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Comparison of Hematopoietic Stem Cell Activity in Adipose Tissue From Type 2 Diabetic Patients and Healthy Volunteers (WAT2DO)

Primary Purpose

Type2 Diabetes

Status
Completed
Phase
Not Applicable
Locations
France
Study Type
Interventional
Intervention
Abdominal subcutaneous biopsies and Blood test
Sponsored by
University Hospital, Toulouse
About
Eligibility
Locations
Arms
Outcomes
Full info

About this trial

This is an interventional basic science trial for Type2 Diabetes focused on measuring Type 2 diabetes, Adipose tissue, Hematopoiesis, Inflammation, Macrophages

Eligibility Criteria

37 Years - 63 Years (Adult)MaleAccepts Healthy Volunteers

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).

Sites / Locations

  • CHU de Toulouse - Rangueil

Arms of the Study

Arm 1

Arm Type

Experimental

Arm Label

Patients

Arm Description

'Abdominal subcutaneous biopsies and Blood test'

Outcomes

Primary Outcome Measures

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.

Secondary Outcome Measures

Comparison of hematopoietic activity in vitro between both groups of subjects.
Evaluated by flow cytometry.
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.
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.
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.
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).
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).
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).

Full Information

First Posted
July 27, 2017
Last Updated
April 1, 2019
Sponsor
University Hospital, Toulouse
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1. Study Identification

Unique Protocol Identification Number
NCT03260452
Brief Title
Comparison of Hematopoietic Stem Cell Activity in Adipose Tissue From Type 2 Diabetic Patients and Healthy Volunteers
Acronym
WAT2DO
Official Title
Comparison of Hematopoietic Stem Cell Activity in Adipose Tissue From Type 2 Diabetic Patients and Healthy Volunteers: Proof of Concept Study. White Adipose Tissue and Type 2 Diabetes Onset: WAT2DO
Study Type
Interventional

2. Study Status

Record Verification Date
April 2019
Overall Recruitment Status
Completed
Study Start Date
June 28, 2018 (Actual)
Primary Completion Date
January 24, 2019 (Actual)
Study Completion Date
January 24, 2019 (Actual)

3. Sponsor/Collaborators

Responsible Party, by Official Title
Sponsor
Name of the Sponsor
University Hospital, Toulouse

4. Oversight

Studies a U.S. FDA-regulated Drug Product
No
Studies a U.S. FDA-regulated Device Product
No
Data Monitoring Committee
No

5. Study Description

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.

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Type2 Diabetes
Keywords
Type 2 diabetes, Adipose tissue, Hematopoiesis, Inflammation, Macrophages

7. Study Design

Primary Purpose
Basic Science
Study Phase
Not Applicable
Interventional Study Model
Single Group Assignment
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.
Masking
None (Open Label)
Masking Description
All laboratory analyses will be performed blindly.
Allocation
N/A
Enrollment
20 (Actual)

8. Arms, Groups, and Interventions

Arm Title
Patients
Arm Type
Experimental
Arm Description
'Abdominal subcutaneous biopsies and Blood test'
Intervention Type
Procedure
Intervention Name(s)
Abdominal subcutaneous biopsies and Blood test
Intervention Description
Abdominal subcutaneous biopsies and blood test for each volunteer.
Primary Outcome Measure Information:
Title
Number of pro-inflammatory macrophages derived from human adipose tissue in mice transplanted with adipose tissue-hematopoietic stem cells .
Description
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
Secondary Outcome Measure Information:
Title
Comparison of hematopoietic activity in vitro between both groups of subjects.
Description
Evaluated by flow cytometry.
Time Frame
Day 3 - Day 45
Title
Comparison of the number of the other cell types derived from the human adipose tissue-hematopoietic stem cells between both groups of grafted mice.
Description
Evaluated by flow cytometry.
Time Frame
Day 3 - Day 45
Title
Comparison of the phenotype of the other cell types derived from the human adipose tissue-hematopoietic stem cells between both groups of grafted mice.
Description
Evaluated by flow cytometry.
Time Frame
Day 3 - Day 45
Title
Comparison of the expression of genes coding for human inflammatory molecules in the adipose tissue of transplanted mice.
Description
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
Title
Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's glycemia.
Description
According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects).
Time Frame
Day 3 - Day 45
Title
Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's insulinemia.
Description
According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects).
Time Frame
Day 3 - Day 45
Title
Comparison of the metabolic profile of the transplanted mice evaluated by the grafted mice's oral glucose tolerance test.
Description
According to the origin of the grafted hematopoietic stem cells (diabetic or healthy voluntary subjects).
Time Frame
Day 3 - Day 45

10. Eligibility

Sex
Male
Minimum Age & Unit of Time
37 Years
Maximum Age & Unit of Time
63 Years
Accepts Healthy Volunteers
Accepts Healthy Volunteers
Eligibility Criteria
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).
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
Pierre GOURDY
Organizational Affiliation
CHU Toulouse
Official's Role
Principal Investigator
Facility Information:
Facility Name
CHU de Toulouse - Rangueil
City
Toulouse
ZIP/Postal Code
31059
Country
France

12. IPD Sharing Statement

Plan to Share IPD
No
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Comparison of Hematopoietic Stem Cell Activity in Adipose Tissue From Type 2 Diabetic Patients and Healthy Volunteers

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