Cardio Vascular Disease, Diabete Type 2
Conditions
Brief summary
This study aims to isolate endothelial progenitor cells (EPCs) from participants with type 2 diabetes (T2D) and cardiovascular complications and to comprehensively characterize EPC dysfunction. Specifically, the study will evaluate maladaptive angiocrine signaling, calcium signaling pathways, and the role of inflammation in EPC function and the progression of atherosclerosis during T2D development. A sub-study will assess EPC functionality by examining endothelial nitric oxide synthase (eNOS) expression and activity, as well as the effectiveness of in vitro eNOS gene enhancement.
Detailed description
Type 2 diabetes (T2D) is associated with damage to blood vessels, which can lead to serious complications such as heart disease, stroke, and other vascular problems. Endothelial progenitor cells (EPCs) help maintain healthy blood vessels by repairing vascular injury. In people with T2D, the number and function of these cells are reduced, which may contribute to poor blood vessel repair and increased cardiovascular risk. The mechanisms responsible for this dysfunction are not fully understood. This study aims to examine how changes in cell signaling and chronic low-grade inflammation in T2D affect EPC function. EPCs will be isolated from patients with T2D, with and without cardiovascular complications, to assess their signaling properties, function, and ability to mature into vascular cells. An in vitro sub-study will evaluate a potential therapeutic strategy to improve EPC function by increasing the activity of endothelial nitric oxide synthase (eNOS), a protein that plays a key role in maintaining healthy blood vessels. Reduced eNOS activity is an important contributor to vascular dysfunction in diabetes. Enhancing eNOS expression and function in EPCs may improve their regenerative capacity and help prevent or treat diabetic vascular complications.
Interventions
None listed
Sponsors
Weill Cornell Medical College in Qatar
Hamad Medical Corporation
Study design
Observational model
COHORT
Time perspective
PROSPECTIVE
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
Yes
Inclusion criteria
* T2D * Males and females * Older than 18 years of age * Willingness to participate in the study and provide written consent form * Consent to having peripheral blood withdrawals and urine collection for the study requirement.
Exclusion criteria
* Unable to meet the inclusion criteria * Type I diabetes, MODY diabetes or other form of diabetes * Active infection, inflammation, cancer or acute illness of any kind (other than a cardiovascular complication of diabetes if applicable in the group they are assigned to). * Chronic inflammation (eg. auto-immune diseases) or infections (eg. HIV, chronic hepatitis). * Evidence of malignancy within the past 5 years
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Characterization of EPCs dysregulation in Type 2 diabetes | Following the EPC isolation (15 - 20 days). | Functional characterization of endothelial progenitor cells (EPCs) isolated from participants will be performed by assessing cellular signaling and functional pathways. Measurements will be compared among EPCs isolated from participants with type 2 diabetes and cardiovascular diseases, participants with type 2 diabetes without cardiovascular diseases, and healthy volunteers without diabetes or cardiovascular diseases. |
| Functional Analysis of inflammatory responses in Endothelial Progenitor Cells in type 2 Diabetes with cardiovascular complications | Following the EPC isolation (15 - 20 days) | Functional characterization of endothelial progenitor cells (EPCs) isolated from participants will be performed by assessing cellular signaling and functional pathways. Measurements will be compared among EPCs isolated from participants with type 2 diabetes and cardiovascular diseases, participants with type 2 diabetes without cardiovascular diseases, and healthy volunteers without diabetes or cardiovascular diseases. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Expression levels of angiocrine factor genes in EPCs | Following the EPC isolation (15-20 days) | Expression levels of angiocrine related genes in EPCs measured by RNA sequencing and reported as normalized transcript counts. |
| Cytosolic calcium concetration in EPCs | Following the EPC isolation (15-20 days) | Cytosolic calcium concentration and expression of calcium signaling transcripts and proteins in EPCs assessed using functional calcium assays and molecular analyses. |
| Quantification of Mitochondrial reactive oxygen species (ROS) levels in EPCs | Following the EPC isolation (15-20 days). | Mitochondrial ROS production in EPCs measured using pre-designed fluorescence-based assay kits and reported as relative fluorescence units. Comparisons will be made among the three study groups. |
| Protein Expression of Inflammatory Transcription Factors in Endothelial Progenitor Cells | Following the EPC isolation (15 - 20 days). | Protein expression levels in lysates are quantified using western blot analysis and normalized to housekeeping proteins. |
| Angiogenic Transcription Factor Expression in EPCs | Time Frame: Following the EPC isolation (15 - 20 days). | Protein expression levels in lysates are quantified using western blot analysis and normalized to housekeeping proteins. |
| Proportion of EPCs differentiating into endothelial cells in vitro | 14 days after initiation of in vitro differentiation. | Differentiation of EPCs into adherent endothelial cells following culture in VEGF-, FGF-2-, IGF-1-, and heparin-supplemented media, assessed by endothelial morphology and marker expression. |
| Measurement of eNOS(endothelial nitric oxide synthase)-Dependent Nitric Oxide Production in Endothelial Progenitor Cells | Following EPC isolation (15-20 days) and 72 hours post-gene transfection. | eNOS(endothelial nitric oxide synthase) mRNA expression measured by RT-PCR and eNOS protein expression measured by western blotting in EPCs isolated from participants with type 2 diabetes and healthy volunteers. |
| Functional Outcomes of Genetically Enhanced Endothelial Progenitor Cells | Following EPC isolation (15-20 days) and 72 hours post-gene transfection | eNOS mRNA expression measured by RT-PCR and eNOS protein expression measured by western blotting in EPCs isolated from participants with type 2 diabetes and healthy volunteers. |
Countries
Qatar
Contacts
PRINCIPAL_INVESTIGATORCharbel Abi Khalil, MD,PhD
Weill Cornell Medicine-Qatar
Outcome results
None listed