Based on solid preclinical results in mice and preliminary data in humans, this study aims to provide the proof of concept of the crucial role of the hematopoietic process occurring in human adipose tissue in the initiation of the inflammatory process at the origin of insulin resistance and type 2 diabetes (T2D). The main objective is to compare the number of pro-inflammatory macrophages derived from human adipose tissue hematopoietic stem cells (HSC) according to their origin, type 2 diabetes subjects or healthy volunteers.
In mice, increasing data demonstrate a causal relationship between the inflammatory process in adipose tissue and the development of insulin resistance, resulting in type 2 diabetes occurrence. However, the mechanisms mediating inflammation and its metabolic consequences are still unclear. In humans, recent publications have suggested an important interaction between the metabolic status and medullar hematopoietic activity. A preliminary study performed by the STROMAlab's research team has identified functional hematopoietic stem cells in human adipose tissue samples, indicating that adipose tissue-hematopoiesis is an active mechanism. To determine whether an alteration in adipose tissue-hematopoiesis could be a hallmark of type 2 diabetes in human, biopsies of subcutaneous adipose tissue will be performed in 2 groups of 10 volunteers: overweight/obese type 2 diabetes subjects versus healthy volunteers, matched on age. Then, hematopoietic stem cells extracted from human biopsies will be grafted into immunodeficient mice and after 12 weeks, flow cytometry using antibodies specific for human cell surface markers will be performed to quantify proinflammatory macrophages derived from human adipose tissue-hematopoietic stem cells.
Study Type
INTERVENTIONAL
Allocation
NA
Purpose
BASIC_SCIENCE
Masking
NONE
Enrollment
20
Abdominal subcutaneous biopsies and blood test for each volunteer.
CHU de Toulouse - Rangueil
Toulouse, France
Number of pro-inflammatory macrophages derived from human adipose tissue in mice transplanted with adipose tissue-hematopoietic stem cells .
The investigators expect that the number of pro-inflammatory macrophages derived from human adipose tissue will be significantly higher in mice transplanted with adipose tissue-hematopoietic stem cells isolated from diabetic subjects compared to those from healthy volunteers.
Time frame: Day 3 - Day 45
Comparison of hematopoietic activity in vitro between both groups of subjects.
Evaluated by flow cytometry.
Time frame: Day 3 - Day 45
Comparison of the number of the other cell types derived from the human adipose tissue-hematopoietic stem cells between both groups of grafted mice.
Evaluated by flow cytometry.
Time frame: Day 3 - Day 45
Comparison of the phenotype of the other cell types derived from the human adipose tissue-hematopoietic stem cells between both groups of grafted mice.
Evaluated by flow cytometry.
Time frame: Day 3 - Day 45
Comparison of the expression of genes coding for human inflammatory molecules in the adipose tissue of transplanted mice.
Evaluated by Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction. Main parameters analysed : Interleukin-6, Interleukin-1béta, Plasminogen activator inhibitor-1, Monocyte Chemoattractant Protein-1.
Time frame: Day 3 - Day 45
Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's glycemia.
According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects).
Time frame: Day 3 - Day 45
Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's insulinemia.
According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects).
Time frame: Day 3 - Day 45
Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's oral glucose tolerance test.
According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects).
Time frame: Day 3 - Day 45
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