scholarly journals Reprogramming of Mice Primary Hepatocytes into Insulin-Producing Cells by Transfection with Multicistronic Vectors

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Haizhao Luo ◽  
Rongping Chen ◽  
Rui Yang ◽  
Yan Liu ◽  
Youping Chen ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Transcription Factor ◽  
Ectopic Expression ◽  
Potential Method ◽  
Diabetic Mice ◽  
Primary Hepatocytes ◽  
Neurogenin 3 ◽  
Transfected Cells ◽  
Insulin Producing Cells

The neogenesis of insulin-producing cells (IPCs) from non-beta-cells has emerged as a potential method for treating diabetes mellitus (DM). Many groups have documented that activation of pancreatic transcription factor(s) in hepatocytes can improve the hyperglycemia in diabetic mice. In the present study, we explored a novel protocol that reprogrammed primary hepatocytes into functional IPCs by using multicistronic vectors carrying pancreatic and duodenal homeobox-1 (Pdx1), neurogenin 3 (Ngn3), and v-musculoaponeurotic fibrosarcoma oncogene homolog A (MafA). These triple-transfected cells activated multiple beta-cell genes, synthesized and stored considerable amounts of insulin, and released the hormone in a glucose-regulated manner in vitro. Furthermore, when transplanted into streptozotocin-induced diabetic mice, the cells markedly ameliorated glucose tolerance. Our results indicated that ectopic expression of Pdx1, Ngn3, and MafA facilitated hepatocytes-to-IPCs reprogramming. This approach may offer opportunities for treatment of DM.

scholarly journals miR-11 regulates pupal size of Drosophila melanogaster via directly targeting Ras85D

2017 ◽  
Vol 312 (1) ◽  
pp. C71-C82 ◽  
Author(s):  
Yao Li ◽  
Shengjie Li ◽  
Ping Jin ◽  
Liming Chen ◽  
Fei Ma
Keyword(s):  
Drosophila Melanogaster ◽  
Untranslated Region ◽  
Deletion Mutant ◽  
Ectopic Expression ◽  
Whole Body ◽  
Physiological Processes ◽  
Insulin Producing Cells ◽  
Pupal Size

MicroRNAs play diverse roles in various physiological processes during Drosophila development. In the present study, we reported that miR-11 regulates pupal size during Drosophila metamorphosis via targeting Ras85D with the following evidences: pupal size was increased in the miR-11 deletion mutant; restoration of miR-11 in the miR-11 deletion mutant rescued the increased pupal size phenotype observed in the miR-11 deletion mutant; ectopic expression of miR-11 in brain insulin-producing cells (IPCs) and whole body shows consistent alteration of pupal size; Dilps and Ras85D expressions were negatively regulated by miR-11 in vivo; miR-11 targets Ras85D through directly binding to Ras85D 3′-untranslated region in vitro; removal of one copy of Ras85D in the miR-11 deletion mutant rescued the increased pupal size phenotype observed in the miR-11 deletion mutant. Thus, our current work provides a novel mechanism of pupal size determination by microRNAs during Drosophila melanogaster metamorphosis.

scholarly journals Ecdysterone receptor is a sequence-specific transcription factor involved in the developmental regulation of heat shock genes.

1991 ◽  
Vol 11 (7) ◽  
pp. 3660-3675 ◽  
Author(s):  
Y Luo ◽  
J Amin ◽  
R Voellmy
Keyword(s):  
Transcription Factor ◽  
Heat Shock ◽  
Binding Sites ◽  
Specific Binding ◽  
Developmental Regulation ◽  
Heat Shock Genes ◽  
Transfected Cells ◽  
Apparent Homogeneity ◽  
Transcription In Vitro

Purification of ecdysterone receptor from Drosophila melanogaster to apparent homogeneity is reported. Purified receptor binds specifically to several sequences in the promoters of the developmentally active hsp27 and hsp23 heat shock genes that were previously implied in ecdysterone regulation of the genes and that share limited homology among themselves and with mammalian steroid receptor binding sites. Some of these elements confer ecdysterone regulation on a basal promoter in transfected cells, acting in a synergistic fashion. Transcription in vitro of promoters containing such elements is stimulated up to 100-fold by added purified ecdysterone receptor, depending on receptor dosage and the number of elements present. Transcriptional enhancement requires sequence-specific binding of receptor to template promoters which facilitates the formation of a preinitiation complex. Ecdysterone stimulates DNA binding of the receptor in vitro.

