lipase (CEL) gene expressed by pancreatic acinar cells. MODY8 patients develop childhood-onset exocrine pancreas dysfunction followed by diabetes during adulthood. However, it is unclear how CEL m...

Dysregulated glucagon secretion deteriorates glycemic control in type 1 and type 2 diabetes. Although insulin is known to regulate glucagon secretion via its cognate receptor (insulin receptor, INSR) in pancreatic alpha cells, the role of downstream proteins and signaling pathways underlying insulin’s activities are not fully defined. Using in vivo...

Human embryonic stem cells are a type of pluripotent stem cells (hPSCs) that are used to investigate their differentiation into diverse mature cell types for molecular studies. The mechanisms underlying insulin receptor (IR)-mediated signaling in the maintenance of human pluripotent stem cell (hPSC) identity and cell fate specification are not full...

The insulin and insulin-like growth factor-1 (IGF-1) receptors are important for the growth and development of embryonic tissues. To directly define their roles in the maintenance of pluripotency and differentiation of stem cells, we knocked out both the receptors in induced pluripotent stem cells (iPSCs). iPSCs lacking both the insulin and IGF-1 r...

The role of leptin receptor (OB-R) signaling in linking pluripotency with growth and development and the consequences of dysfunctional leptin signaling on progression of metabolic disease is poorly understood. Using a global unbiased proteomics approach we report that embryonic fibroblasts (MEFs) carrying the db/db mutation exhibit metabolic abnorm...

Dietary sugars, fructose and glucose, promote hepatic de novo lipogenesis and modify the effects of a high-fat diet (HFD) on the development of insulin resistance. Here, we show that fructose and glucose supplementation of an HFD exert divergent effects on hepatic mitochondrial function and fatty acid oxidation. This is mediated via three different...

The regulation of islet cell biology is critical for glucose homeostasis1. N6-methyladenosine (m6A) is the most abundant internal messenger RNA (mRNA) modification in mammals2. Here, we report that the m6A landscape segregates human type 2 diabetes (T2D) islets from controls significantly better than the transcriptome and that m6A is vital for β-ce...

A major target of insulin signaling is the FoxO family of Forkhead transcription factors, which translocate from the nucleus to the cytoplasm following insulin-stimulated phosphorylation. Here we show that the Forkhead transcription factors FoxK1 and FoxK2 are also downstream targets of insulin action, but that following insulin stimulation, they t...

Solute Carrier Family 19 Member 2 (SLC19A2) encodes thiamine transporter 1 (THTR1), which facilitates thiamine transport across the cell membrane. SLC19A2 homozygous mutations have been described as a cause of thiamine-responsive megaloblastic anemia (TRMA), an autosomal recessive syndrome characterized by megaloblastic anemia, diabetes, and sensor...

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A finely tuned balance of self-renewal, differentiation, proliferation, and survival governs the pool size and regenerative capacity of blood-forming hematopoietic stem and progenitor cells (HSPCs). Here, we report that protein kinase C delta (PKCδ) is a critical regulator of adult HSPC number and function that couples the proliferative and metabol...

Objectives Insulin receptor (IR)-mediated signaling is involved in the regulation of pluripotent stem cells; however, its direct effects on regulating the maintenance of pluripotency and lineage development are not fully understood. The main objective of this study is to understand the role of IR signaling in pluripotency and lineage development....

Pluripotent stem cells (PSCs) express insulin receptors (IR) and insulin-like growth factor-1 receptors (IGF-1R). In human embryonic stem cells (hESCs), IGF-1R is necessary for cell survival, self-renewal and differentiation into endoderm. Data on the role of IR in hESCs is limited; it’s been linked to self-renewal and pluripotency of murine neural...

Objectives Insulin receptor (IR)-mediated signaling is involved in the regulation of pluripotent stem cells; however, its direct effects on regulating the maintenance of pluripotency and lineage development are not fully understood. The main objective of this study is to understand the role of IR signaling in pluripotency and lineage development....

Overconsumption of high-fat diet (HFD) and sugar-sweetened beverages are risk factors for developing obesity, insulin resistance, and fatty liver disease. Here we have dissected mechanisms underlying this association using mice fed either chow or HFD with or without fructose- or glucose-supplemented water. In chow-fed mice, there was no major physi...

A major goal of diabetes research is to develop strategies that replenish pancreatic insulin-producing beta cells. One emerging strategy is to harness pancreatic plasticity—the ability of pancreatic cells to undergo cellular interconversions—a phenomenon implicated in physiological stress and pancreatic injury. Here, we investigate the effects of i...

