scholarly journals Modulation of Insulin Gene Expression with CRISPR/Cas9-based Transcription Factors

2021 ◽  
Vol 9 (A) ◽  
pp. 876-881
Author(s):  
Bakhytzhan Alzhanuly ◽  
Zhussipbek Y. Mukhatayev ◽  
Dauren M. Botbayev ◽  
Yeldar Ashirbekov ◽  
Nurlybek D. Katkenov ◽  
...  
Keyword(s):  
Embryonic Stem ◽  
Cell Types ◽  
Diabetic Patients ◽  
Type I ◽  
Transcription Activator ◽  
New Approach ◽  
Donor Cells ◽  
Diabetes Type I ◽  
Promising Solution ◽  
Insulin Gene Expression

Background: The discovery and use of CRISPR/Cas9 technology have enabled researchers throughout the globe to continuously edit genomes for the benefit of science and medicine. Diabetes type I is one field of medicine where CRISPR/Cas9 has a strong potential for cell therapy development. The long-lasting paucity of healthy cells for clinical transplantation into diabetic patients has led to the search of new methods for producing β-cells from other human cell types. Embryonic stem cells are being studied worldwide as one most promising solution of this need. Aim: The aim of the study is to to check the feasibility of modulating human insulin transcription using CRISPR/Cas9-based synthetic transcription regulation factors. Results: A new approach for creating potential therapeutic donor cells with enhanced and suppressed insulin production based on one of the latest achievements of human genome editing was developed. Both synthetic transcription activator (VP64) and transcription repressor (KRAB) proteins were shown to function adequately well as a part of the whole CRISPR/Cas9-based system. We claim that our results have a lot to offer and can bring light to many studies where numerous labs are struggling on to treat this disease.

Stem Cell-Based Therapies: A New Ray of Hope for Diabetic Patients

2019 ◽  
Vol 14 (2) ◽  
pp. 146-151 ◽  
Author(s):  
Junaid Khan ◽  
Amit Alexander ◽  
Mukta Agrawal ◽  
Ajazuddin ◽  
Sunil Kumar Dubey ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Drug Therapy ◽  
Type I Diabetes ◽  
Embryonic Stem ◽  
Health Concern ◽  
Future Research ◽  
Diabetic Patients ◽  
Successful Implementation ◽  
Type I

Diabetes and its complications are a significant health concern throughout the globe. There are physiological differences in the mechanism of type-I and type-II diabetes and the conventional drug therapy as well as insulin administration seem to be insufficient to address the problem at large successfully. Hypoglycemic swings, frequent dose adjustments and resistance to the drug are major problems associated with drug therapy. Cellular approaches through stem cell based therapeutic interventions offer a promising solution to the problem. The need for pancreatic transplants in case of Type- I diabetes can also be by-passed/reduced due to the formation of insulin producing β cells via stem cells. Embryonic Stem Cells (ESCs) and induced Pluripotent Stem Cells (iPSCs), successfully used for generating insulin producing β cells. Although many experiments have shown promising results with stem cells in vitro, their clinical testing still needs more exploration. The review attempts to bring into light the clinical studies favoring the transplantation of stem cells in diabetic patients with an objective of improving insulin secretion and improving degeneration of different tissues in response to diabetes. It also focuses on the problems associated with successful implementation of the technique and possible directions for future research.

scholarly journals Bilateral Diabetic Knee Neuroarthropathy in a Forty-Year-Old Patient

2016 ◽  
Vol 2016 ◽  
pp. 1-4 ◽  
Author(s):  
Patrick Goetti ◽  
Nicolas Gallusser ◽  
Olivier Borens
Keyword(s):  
Joint Destruction ◽  
Young Patients ◽  
Joint Disease ◽  
Diabetic Patients ◽  
Type I ◽  
Bilateral Involvement ◽  
Satisfactory Outcome ◽  
History Of ◽  
Diabetes Type I ◽  
Low Incidence

Diabetic osteoarthropathy is a rare cause of neuropathic joint disease of the knee; bilateral involvement is even more exceptional. Diagnosis is often made late due to its unspecific symptoms and appropriate surgical management still needs to be defined, due to lack of evidence because of the disease’s low incidence. We report the case of a forty-year-old woman with history of diabetes type I who developed bilateral destructive Charcot knee arthropathy. Bilateral total knee arthroplasty was performed in order to achieve maximal functional outcome. Follow-up was marked by bilateral tibial periprosthetic fractures treated by osteosynthesis with a satisfactory outcome. The diagnosis of Charcot arthropathy should always be in mind when dealing with atraumatic joint destruction in diabetic patients. Arthroplasty should be considered as an alternative to arthrodesis in bilateral involvement in young patients.