scholarly journals eNOS deletion impairs mitochondrial quality control and exacerbates Western diet-induced NASH

2019 ◽  
Vol 317 (4) ◽  
pp. E605-E616 ◽  
Author(s):  
Ryan D. Sheldon ◽  
Grace M. Meers ◽  
E. Matthew Morris ◽  
Melissa A. Linden ◽  
Rory P. Cunningham ◽  
...  
Keyword(s):  
Quality Control ◽  
Transcription Factor ◽  
Mitochondrial Biogenesis ◽  
Western Diet ◽  
Acid Oxidation ◽  
Related Factor ◽  
Functional Impairments ◽  
Primary Hepatocytes ◽  
Mitochondrial Quality Control

Dysregulated mitochondrial quality control leads to mitochondrial functional impairments that are central to the development and progression of hepatic steatosis to nonalcoholic steatohepatitis (NASH). Here, we identify hepatocellular localized endothelial nitric oxide synthase (eNOS) as a novel master regulator of mitochondrial quality control. Mice lacking eNOS were more susceptible to Western diet-induced hepatic inflammation and fibrosis in conjunction with decreased markers of mitochondrial biogenesis and turnover. The hepatocyte-specific influence was verified via magnetic activated cell sorting purified primary hepatocytes and in vitro siRNA-induced knockdown of eNOS. Hepatic mitochondria from eNOS knockout mice revealed decreased markers of mitochondrial biogenesis (PPARγ coactivator-1α, mitochondrial transcription factor A) and autophagy/mitophagy [BCL-2-interacting protein-3 (BNIP3), 1A/1B light chain 3B (LC3)], suggesting decreased mitochondrial turnover rate. eNOS knockout in primary hepatocytes exhibited reduced fatty acid oxidation capacity and were unable to mount a normal BNIP3 response to a mitophagic challenge compared with wild-type mice. Finally, we demonstrate that eNOS is required in primary hepatocytes to induce activation of the stress-responsive transcription factor nuclear factor erythroid 2-related factor 2 ( NRF2). Thus, our data demonstrate that eNOS is an important regulator of hepatic mitochondrial content and function and NASH susceptibility.

Notch1 co-opts lymphoid enhancer factor 1 for survival of murine T-cell lymphomas

Blood ◽  
2007 ◽  
Vol 110 (7) ◽  
pp. 2650-2658 ◽  
Author(s):  
Christina Spaulding ◽  
Erica J. Reschly ◽  
Derek E. Zagort ◽  
Yumi Yashiro-Ohtani ◽  
Levi J. Beverly ◽  
...  
Keyword(s):  
Transcription Factor ◽  
T Cell ◽  
Cell Lymphoma ◽  
Ectopic Expression ◽  
Malignant Cell ◽  
T Cell Lymphomas ◽  
Notch1 Mutations ◽  
Enhancer Factor

Oncogenic Notch1 mutations are found in most T-lineage acute lymphoblastic leukemias in humans and T-cell lymphomas in mice. However, the mechanism by which Notch1 promotes transformation or maintains malignant cell survival has not been determined fully. Here, we report that expression of the transcription factor lymphoid enhancer factor 1 (Lef1) is Notch dependent in murine T-cell lymphomas in vitro and in vivo, and that the intracellular domain of Notch1 (ICN1) is present at the Lef1 promoter. Lef1 expression is not Notch dependent in primary T-cell progenitors, but Lef1 mRNA is increased by ectopic expression of ICN1 in these cells. We show that Lef1 is required for survival of T-cell lymphoma lines, and that ectopic expression of Lef1 delays lymphoma cell death in the absence of Notch signaling, indicating that Lef1 is an important Notch target in these cells. Therefore, Notch1 co-opts Lef1 during the process of transformation to maintain survival of T-cell lymphomas.

scholarly journals Evaluation of In Vivo Antidiabetic, In Vitro α-Amylase Inhibitory, and In Vitro Antioxidant Activity of Leaves Crude Extract and Solvent Fractions of Bersama abyssinica Fresen (Melianthaceae)

2020 ◽  
Vol 25 ◽  
pp. 2515690X2093582 ◽  
Author(s):  
Zemene Demelash Kifle ◽  
Engidaw Fentahun Enyew
Keyword(s):  
Diabetes Mellitus ◽  
Blood Glucose ◽  
Crude Extract ◽  
Oral Glucose ◽  
Diabetic Mice ◽  
Glucose Lowering ◽  
Treatment Of Diabetes ◽  
Glucose Lowering Effect ◽  
Bersama Abyssinica

