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

Study Overview
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Brief Summary
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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
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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.

Official Title
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From Skin to Brain: Epigenetic Conversion of Skin Fibroblasts Into Neural Stem Cells to Investigate the Impact of Diabetes on Adult Neurogenesis and Neuronal Function

Conditions
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Diabetes Mellitus Type 2 in Nonobese, Obesity, Cognitive Dysfunction, Metabolism Disorder, Diabetes Mellitus, Type 2, Diabetes Mellitus Type 2 in Obese

Intervention / Treatment
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* Other: Skin biopsy


Participation Criteria
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Eligibility Criteria
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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
Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| BMI<25 and T2DM<br>10 Subjects with Body Mass Index (BMI) <25 with Type 2 Diabetes Mellitus (T2DM) of both sex. | Other: Skin biopsy<br>* Skin biopsy performed during surgery.<br>|
| BMI<25 without T2DM<br>10 Subjects with Body Mass Index (BMI) <25 without Type 2 Diabetes Mellitus (T2DM) of both sex. | Other: Skin biopsy<br>* Skin biopsy performed during surgery.<br>|
| BMI>30 and T2DM<br>10 Subjects with Body Mass Index (BMI) >30 with Type 2 Diabetes Mellitus (T2DM) of both sex. | Other: Skin biopsy<br>* Skin biopsy performed during surgery.<br>|
| BMI>30 without T2DM<br>10 Subjects with Body Mass Index (BMI) >30 without Type 2 Diabetes Mellitus (T2DM) of both sex. | Other: Skin biopsy<br>* Skin biopsy performed during surgery.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Fibroblasts culture characterization | To determine whether cells from dysmetabolic individuals are defective (in term of proliferation, senescence, cell death) when compared with control-derived cells | 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 | 
| Efficiency of fibroblast epigenetic conversion (gene expression analysis) | To determine whether cells from dysmetabolic individuals are different in term of epigenetic conversion efficiency when compared with control-derived cells. | The epigenetic conversion protocol requires 25 days, the analysis of gene expression (qRTPCR) approximately 5 days. | 
| Characterization of neural progenitors cells | To determine whether neural progenitor cell (NPCs) obtained from dysmetabolic individuals are defective (analysis of degenerative features) when compared with control-derived 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 |