From Skin Fibroblasts to Neural Stem Cells to Investigate in Vitro the Impact of Diabetes on Adult Neurogenesis

Active, not recruiting

• Conditions: Cognitive Dysfunction; Diabetes Mellitus Type 2 in Obese; Metabolism Disorder; Diabetes Mellitus, Type 2; Diabetes Mellitus Type 2 in Nonobese; Obesity
• Interventions: Other: Skin biopsy

• Registration Number: NCT05755321
• Lead Sponsor: Fondazione Policlinico Universitario Agostino Gemelli IRCCS

Brief Summary

Obesity and glucose intolerance or overt diabetes are increasing at an alarming rate in the population, and are bound to become a public health issue and a major cause of disability, loss of independence and high social costs in the near future. A large body of evidence has in recent years highlighted, among the negative effects of overnutrition and glucose dysmetabolism, also an acceleration of cognitive decline and of brain senescence, through cellular (vascular, neuronal, or both) and molecular mechanisms still incompletely clarified. Understanding how overweight and impaired glucose homeostasis negatively affect brain function represents both a major scientific challenge and an avenue to early detection and possibly prevention of this invalidating complication. The aim of this project is to obtain neuronal progenitor-like cells from skin fibroblasts in order to correlate patient-specific metabolism to adult neural stem cell (NSC) and neuronal function in vitro.

Detailed Description

* Analysis of skin fibroblasts from normal, obese, lean/diabetic and obese/diabetic individuals and in vitro epigenetic conversion, as revealed by morphological changes, loss of mesenchymal markers and expression of pluripotency genes.

* To re-differentiate patient-derived pluripotent cells into neural progenitor (NP) and to test their proliferative, self-renewal and multilineage differentiation capacity.

* To evaluate neuronal cells derived from patient and control pluripotent cells for a number functional and biochemical parameters (apoptosis/autophagy, mitochondrial potential, reactive oxygen species etc.) relevant to neurodegenerative disorders.

Study Timeline

• Study Start: 2020-01-01
• Status Verified: 2023-02
• Study First Submitted: 2023-02-23
• Study First Submitted QC: 2023-02-23
• Study First Posted: 2023-03-06
• Last Update Submitted: 2023-03-08
• Last Update Posted: 2023-03-10
• Primary Completion: 2023-04-30
• Study Completion: 2023-05-14

Recruitment & Eligibility

• Status: ACTIVE_NOT_RECRUITING
• Sex: All
• Target Recruitment: 40

Inclusion Criteria

• Patients of both sexes with BMI < 25 (normal weight) and BMI > 30 (obese) with or without T2DM with an indication for surgery

Exclusion Criteria

• Malignant neoplastic diseases

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Arm && Interventions

Group	Intervention	Description
BMI<25 without T2DM	Skin biopsy	10 Subjects with Body Mass Index (BMI) \<25 without Type 2 Diabetes Mellitus (T2DM) of both sex.
BMI>30 and T2DM	Skin biopsy	10 Subjects with Body Mass Index (BMI) \>30 with Type 2 Diabetes Mellitus (T2DM) of both sex.
BMI>30 without T2DM	Skin biopsy	10 Subjects with Body Mass Index (BMI) \>30 without Type 2 Diabetes Mellitus (T2DM) of both sex.
BMI<25 and T2DM	Skin biopsy	10 Subjects with Body Mass Index (BMI) \<25 with Type 2 Diabetes Mellitus (T2DM) of both sex.

Primary Outcome Measures

Name	Time	Method
Characterization of neural progenitors cells	At the end of the epigenetic conversion, the cell cultures are "blocked" and analysed with biochemistry and molecular biology techniques whose protocols and data analysis require 30 days not necessarily continuous	To determine whether neural progenitor cell (NPCs) obtained from dysmetabolic individuals are defective (analysis of degenerative features) when compared with control-derived cells.
Fibroblasts culture characterization	About one month to obtain cell cultures from each biopsy. Approximately one month (not necessarily continuous) for in vitro experiments and analyses. Total: 60 days per sample	To determine whether cells from dysmetabolic individuals are defective (in term of proliferation, senescence, cell death) when compared with control-derived cells
Efficiency of fibroblast epigenetic conversion (gene expression analysis)	The epigenetic conversion protocol requires 25 days, the analysis of gene expression (qRTPCR) approximately 5 days.	To determine whether cells from dysmetabolic individuals are different in term of epigenetic conversion efficiency when compared with control-derived cells.

Trial Locations

Locations (1)

Mingrone Geltrude; 🇮🇹 Roma, Italy