The potential use of stem cells for cell-based tissue repair and regeneration offers alternative therapeutic strategies for various diseases. Adipose-derived stem cells (ADSCs) have emerged as a promising source of stem cells suitable for transplantation in regenerative medicine and wound repair. A recent publication in Stem Cell Research & Therapy...

: The impact of somatic cell proliferation rate on induction of pluripotent stem cells remains controversial. Herein, we report that rapid proliferation of human somatic fibroblasts is detrimental to reprogramming efficiency when reprogrammed using a lentiviral vector expressing OCT4, SOX2, KLF4, and cMYC in insulin-rich defined medium. Human fibro...

The mechanisms underlying the development of complications in type 1 diabetes (T1D) are poorly understood. Disease modeling of induced pluripotent stem cells (iPSCs) from patients with longstanding T1D (disease duration ≥ 50 years) with severe (Medalist +C) or absent to mild complications (Medalist -C) revealed impaired growth, reprogramming, and d...

Background: Diabetes and metabolic syndromes are chronic, devastating diseases with increasing prevalence. Human pluripotent stem cells are gaining popularity in their usage for human in vitro disease modeling. With recent rapid advances in genome editing tools, these cells can now be genetically manipulated with relative ease to study how genes a...

Blood formation by hematopoietic stem cells (HSCs) is regulated by a still incompletely defined network of general and HSC-specific regulators. In this study, we analyzed the role of G-protein coupled receptor 56 (Gpr56) as a candidate HSC regulator based on its differential expression in quiescent relative to proliferating HSCs and its common targ...

MicroRNAs regulate target gene expression post-transcriptionally in a myriad of cell types and play critical roles in diverse physiological and pathological processes, including cardiomyocyte development, differentiation, and regeneration. The recent publication in Stem Cell Research and Therapy by Wagh and colleagues reports a novel regulatory rol...

MicroRNAs regulate target gene expression post-transcriptionally in a myriad of cell types and play critical roles in diverse physiological and pathological processes, including cardiomyocyte development, differentiation, and regeneration. The recent publication in Stem Cell Research and Therapy by Wagh and colleagues reports a novel regulatory rol...

Long QT syndromes (LQTS) are heritable diseases characterized by prolongation of the QT interval on an electrocardiogram, which often leads to syncope and sudden cardiac death. Here we report the generation of induced pluripotent stems (iPS) cells from two patients with LQTS type 3 carrying a different point mutation in a sodium channel Nav1.5 (p.V...

Circulating immune complexes (CICs) generated during a variety of diseased conditions, comprise aggregates of immunoglobulin molecules and trapped antigens. Aggregated antibodies in CICs present multiple cross-linked Fc regions of antibodies which can potentially bind low-affinity Fc receptors (FcRs) expressed on a variety of leukocyte populations....

Dihydropyridines (DHP), which nifedipine is a member of, preferentially block Ca(2+) channels of different cell types. Moreover, influx of Ca(2+) through L-type Ca(2+) channels (LTCCs) activates Ca(2+) signaling pathways, which in turn contribute to numerous cellular processes. Although LTCCs are expressed in undifferentiated cells, very little is...

List of samples used for global gene expression profiling. (TIF)

Primer sequences used for normal and quantitative RT-PCR analysis of total RNA isolated from ES cell-derived CMs cultured under control and nifedipine-treated conditions. Abbreviations: ANF/Nppa, natriuretic peptide precursor A; ES cell, embryonic stem cell; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; α-MHC/Myh6, myosin heavy chain 6, MLC2v/My...

(A) Hierarchical clustering and (B) Principal component analysis of variable genes expression during induction of CMs under control and nifedipine-treated conditions at day 4 and 12 of differentiation. (C–D) Proliferation and viability assay of untreated and nifedipine-treated ES cells. Nifedipine did not alter cell proliferation (C) or viability (...

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Representative EB showing EGFP-positive CMs differentiated under control conditions. (MPG)

After nifedipine treatment. The videos show the 3D structure of the entire EB. (MPG)

(A) Representative eGFP+ CMs obtained from dissociated EBs cultured under control conditions or in the presence of nifedipine or BayK8644. (B) eGFP-positive cells (CMs) at day 12 of differentiation generated under nifedipine- and BayK8644-treatment. CMs were manually counted and are expressed as a percentage of total CMs generated under control con...

Representative Ca2+ transients obtained from control and nifedipine-treated CMs. (A) Developmental changes of spontaneous Ca2+ transients of cells generated from EBs at day 2, 4, 6, 8, and 10 of differentiation. Note the time-dependent increase of the Ca2+ transients’ amplitude of cell generated under control and nifedipine-treated conditions. (B)...

Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is an epigenetic phenomenon. It has been suggested that iPSC retain some tissue-specific memory whereas little is known about interindividual epigenetic variation. We have reprogrammed mesenchymal stromal cells from human bone marrow (iP-MSC) and compared their DNA methylati...

Functional and molecular integrity of cardiomyocytes (CMs) derived from induced pluripotent stem (iPS) cells is essential for their use in tissue repair, disease modelling and drug screening. In this study we compared global transcriptomes of beating clusters (BCs) microdissected from differentiating human iPS cells and embryonic stem (ES) cells. H...

Collection of Figures S1-S3, Tables S1-S5, Supplemental results, methods and references. This file is a PDF file containing Figure S1 (characterization of pluripotency markers in undifferentiated human iPS and ES cells), Figure S2 (assessment of the cardiogenic potential of human iPS and ES cells), Figure S3 (expression of pluripotency genes in bea...

List of genes differentially expressed between undifferentiated iPS and ES cells. This Excel file contains Tables S7 and S8 with the list of genes that are, respectively, upregulated and downregulated in undifferentiated iPS cells compared to ES cells.

qRT-PCR validation of microarray data with Human ES cell StellArray qPCR Array (Lonza). This file is a PDF document containing Table S15 showing the results of qRT-PCR analysis of expression of selected pluripotency and differentiations genes in undifferentiated human ES and iPS cells in comparison with microarray data obtained in this study.

List of differentially expressed genes in iPS-BC versus iPS cell comparison and ES-BC versus ES cell comparison. This file is an Excel file containing Table S16 (list of 1235 genes upregulated in iPS-BC compared to iPS cells), Table S17 (list of 1014 genes downregulated in iPS-BC compared to iPS cells), Table S18 (list of 875 genes upregulated in E...

List of genes that are differentially expressed across all samples. This file is an Excel file containing Table S6 with the list of 4254 variable genes and their expression levels in all samples. Variable genes were defined by a coefficient of variation (SD/mean) between 0.5-10.

List of genes differentially expressed between iPS cell- and ES cell-derived beating clusters (BCs). This Excel file contains Tables S9 and S10 with the list of genes that are, respectively, upregulated and downregulated in iPS-BCs compared to ES-BCs.

Expression levels of selected human ES cell- and cardiacspecific or enriched genes across all samples. This is a PDF file in a landscape format containing Table S12 (list of 27 ES cell-specific genes and their expression levels across all samples), Table S13 (list of 49 human ES cell-enriched genes with their expression levels in all samples) and T...

Expression levels of selected genes included in GO-terms that were found to be overrepresented in iPS-BCs compared to ES-BCs. This PDF file contains the Table S11 displaying the expression levels of selected fibroblastassociated genes found to be overexpressed in iPS-BCs compared to ES-BCs.

Gene ontology analysis of genes upregulated in iPS-BCs and ES-BCs as compared to their respective undifferentiated counterparts. This file is a PDF document containing Table S20 with the list of all overrepresented GO terms in iPS-BCs compared to iPS cells and in ES-BCs compared to ES cells.

Stable insertion of selection markers. Integration of SV40-Neo was probes with primers specific for the SV40 promoter and the Neomycin resistance (a). Primers for the UTF1-promoter driven Neomycin resistance were used to test for stable integration of the UTF1-Neo transgene (b). (1.29 MB TIF)

Expression of viral Oct4 in UTF-Neo-selected and non-selected cells. a) Relative expressions of the viral oct-4 gene were assayed by real time PCR. Normalized for the expression of viral Oct4 in the selected clone UTF-1 passage 4 the relative expression level of viral Oct4 are shown for later passages (7, 11) of UTF-1 and for non-selected TiB7-4 iP...

Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells. Recently, selection of fully reprogrammed cells was achieved based on colony morphology reminiscent of embryonic stem (ES) cells. The maintenance of pluripotency was analysed. Clonal murine iPS cell line TiB7-4, which was derived from adult fibroblasts, was analysed for ma...

Characterisation of potential antigen(s) in the teliospore wall of Tilletia indica, the causal agent of Karnal bunt (KB) of wheat (Triticum aestivum) is necessary to generate specific immunoprobes for the development of reliable immunodiagnostic procedures. The intact teliospores of KB were used to generate anti-teliospore antibodies for the develo...

Embryonic, fetal, and adult stem cells have been extensively tested in the past decade as candidates for cell replacement therapy of severe heart failure. Among them, embryonic stem (ES) cells have been regarded as an especially valuable source of therapeutic cells due to their unlimited growth in culture and well-established cardiogenic potential...