Molecular Transducers of Physical Activity Consortium

ADULT PARTICIPANT INCLUSION CRITERIA - SEDENTARY PARTICIPANTS

   - Willingness to provide informed consent to participate in the MoTrPAC Study

   - Must be able to read and speak English well enough to provide informed consent and
   understand instructions

   - Aged ≥18 y

   - Body Mass Index (BMI) ≥19 to ≤35 kg/m2

   - Sedentary defined as self-reporting no more than 1 day per week, lasting no more than
   60 minutes, of regular (structured) EE [e.g., brisk walking, jogging, running,
   cycling, elliptical, or swimming activity that results in feelings of increased heart
   rate, rapid breathing, and/or sweating] or RE (resulting in muscular fatigue) in the
   past year

      - Persons bicycling as a mode of transportation to and from work >1 day/week etc.
      are not considered sedentary

      - Leisure walkers are included unless they meet the heart rate, breathing, and
      sweating criteria noted above

      - Willingness to include de-identified individual-level data at low risk of
      re-identification (e.g.,non-genomic data) in the MoTrPAC open-access database

      - Only one member of a household can participate

ADULT PARTICIPANT INCLUSION CRITERIA - HIGHLY ACTIVE PARTICIPANTS

   - Willingness to provide informed consent to participate in the MoTrPAC Study

   - Must be able to read and speak English well enough to provide informed consent and
   understand instructions

   - Aged ≥18 y

   - BMI ≥19 to ≤35 kg/m2

   - Comparator Participants

      - HAEE: defined as ≥240 minutes/week of ET for ≥1 year; this can include running,
      walking (brisk, power), cycling, elliptical, etc. which (at a minimum) results in
      increased heart rate, rapid breathing and sweating

      - Must include cycling at least 2 days/week

      - RT in the past year must be limited to ≤2 days/week of upper body RE and ≤2
      muscle groups of upper body RE and ≤1 day/week of lower body RE

      - HARE: defined as RT of ≥3 upper and ≥3 lower body muscle groups ≥2 times/week for
      ≥1 year; using a prescription sufficient to increase strength and muscle mass

      - ET in the past year must be limited to ≤90 minutes/week of vigorous EE, with no
      limit on cycling days per week

      - Elite or Competitive Athletes: can be included, if they meet HAEE or HARE
      inclusion criteria

      - Potential participants are informed that use of performance enhancing drugs in
      the last 6 months is exclusionary

      - Willingness to include de-identified individual-level data at low risk of
      re-identification (e.g., non-genomic data) in the MoTrPAC open-access database

      - In addition to meeting HAEE or HARE inclusion criteria, all HA participants must
      meet all other exclusion criteria defined in this protocol

EXCLUSION CRITERIA

ADULT PARTICIPANT EXCLUSION CRITERIA Exclusion criteria are confirmed by self-report (i.e.,
medical and medication histories reviewed by a clinician), screening tests performed by the
MoTrPAC study team at each clinical site, and/or clinician judgement as specified for each
criterion.

   - Diabetes (self-report and screening tests)

      - Treatment with any hypoglycemic agents (self-report) or A1c >6.4% (screening
      test; may reassess once if 6.5-6.7%)

      - Fasting glucose >125 mg/dL (screening test; may reassess once)

      - Use of hypoglycemic drugs for non-diabetic reasons (self-report)

   - Abnormal bleeding or coagulopathy (self-report)

      - History of a bleeding disorder or clotting abnormality

   - Thyroid disease (screening test)

      - Thyroid Stimulating Hormone (TSH) value >5.9 IU/mL

      - Individuals with hypothyroidism may be referred to their primary care provider
      (PCP) for evaluation and retested; any medication change must be stable for ≥3
      months prior to retesting

      - Individuals with hyperthyroidism are excluded, including those with normal TSH on
      pharmacologic treatment

   - Pulmonary (self-report)

      - Clinical diagnosis of Chronic Obstructive Pulmonary Disease (COPD)

   - Metabolic bone disease (self-report)

      - History of non-traumatic fracture from a standing height or less

      - Current pharmacologic treatment for low bone mass or osteoporosis, other than
      calcium, vitamin D, or estrogen