scholarly journals Management of Diabetic Ketoacidosis in Patients with Diabetes Type I

2019 ◽  
Author(s):  
Basam Zuhaer Sindi ◽  
Naif Mishkhas Alazwari ◽  
Abdulaziz Mohammed Khateeb ◽  
Aqil Abdulmonem Alzaher ◽  
Mohammad Abdulaziz M Alkhawajah ◽  
...  
Keyword(s):  
Diabetic Ketoacidosis ◽  
Fluid Replacement ◽  
Diabetic Patients ◽  
Type I ◽  
Care Providers ◽  
Patients With Diabetes ◽  
Type One Diabetes ◽  
Diabetes Type I ◽  
Comprehensive Search ◽  
Health Complications

Background: DKA is the leading cause of mortality among pediatric age and young adults with T1D, responsible for almost 50% of all fatalities in diabetic patients younger than 24 years of age. Goals of DKA management include optimization of volume status, hyperglycemia and ketoacidosis, electrolyte abnormalities, and potential precipitating factors. Methodology: we conducted this review using a comprehensive search of MEDLINE, PubMed, and EMBASE, January 1987, through February 2017. The following search terms were used: ketoacidosis, management of diabetic ketoacidosis, type I diabetic patients’ emergency complication, fluid replacement in DKA, insulin therapy approach. Aim: in this review, we aim at evaluating the various ways of approaching patients who suffer from type-one diabetes during ketoacidosis and keto-acidotic coma. We will try to understand the triggers and pathophysiology behind this condition, and explore ways to prevent them. Conclusion: prompt diagnoses, aggressive treatment, and education of patient and their care providers about prevention strategies must be implemented. Also, more studies are required in the area of preventing health complications due to these types of diseases to effectively manage DKA in the future.

scholarly journals MicroRNA-deficient mouse embryonic stem cells acquire a functional interferon response

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Jeroen Witteveldt ◽  
Lisanne I Knol ◽  
Sara Macias
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Mammalian Cells ◽  
Viral Infections ◽  
Embryonic Stem ◽  
Cell Types ◽  
Interferon Response ◽  
Central Component ◽  
Type I ◽  
Antiviral Mechanism

When mammalian cells detect a viral infection, they initiate a type I interferon (IFNs) response as part of their innate immune system. This antiviral mechanism is conserved in virtually all cell types, except for embryonic stem cells (ESCs) and oocytes which are intrinsically incapable of producing IFNs. Despite the importance of the IFN response to fight viral infections, the mechanisms regulating this pathway during pluripotency are still unknown. Here we show that, in the absence of miRNAs, ESCs acquire an active IFN response. Proteomic analysis identified MAVS, a central component of the IFN pathway, to be actively silenced by miRNAs and responsible for suppressing IFN expression in ESCs. Furthermore, we show that knocking out a single miRNA, miR-673, restores the antiviral response in ESCs through MAVS regulation. Our findings suggest that the interaction between miR-673 and MAVS acts as a switch to suppress the antiviral IFN during pluripotency and present genetic approaches to enhance their antiviral immunity.

36 RESTORATION OF TELOMERE LENGTH IN CLONED PIG EMBRYOS DURING EARLY EMBRYOGENESIS IS NOT DEPENDENT ON TELOMERE LENGTH AND TYPE OF DONOR CELLS

2013 ◽  
Vol 25 (1) ◽  
pp. 166
Author(s):  
T. Q. Dang-Nguyen ◽  
S. Haraguchi ◽  
S. Akagi ◽  
T. Somfai ◽  
M. Kaneda ◽  
...  
Keyword(s):  
Telomere Length ◽  
Embryonic Stem ◽  
Donor Cell ◽  
Cell Types ◽  
Telomere Elongation ◽  
Dependent Process ◽  
Donor Cells ◽  
Different Types ◽  
Cloned Pig