Background. The leaves of Bersama abyssinica are used for the treatment of diabetes mellitus in folk medicine system of Ethiopia. The present study was done based on the traditional claim of B abyssinica for the treatment of diabetes mellitus. Methods. The α-amylase inhibition and antioxidant activities of B abyssinica extracts were evaluated by using 3,5-dinitrosalicylic acid method and diphenyl-2-picrylhydrazyl assay model, respectively. Blood glucose lowering activity of the extracts was studied in 4 animal models; normoglycemic, oral glucose loaded, and streptozotocin-induced diabetic mice models. Results. Among the extracts, the crude extract showed the highest α-amylase enzyme inhibition activity with an IC50 of 6.57 μg/mL. The water fraction showed the strongest antioxidant activity with an IC50 of 3.43 μg/mL. The crude extract at doses of 200, and 400 mg/kg showed significant ( P < .05) hypoglycemic activity in normoglycemic mice. All doses of the crude extract significantly ( P < .05) reduced blood glucose levels of oral glucose-loaded mice. In streptozotocin-induced diabetic mice models, both the crude and solvent fractions showed a significant ( P < .05) blood glucose lowering effect as compared with the negative control group post 8 hour treatment. Conclusion. The results demonstrated the beneficial biochemical effects of B abyssinica extract by inhibiting α-amylase and scavenging the free radicals. The crude extract and solvent fractions of B abyssinica had significant blood glucose lowering effect in all animal models.

Growth factor independent 1B (Gfi1b) is an E2A target gene that modulates Gata3 in T-cell lymphomas

Blood ◽  
2007 ◽  
Vol 109 (10) ◽  
pp. 4406-4414 ◽  
Author(s):  
Wei Xu ◽  
Barbara L. Kee
Keyword(s):  
Transcription Factor ◽  
Growth Factor ◽  
T Lymphocyte ◽  
Cell Expansion ◽  
Target Gene ◽  
Ectopic Expression ◽  
Growth Arrest ◽  
T Cell Lymphomas ◽  
Gata3 Expression

AbstractThe E2A transcription factors are required for normal T lymphopoiesis and to prevent T-lymphocyte progenitor transformation. Ectopic expression of E2A proteins in E2A-deficient lymphomas results in growth arrest and apoptosis, indicating that these cells remain responsive to the targets of E2A. Here we identify the transcriptional repressor growth factor independent 1B (Gfi1b) as a target of E2A that promotes growth arrest and apoptosis in lymphomas. Gfi1b expression in primary T-lymphocyte progenitors is dependent on E2A and excess Gfi1b prevents the outgrowth of T lymphocyte progenitors in vitro. Gfi1b represses expression of Gata3, a transcription factor whose appropriate regulation is required for survival of lymphomas and T-lymphocyte progenitors. We also show that ectopic expression of Gata3 in lymphomas promotes expression of Gfi1b, indicating that these proteins may function in an autoregulatory loop that maintains appropriate levels of Gata3. Therefore, we propose that E2A proteins prevent lymphoma cell expansion, at least in part through regulation of Gfi1b and modulation of Gata3 expression.

Ecdysterone receptor is a sequence-specific transcription factor involved in the developmental regulation of heat shock genes

1991 ◽  
Vol 11 (7) ◽  
pp. 3660-3675
Author(s):  
Y Luo ◽  
J Amin ◽  
R Voellmy
Keyword(s):  
Transcription Factor ◽  
Heat Shock ◽  
Binding Sites ◽  
Specific Binding ◽  
Developmental Regulation ◽  
Heat Shock Genes ◽  
Transfected Cells ◽  
Apparent Homogeneity ◽  
Transcription In Vitro

Purification of ecdysterone receptor from Drosophila melanogaster to apparent homogeneity is reported. Purified receptor binds specifically to several sequences in the promoters of the developmentally active hsp27 and hsp23 heat shock genes that were previously implied in ecdysterone regulation of the genes and that share limited homology among themselves and with mammalian steroid receptor binding sites. Some of these elements confer ecdysterone regulation on a basal promoter in transfected cells, acting in a synergistic fashion. Transcription in vitro of promoters containing such elements is stimulated up to 100-fold by added purified ecdysterone receptor, depending on receptor dosage and the number of elements present. Transcriptional enhancement requires sequence-specific binding of receptor to template promoters which facilitates the formation of a preinitiation complex. Ecdysterone stimulates DNA binding of the receptor in vitro.

