Single Nuclei RNA-seq to Map Adipose Cellular Populations and Senescent Cells in Older Subjects

Study Overview
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Brief Summary
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All participants will undergo baseline biopsies of subcutaneous abdominal adipose tissue for cellular/molecular profiling via snRNA-seq and metabolic/physiological assessments (insulin sensitivity, glucose tolerance, and β-cell function). Older obese participants will be randomized into three arms: lifestyle intervention (n=24), senolytics (n=24), or placebo (n=24).

Detailed Description
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All participants after consent and enrollment will undergo adipose tissue single nuclei RNA sequencing and metabolic phenotyping. Subjects will undergo glucose tolerance testing to document glucose tolerance status. Each subject will be provided an accelerometer to be worn on their dominant wrist for 7 days for assessment of habitual activity. A dietitian will teach them to utilize the SmartIntake3 smartphone food picture application (app) for a 7-day food record. The app will be used to record amount of each meal consumed in order to determine daily food and beverage and supplement intake and quantity for dietary composition analysis. DEXA analysis will be performed to measure lean and fat body mass. Subjects will undergo evaluation of physical function/performance, including the Short Physical Performance Battery (SPPB) and VO2 peak testing for assessment of aerobic capacity. The SPPB will be done in older adults only. The NIH Patient-Reported Outcomes Measurement System (PROMIS) will be used to measure participants' self-report of symptoms, function, and health-related quality of life in the domains of physical, mental and social health. All subjects will undergo a two-step euglycemic insulin clamp and indirect calorimetry. Only older obese participants will continue to the randomization to likestyle intervention, senolytic agents or placebo.

Official Title
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Single Nuclei RNA-sequencing to Map Adipose Cellular Populations and Senescent Cells in Older Subjects

Conditions
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Obesity, Healthy Lifestyle

Intervention / Treatment
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* Other: Lifestyle Intervention
* Drug: Dasatinib 100 MG
* Drug: Quercetin 1000mg
* Drug: Placebo
* Procedure: Abdominal adipose tissue biopsy


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Both Sexes Age: younger lean group 18-30 years with BMI 18.5-24.9 kg/m2; older lean group ≥ 65 years with BMI All races and ethnic groups Community dwelling Sedentary (≤1.5 h of exercise per week) Nondiabetic (fasting plasma glucose < 126 mg/dl, 2-h glucose during oral glucose tolerance test (OGTT) < 140mg/dl, and A1c < 6.5% For all female participants who are women of childbearing potential (WOCBP), who are not pregnant or breast feeding, at least one of the following conditions must apply: A documented hysterectomy, bilateral salpingectomy, or bilateral oophorectomy Use of a contraceptive method that is highly effective (with a failure rate of <1% per year), preferably with low user dependency (implantable progesterone-only hormone contraception, intrauterine hormone releasing system, bilateral tubal occlusion, vasectomized partner) during the intervention period of the study and for at least 30 days after the last dose of study intervention to eliminate any reproductive safety risk of the study drug. Use of a contraceptive method that is highly effective (with a failure rate of <1% per year), with high user dependency, (oral/intravaginal/injectable combined estrogen and progesterone contraception, oral/injectable progesterone only hormone contraception, sexual abstinence) during the intervention period and for at least 30 days after the last dose of study intervention to eliminate any reproductive safety risk of the study drug. In addition to the highly effective methods: male or female condom with or without spermicide; cervical cap, diaphragm, or sponge with spermicide; a combination of male condom with either cervical cap, diaphragm, or sponge with spermicide. ECG value after 10 minutes of resting in the supine position in the following ranges: 120ms<PR<220ms: QRS<120ms; QTc<430ms for males and QTc<450ms for females and normal ECG tracing, unless the investigator considers the ECG abnormality to be not clinically relevant. Exclusion Criteria: Diabetes, clinically diagnosed or HbA1c > 6.5% and/or fasting plasma glucose > 126 mg/dl and/or use of anti-diabetic medications. Participating in > 1.5 h of structured exercise/week Unstable weight (>3% change in last 3 months) Neurological, musculoskeletal, or other conditions that may limit subject's ability to complete study physical assessment and training Active autoimmune/inflammatory disease including: rheumatoid arthritis, multiple sclerosis, systemic lupus erythematous, inflammatory bowel disease Laboratory parameters outside the normal range: impaired kidney function (eGFR < 30ml/min/1.73m² as calculated by the CKD-EPI equation); impaired liver function (AST or ALT level > 2 times upper limit of normal (ULN); total Bilirubin level > 1.5 times ULN; TSH > 1.5 times ULN or < lower limit of normal (LLN); Hemoglobin <10.0 g/dl; Platelets <125,000 cell/mm³; Platelets < 125,000 cell/mm³ Prothrombin time (PT) > 1.0 times ULN Partial prothrombin time (PTT) > 1.0 times ULN. Active gastrointestinal disease; coagulopathy; GI bleed within 6 months Clinically significant heart disease (e.g. NYH Classification >II; ischemia) Peripheral vascular disease (claudication) QTc prolongation >45 msec Use of anti-arrhythmic medications known to cause QTc prolongation, anti-platelet or anti-coagulant medication (see section 5.3) Use of quinolone antibiotics or any other drugs that may prolong the QTc interval (see section 5.3) Pulmonary disease (COPD), severe asthma or exercise-induced asthma Recent systemic or pulmonary embolus Uncontrolled blood pressure (systolic BP>170, diastolic BP>95 mmHg) Smoking, alcohol use (history of regular alcohol consumption exceeding 7 drinks/week for female participants or 14 drinks/week for male participants. 1 drink = 5 ounces [150ml] of wine or 12 ounces [360ml] of beer or 1.5 ounces [45ml] of hard liquor) or recreational drug use Pregnant or breastfeeding Postmenopausal women new (within 6 months) to systemic hormone replacement therapy Previous bariatric surgery History of stroke with motor disability Recent (3 years) treated cancer other than basal cell carcinoma Acute or chronic infection Medication that might interfere with metabolic studies (weight loss medication, systemic steroids, immunosuppressants) within 6 months (see section 5.3) Potentially senolytic agents within the last 6 months: fisetin, quercetin, luteolin, dasatinib, piperlongumine, or navitoclax (see section 5.3) History of allergy to dasatinib, quercetin and/or lidocaine.