   - Estrogens, progestins (self-report)

      - Supplemental, replacement or therapeutic use of estrogens or progestins within
      the last 6 months, other than birth control or to control menopausal symptoms

   - Pregnancy (screening test) and pregnancy-related conditions (self-report)

      - Pregnant - pregnancy test performed on day of DXA scan in women of child-bearing
      potential

      - Post-partum during the last 12 months

      - Lactating during the last 12 months

      - Planning to become pregnant during the participation period

   - Elevated blood pressure readings (screening test)

      - Resting Systolic Blood Pressure (SBP) ≥150 mmHg or Resting Diastolic Blood
      Pressure (DBP) ≥95 mmHg

      - Reassessment of BP during screening will be allowed to ensure rested values are
      repeatable

   - Cardiovascular (self-report, screening test, and clinician judgement)

      - Congestive heart failure, coronary artery disease, significant valvular disease,
      congenital heart disease, serious arrhythmia, stroke, or symptomatic peripheral
      artery disease (self-report, screening test)

      - Specific criteria used to determine whether a volunteer can undergo the screening
      CPET follow the American Heart Association (AHA) Criteria [54]

      - Inability to complete the CPET

      - Reassessment of the CPET may be allowed under some circumstances (e.g., test was
      not a maximal effort)

   - Abnormal blood lipid profile (screening test)

      - Fasting triglycerides >500 mg/dL

      - Low-density lipoprotein cholesterol (LDL-C) >190mg/dL

   - Cancer (self-report)

      - History of cancer treatment (other than non-melanoma skin cancer) and not
      "cancer-free" for at least 2 years

      - Anti-hormonal therapy (e.g., for breast or prostate cancer) within the last 6
      months

   - Chronic active or latent infection (self-report)

      - Active or latent infections requiring chronic antibiotic or anti-viral treatment

      - Chronic active infection whether on chronic antimicrobials or not

      - Human Immunodeficiency Virus

      - Active hepatitis B or C undergoing antiviral therapy

      - Individuals successfully treated for hepatitis C and virologically negative for
      at least 6 months are not excluded

   - Liver enzyme tests (Alanine transaminase, Aspartate transaminase) (screening test)

      - >2 times the laboratory upper limit of normal

      - Reassessment during screening may be allowed under some conditions (e.g., recent
      use of acetaminophen)

      - Individuals may be referred to their PCP for evaluation; any medication change
      must be stable for ≥3 months prior to retesting

   - Chronic renal insufficiency (screening test)

      - Estimated glomerular filtration rate <60 mL/min/1.73 m2 from serum creatinine
      (mg/dL) by the Chronic Kidney Disease Epidemiology Collaboration equation

      - Reassessment may be allowed under some conditions (e.g., questionable hydration
      status or other acute renal insult)

   - Hematocrit (screening test)

      - Hematocrit >3 points outside of the local normal laboratory ranges for women and
      men

      - Reassessment may be allowed under certain conditions

      - Individuals may be referred to their PCP for evaluation; any medication change
      must be stable for ≥3 months prior to retesting

      - Individuals with known thalassemia trait may be included (despite having >3
      points outside of the local normal laboratory ranges), upon approval from their
      PCP or a hematologist

   - Blood donation (self-report)

      - Whole blood donation in the last 3 months or plans for blood donation during the
      entire protocol period

      - Platelet or plasma donation in the last week or plans for platelet or plasma
      donation during the entire protocol period

   - Autoimmune disorders (self-report)

      - Individuals receiving any active treatment (including monoclonal antibodies)
      within the last 6 months

   - Alcohol consumption (self-report)

      - More than 7 drinks per week for women

      - More than 14 drinks per week for men

      - History of binge drinking (≥5 drinks for males or ≥4 drinks for females in a
      2-hour period more than once per month)

   - Tobacco (self-report)

      - Current smokers: any tobacco or e-cigarette/e-nicotine products

      - Former smokers:

      - Stopped smoking <10 years at time of screening for those with a ≥20 pack-year
      smoking history

      - Stopped smoking <5 years at time of screening for those with a <20 pack-year
      smoking history