Analyses on telomere length in cloned animals have revealed diverse results depending on the donor cell types. In mice and cattle, telomere length is reset during morula-blastocyst transition and the restoration is thought to be a telomerase-dependent process. However, it is still unknown whether the pattern of telomere elongation during this transition is dependent on donor cell types. In the present study, we examined the changes of telomere length during morula-blastocyst transition in cloned porcine embryos using different types of donor cell. Embryonic stem-like cells (ES), cumulus cells (C), fibroblasts at passages 7 and 10 (F7 and F10, respectively) were used as donor cells to produce NT embryos (ES, C, F7, and F10 groups, respectively). Telomere lengths of ES (35.8 ± 1.5 kb), C (24.4 ± 0.5 kb), P7 (18.7 ± 0.6 kb), and P10 (17.2 ± 0.1 kb) cells were significantly different. In contrast, cloned morulae in ES, C, F7, and F10 groups did not have any significant differences in telomere length (18.2 ± 0.3, 17.8 ± 0.7, 18.5 ± 0.3, and 18.4 ± 0.4 kb, respectively). Likewise, cloned blastocysts in ES, C, F7, and F10 groups had similar telomere length (22.3 ± 1.5, 23.5 ± 2.6, 20.2 ± 1.0, and 20.9 ± 1.0 kb, respectively). However, the telomere of the blastocyst was significantly longer (P < 0.05) compared with the morula in the respective group. Furthermore, relative telomerase activities of cloned morulae in ES, C, F7, and F10 groups (4.2 ± 0.4, 4.0 ± 0.5, 5.1 ± 0.4, and 4.9 ± 0.4, respectively) were significantly lower (P < 0.01) than those of cloned blastocysts in the same groups (8.2 ± 1.1, 8.6 ± 0.6, 12.5 ± 2.9, and 8.3 ± 1.1, respectively). The proportions of blastocysts in cloned embryos for ES, C, F7, and F10 groups (10.0 ± 5.2, 17.3 ± 2.9, 13.5 ± 2.9, and 13.1 ± 3.6%, respectively) did not significantly differ. Total cell numbers in blastocysts for ES, C, F7, and F10 groups (28.3 ± 2.9, 32.6 ± 3.6, 30.4 ± 3.1, and 27.4 ± 2.2, respectively) were not significantly different as well. In the present study, we found that the telomere elongation in cloned pig embryos occurs during morula-blastocyst transition. This is consistent with the previous findings in in vivo and in vitro fertilization and cloned embryos in cattle and mice. We also revealed that although different types of cells (ES, C, and F) or the same type of cells with different telomere length (F7 and F10) were used for NT, their resultant morulae and blastocysts had similar telomere length. This suggests that the telomere restoration during morula-blastocyst transition is independent of telomere length and type of donor cells. An increase in telomerase activity during morula-blastocyst transition indicates that the elongation of telomere length was likely a telomerase-dependent process. In conclusion, restoration of telomere length in cloned porcine embryos during morula-blastocyst transition was independent of telomere length and type of donor cells, and likely a telomerase-dependent process.

scholarly journals Hydrogel platform for in vitro three-dimensional assembly of human stem cell-derived β cells and endothelial cells

2019 ◽  
Author(s):  
Punn Augsornworawat ◽  
Leonardo Velazco-Cruz ◽  
Jiwon Song ◽  
Jeffrey R. Millman
Keyword(s):  
Endothelial Cells ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Cell Types ◽  
Diabetic Patients ◽  
Β Cells

AbstractDifferentiation of stem cells into functional replacement cells and tissues is a major goal of the regenerative medicine field. However, one limitation has been organization of differentiated cells into multi-cellular, three-dimensional assemblies. The islets of Langerhans contain many endocrine and non-endocrine cell types, such as insulin-producing β cells and endothelial cells. Transplantation of exogenous islets into diabetic patients can serve as a cell replacement therapy, replacing the need for patients to inject themselves with insulin, but the number of available islets from cadaveric donors is low. We have developed a strategy of assembling human embryonic stem cell-derived β cells with endothelial cells into three-dimensional aggregates on a hydrogel. The resulting islet organoids express β cell markers and are functional, capable of undergoing glucose-stimulated insulin secretion. These results provide a platform for evaluating the effects of the islet tissue microenvironment on human embryonic stem cell-derived β cells and other islet endocrine cells to develop tissue engineered islets.

scholarly journals Management of Diabetic Ketoacidosis in Patients with Diabetes Type I