In vitro generation of HSC-like cells from murine ESCs/iPSCs by enforced expression of LIM-homeobox transcription factor Lhx2

Blood ◽  
2011 ◽  
Vol 117 (14) ◽  
pp. 3748-3758 ◽  
Author(s):  
Kenji Kitajima ◽  
Ken-ichi Minehata ◽  
Kenji Sakimura ◽  
Toru Nakano ◽  
Takahiko Hara
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Ectopic Expression ◽  
Hematopoietic Cells ◽  
Embryonic Stem ◽  
Hematopoietic Stem ◽  
Homeobox Transcription Factor ◽  
Induced Pluripotent

Abstract Identification of genes involved in in vitro differentiation induction of embryonic stem cells (ESCs) into hematopoietic stem cells (HSCs) has been challenged during last decade. To date, a homeobox transcription factor Hoxb4 has been only demonstrated to possess such an effect in mice. Here, we show that HSC-like cells were efficiently induced from mouse ESCs by enforced expression of Lhx2, a LIM-homeobox transcription factor. Transduction of Lhx2 into ESC-derived mesodermal cells resulted in robust differentiation of c-Kit+/Sca-1+/Lineage− (KSL) cells in vitro. The KSL cell induction frequency was superior to the case of Hoxb4. Furthermore, transplantation of Lhx2-transduced hematopoietic cells into lethally irradiated mice resulted in multilineage repopulation of hematopoietic cells over 4 months. Transduction of Lhx2 into induced pluripotent stem cells (iPSCs) was also effective in generating KSL cells in vitro, as well as HSC-like activities in vivo. These results demonstrate that ectopic expression of Lhx2 confers an in vivo engrafting capacity to ESC/iPSC-derived hematopoietic cells and in vivo behavior of iPSC-derived hematopoietic cells is almost identical to that of ESC-derived cells.

scholarly journals High Norepinephrine Status Induced Growth of Colorectal Cancer With Type 2 Diabetes Mellitus Benefited From ADP-Ribosyltransferase 1

2020 ◽  
Author(s):  
Wenwen Chen ◽  
Yanping Yang ◽  
Jiale Duan ◽  
Ming Li ◽  
Yi Tang ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Colorectal Cancer ◽  
Type 2 Diabetes ◽  
Type 2 Diabetes Mellitus ◽  
Diabetic Mice ◽  
Transplanted Tumors ◽  
Norepinephrine Level ◽  
Adp Ribosyltransferase

Abstract BackgroundIt is known that type 2 diabetes mellitus patients have higher susceptibility to colorectal cancer and poorer prognosis, but the mechanism is quite unknown. Here, we investigated effect of ADP-Ribosyltransferase 1 on growth of colorectal cancer combined diabetes in high norepinephrine status and the potential mechanism. MethodsWe evaluated size and weight of transplanted tumors with different ADP-Ribosyltransferase 1 level of CT26 cells or different norepinephrine level on diabetic mice model and observed their survival time as well. Consistently, CCK8 and flow cytometry were applied for detecting growth of CT26 cells in vitro. Western blot was performed for analyzing differentially expressed proteins of proliferatic profiles to determine ADP-Ribosyltransferase 1-modulated pathway.ResultsAccording to our data, high level of norepinephrine and ADP-Ribosyltransferase 1 both facilitated proliferation of CT26 cells in vitro and in vivo, besides, inhibition of norepinephrinee-depended-proliferation was observed in ADP-Ribosyltransferase 1 silencing CT26 cells in vitro compared with CT26 cells with ADP-Ribosyltransferase 1 expression. However, we discovered after reducing norepinephrine level of serum by surgery, size and weight of the transplanted tumors were significantly reduced compared with non-operated group and sham-operated group. Further, expression of ADP-Ribosyltransferase 1, mTOR, STAT3, p-AKT protein in tumor tissues of diabetic mice was increased rather than non-diabetes mice, while after depleting norepinephrine level by renal denervation operation, expression of proliferation-relative proteins mTOR, STAT3, p-AKT protein was decreased, but no change was discovered in ADP-Ribosyltransferase 1 expression. While under the same concentration of norepinephrine environment, ADP-Ribosyltransferase 1 boosts expression of p-AKT, mTOR, STAT3, CyclinD1 and c-myc in CT26 cells in vitro. ConclusionsThis study proposed a hypothesis that high-norepinephrine-induced proliferation of colorectal cancer required expression of ADP-Ribosyltransferase 1, and raise ADP-Ribosyltransferase 1 might be a candidate target for treatment of diabetes-associated colorectal cancer.

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