   - Marijuana (self-report)

      - Self-reported use ≥3 days/week in any form

   - Shift workers (self-report)

      - Night shift work in the last 6 months

      - Planning night shift work during the study period

   - Cognitive status (screening)

      - Unable to give consent to participate in and safely complete the protocol, as
      based on the judgement of the local investigator

   - Psychiatric illness (self-report and screening test)

      - Hospitalization for any psychiatric condition within one year (self-report)

      - Center for Epidemiological Studies-Depression Scale (CESD) score ≥16 [55]
      (screening test)

   - Weight change (self-report)

      - Weight change (intentional or not) over the last 6 months of >5% of body weight

      - Plan to lose or gain weight during the study

   - Lidocaine or other local anesthetic (self-report)

      - Known allergy to lidocaine or other local anesthetic

   - COVID-19 infection

      - Hospitalization for COVID-19 infection in the past 12 months

      - Individuals who tested positive for COVID-19 but were not hospitalized must be
      symptom-free at least 14 days

   - Other (clinician judgement)

      - Genetic metabolic disorders that could effect metabolomic results
      (e.g.,phenylketonuria)

      - Any other cardiovascular, pulmonary, orthopedic, neurologic, psychiatric.
      metabolic, or other conditions that, in the opinion of the local clinician, would
      preclude participation and successful completion of the protocol

      - Any other illnesses that, in the opinion of the local clinician, would negatively
      impact or mitigate participation in and completion of the protocol

EXCLUSIONS FOR MEDICATION USE

   - Continuous use for 7 days or more of a new drug (prescription or over-the-counter;
   additional guidance in the MOP) in the last 3 months; eye and ear drops are allowed
   regardless of when they were started

   - Dose change for any chronic-use drug in the last 3 months; changes in eye and ear
   drops are allowed

   - Cardiovascular

      - Beta blockers and centrally acting anti-hypertensive drugs

      - Anticoagulants

      - Antiarrhythmic drugs

      - Antiplatelet drugs (other than aspirin ≤100 mg/day)

   - Psychiatric drugs

      - Chronic use of medium or long-acting sedatives and hypnotics including all
      benzodiazepines; (short-acting non-benzodiazepine sedative-hypnotics are allowed)

      - Mood stabilizers

      - Antiepileptic drugs

      - Stimulants, Attention-Deficit/Hyperactivity Disorder (ADHD) drugs

      - Anti-psychotic drugs

   - Pulmonary, inflammation

      - Chronic oral steroids; inhaled steroids are allowed

      - Burst/taper oral steroids more than once in the last 12 months; inhaled steroids
      are allowed

      - B2-agonists

      - allowed if on stable dose at least 3 months

   - Genitourinary

      - 5-alpha reductase inhibitors

      - Daily phosphodiesterase type 5 inhibitor use

   - Hormonal

      - Androgenic anabolic steroids

      - Anti-estrogens, anti-androgens

      - Estrogens and/or progestins used for reasons other than birth control or
      menopause

      - Growth hormone, insulin like growth factor-I, growth hormone releasing hormone

      - Any drugs used to treat diabetes mellitus or to lower blood glucose

      - Metformin for any indication

      - Any drugs used specifically to induce weight loss

      - Any drugs used specifically to induce muscle growth/hypertrophy or augment
      exercise-induced muscle hypertrophy

   - Pain/inflammation

      - Narcotics and narcotic receptor agonists

      - Regular use of non-steroidal anti-inflammatory drugs (NSAIDs) or acetaminophen

      - Muscle relaxants ≥3 days per week

      - Oral/sublingual cannabidiol or similar in any formulation

   - Other

      - Chronic systemic antimicrobials (antibiotic, antiviral, antifungal, antiparasite,
      etc) for any reason

      - High-potency topical steroids if ≥10% of surface area using rule of 9s

      - Continuous/chronic use of antibiotics or other anti-infectives for treatment or
      prevention

      - Monoclonal antibodies

      - Anti-rejection medications/immune suppressants

   - Any other medications that, in the opinion of local clinicians, would negatively
   impact or mitigate full participation and completion