2019 ◽  
Author(s):  
Basam Zuhaer Sindi ◽  
Naif Mishkhas Alazwari ◽  
Abdulaziz Mohammed Khateeb ◽  
Aqil Abdulmonem Alzaher ◽  
Mohammad Abdulaziz M Alkhawajah ◽  
...  
Keyword(s):  
Diabetic Ketoacidosis ◽  
Fluid Replacement ◽  
Diabetic Patients ◽  
Type I ◽  
Care Providers ◽  
Patients With Diabetes ◽  
Type One Diabetes ◽  
Diabetes Type I ◽  
Comprehensive Search ◽  
Health Complications

Background: DKA is the leading cause of mortality among pediatric age and young adults with T1D, responsible for almost 50% of all fatalities in diabetic patients younger than 24 years of age. Goals of DKA management include optimization of volume status, hyperglycemia and ketoacidosis, electrolyte abnormalities, and potential precipitating factors. Methodology: we conducted this review using a comprehensive search of MEDLINE, PubMed, and EMBASE, January 1987, through February 2017. The following search terms were used: ketoacidosis, management of diabetic ketoacidosis, type I diabetic patients’ emergency complication, fluid replacement in DKA, insulin therapy approach. Aim: in this review, we aim at evaluating the various ways of approaching patients who suffer from type-one diabetes during ketoacidosis and keto-acidotic coma. We will try to understand the triggers and pathophysiology behind this condition, and explore ways to prevent them. Conclusion: prompt diagnoses, aggressive treatment, and education of patient and their care providers about prevention strategies must be implemented. Also, more studies are required in the area of preventing health complications due to these types of diseases to effectively manage DKA in the future.

scholarly journals MicroRNA-deficient embryonic stem cells acquire a functional Interferon response

2018 ◽  
Author(s):  
Jeroen Witteveldt ◽  
Lisanne Knol ◽  
Sara Macias
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Mammalian Cells ◽  
Viral Infections ◽  
Embryonic Stem ◽  
Cell Types ◽  
Interferon Response ◽  
Central Component ◽  
Type I ◽  
Antiviral Mechanism

When mammalian cells detect a viral infection, they initiate a type-I Interferon (IFNs) response as part of their innate immune system. This antiviral mechanism is conserved in virtually all cell types, except for embryonic stem cells (ESCs) and oocytes which are intrinsically incapable of producing IFNs. Despite the importance of the IFN response to fight viral infections, the mechanisms regulating this pathway during pluripotency are still unknown. Here we show that, in the absence of miRNAs, ESCs acquire an active IFN response. Proteomic analysis identified MAVS, a central component of the IFN pathway, to be actively silenced by miRNAs and responsible for suppressing IFN expression in ESCs. Furthermore, we show that knocking out a single miRNA, miR-673, restores the antiviral response in ESCs through MAVS regulation. Our findings suggest that the interaction between miR-673 and MAVS acts as a switch to suppress the antiviral IFN during pluripotency and present genetic approaches to enhance their antiviral immunity.

Processing Immunoperoxidase Labeled Cell Monolayers and Cryostat Sesctions For Electron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 714-715
Author(s):  
K. Chien ◽  
R. Van de Velde ◽  
I.P. Shintaku ◽  
A.F. Sassoon
Keyword(s):  
Cell Monolayer ◽  
Cell Types ◽  
Surface Antigens ◽  
Immunoperoxidase Method ◽  
Reaction Step ◽  
Hot Plate ◽  
Air Drying ◽  
New Approach ◽  
Cell Monolayers ◽  
Glass Slides

Immunoelectron microscopy of neoplastic lymphoma cells is valuable for precise localization of surface antigens and identification of cell types. We have developed a new approach in which the immunohistochemical staining can be evaluated prior to embedding for EM and desired area subsequently selected for ultrathin sectioning.A freshly prepared lymphoma cell suspension is spun onto polylysine hydrobromide- coated glass slides by cytocentrifugation and immediately fixed without air drying in polylysine paraformaldehyde (PLP) fixative. After rinsing in PBS, slides are stained by a 3-step immunoperoxidase method. Cell monolayer is then fixed in buffered 3% glutaraldehyde prior to DAB reaction. After the DAB reaction step, wet monolayers can be examined under LM for presence of brown reaction product and selected monolayers then processed by routine methods for EM and embedded with the Chien Re-embedding Mold. After the polymerization, the epoxy blocks are easily separated from the glass slides by heatingon a 100°C hot plate for 20 seconds.

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