Ages Eligible for Study
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Minimum Age: 18 Years

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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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: All participants will undergo cellular/molecular profiling and older obese participants will be randomized into 3 arms.
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Other: Younger Lean Group<br>Participants will be aged 18-30 years and have a BMI of 18.5 - 24.9 kg/m2 | Procedure: Abdominal adipose tissue biopsy<br>* All participants will undergo baseline biopsies of subcutaneous abdominal adipose tissue for cellular/molecular profiling via snRNA-seq<br>* Other names: Subcutaneous abdominal adipose tissue biopsy;|
| Other: Older Lean Group<br>Participants will be over 65 years of age with a BMI of 18.5 to 24.9 kg/m2 | Procedure: Abdominal adipose tissue biopsy<br>* All participants will undergo baseline biopsies of subcutaneous abdominal adipose tissue for cellular/molecular profiling via snRNA-seq<br>* Other names: Subcutaneous abdominal adipose tissue biopsy;|
| Experimental: Older Obese Group<br>Participants will be over 65 years of age with a BMI of 30-39.9 kg/m2. | Other: Lifestyle Intervention<br>* Exercise: Subjects will undergo a combined aerobic and resistance exercise intervention. The investigators propose a multi-modal training program because they improve metabolic outcomes and adding resistance training reduces the risk of muscle loss during weight loss. Subjects will exercise in the gym for three sessions per week for 10 weeks under supervision of an exercise physiologist. On each session, aerobic exercise will be followed by resistance exercise. Diet: The aim of the dietary intervention is to reduce caloric intake sufficient to result in 8-10% weight loss, while incorporating behavioral and dietary strategies to maximize adherence and to minimize risk of muscle and bone loss. Subjects will attend small-group sessions led by a dietitian for changing their dietary composition to follow American Heart Association (AHA)/American College of Cardiology (ACC) guidelines.<br>* Other names: Exercise and Diet;Drug: Dasatinib 100 MG<br>* 100 mg of dasatinib (D) daily for 3 consecutive days plus quercetin (Q) for the same 3 consecutive days, followed by a 25-day (+/- 2 day) no-drug period to complete a single round. This will be repeated twice more until completing 3 rounds total in approximately 10 consecutive weeks.<br>* Other names: Sprycell;Drug: Quercetin 1000mg<br>* Quercetin (Q) (4) 250 mg capsules daily (total 1000 mg daily) plus dasatinib (D) same 3 consecutive days, followed by a 25-day (+/- 2 day) no-drug period to complete a single round. This will be repeated twice more until completing 3 rounds total in approximately 10 consecutive weeks.<br>Drug: Placebo<br>* Subjects will receive a placebo (methylcellulose) to provide a similar mass, number of tablets/ capsules, and frequency as the senolytic arm.<br>* Other names: Placebo capsule;Procedure: Abdominal adipose tissue biopsy<br>* All participants will undergo baseline biopsies of subcutaneous abdominal adipose tissue for cellular/molecular profiling via snRNA-seq<br>* Other names: Subcutaneous abdominal adipose tissue biopsy;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Insulin sensitivity | Change in skeletal muscle insulin sensitivity | Week 4 and Week 15 | 
| Glucose tolerance | Measurement of change in glucose tolerance using a glucose tolerance test | Baseline to Week 14 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Senescence-associated secretory phenotype (SASP) | Measurement of change of SASP in adipose tissue | Baseline to Week 16 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Lifestyle intervention, Single RNA sequencing, Cell Senescence