Type2 Diabetes
Conditions
Keywords
Type 2 diabetes, Adipose tissue, Hematopoiesis, Inflammation, Macrophages
Brief summary
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.
Detailed description
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.
Interventions
PROCEDUREAbdominal subcutaneous biopsies and Blood test
Abdominal subcutaneous biopsies and blood test for each volunteer.
Sponsors
University Hospital, Toulouse
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
BASIC_SCIENCE
Masking
NONE
Masking description
All laboratory analyses will be performed blindly.
Intervention model description
20 male volunteers: 10 overweight/obese subjects with T2D and 10 healthy volunteers, age-matched. All volunteers will be treated in the same way.
Eligibility
Sex/Gender
MALE
Age
37 Years to 63 Years
Healthy volunteers
Yes
Inclusion criteria
* No major weight variation for at least 3 months * Biological assessment without clinically significant anomaly from the point of view of the investigator. * Acceptance of constraints related to participation in the study * Acceptance of participation in the constitution of a cell bank, a tissue bank and a serum library. * Affiliation to a social security scheme. Type 2 diabetic subjects: * Type 2 diabetes (discovered after the age of 35 years, without inaugural ketosis and absence of insulin therapy during the first year). * 40 to 60 year-old. * BMI between 27 and 35 kg / m² (included). * Treated by modification of lifestyle alone or associated to oral anti-diabetic therapy only. * With stable oral anti-diabetic treatment for at least 3 months. * HbA1c ≤ 8.5%. Healthy Volunteers: * BMI between 23 and 27 kg / m² (included). * 37 to 63 year-old, age-matched to a type 2 diabetes subject ± 5 years. * Fasting blood glucose \< 1,10 g / L. * HbA1c within normal limits (4 to 6%).
Exclusion criteria
* Excessive chronic alcohol consumption (\> 30 g / day or 210 g / week). * Tobacco consumption\> 10 cigarettes / day that cannot be stopped for 24 hours. * Anti-diabetic treatments that require sub-cutaneous injections * History of chronic or acute hematological pathology. * Systemic or acute inflammatory pathology. * Treatment with antiplatelet agents, non-steroidal anti-inflammatory drugs, glucocorticoids (excluding eye drops and sprays), or other immunosuppressive drugs. * History of cancer (except basal cell carcinoma). * Allergy to xylocaine or one of its derivatives. * Any significant pathology at the discretion of the investigator. * Any biological anomaly at the discretion of the investigator. * Positive human immunodeficiency virus serology. * Positive hepatitis B serology. * Positive hepatitis C serology. * Glomerular filtration rate less than 60 ml / min * Aspartate aminotransferase or alanine aminotransferase higher than 2.5-fold the upper normal value. * Hypertriglyceridemia \> 2.5 g / l * Person under the protection of justice, guardianship or curators. * Subject involved in another research protocol or in an exclusion period from another research protocol. * Cognitive disorder or mental pathology (at the discretion of the investigator).
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Number of pro-inflammatory macrophages derived from human adipose tissue in mice transplanted with adipose tissue-hematopoietic stem cells . | Day 3 - Day 45 | 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. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Comparison of the number of the other cell types derived from the human adipose tissue-hematopoietic stem cells between both groups of grafted mice. | Day 3 - Day 45 | Evaluated by flow cytometry. |
| Comparison of the phenotype of the other cell types derived from the human adipose tissue-hematopoietic stem cells between both groups of grafted mice. | Day 3 - Day 45 | Evaluated by flow cytometry. |
| Comparison of the expression of genes coding for human inflammatory molecules in the adipose tissue of transplanted mice. | Day 3 - Day 45 | 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. |
| Comparison of hematopoietic activity in vitro between both groups of subjects. | Day 3 - Day 45 | Evaluated by flow cytometry. |
| Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's insulinemia. | Day 3 - Day 45 | According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects). |
| Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's oral glucose tolerance test. | Day 3 - Day 45 | According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects). |
| Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's glycemia. | Day 3 - Day 45 | According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects). |
Countries
France
Outcome results
None listed