Influence of Leucine Enriched Amino Acids on Myofibrillar and Collagen Protein Synthesis

Influence of Leucine Enriched Amino Acids on Myofibrillar and Collagen Protein Synthesis in Human Skeletal Muscle

The primary objective will be to determine the effect of leucine-enriched essential amino acids (LEAA) compared to carbohydrate placebo on dietary incorporation of [D5] Phenylalanine & [D5] Glycine into the three skeletal muscle protein pools (myofibrillar, sarcoplasmic and collagen), both following resistance exercise and at rest, with the two tracers provided as a 'intrinsically labeled' bolus. Other outcomes will relate to molecular regulation of protein synthesis.

Study Overview

• Status: Recruiting
• Conditions: Interventional
• Intervention / Treatment: Dietary supplement: Leucine enriched amino acids; Dietary supplement: Carbohydrate Placebo

• Study Type: Interventional
• Enrollment (Anticipated): 10
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Daniel Moore, PhD; Phone Number: 416-946-; Email: dr.moore@utoronto.ca

Study Locations

• Canada
  • Ontario
    • Toronto, Ontario, Canada, M5S2C9
      • Recruiting
      • Goldring Centre for High Performance Sport at the University of Toronto

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 35 years (ADULT)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Healthy, male and female, recreationally-active participants.
• Healthy will be defined as screened by the PAR-Q+ (The Physical Activity Readiness Questionnaire for everyone; Appendix B).
• Self-reported not having engaged in resistance exercise and/or lower body plyometrics for at least 3 months prior to the study.
• Participants will be aged 18-35 years old.
• Participants are willing to abide by the compliance rules of this study.
• Self-reported regular menstrual cycle (25-35d) within the last 3 months (female participants).

Exclusion Criteria:

• Inability to adhere to any of the compliance rules judged by principle investigator or medical doctor.
• Self-reported regular tobacco use.
• Self-reported illicit drug use (e.g. growth hormone, testosterone, etc.).
• Individuals who have participated in studies within the past year involving any of the stable isotopes in the study.
• Use of birth control and discontinued use within the last 3 months (female participants).

Study Plan

How is the study designed?

Design Details

• Primary Purpose: TREATMENT
• Allocation: RANDOMIZED
• Interventional Model: CROSSOVER
• Masking: TRIPLE

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
ACTIVE_COMPARATOR: Leucine enriched essential amino acids; Leucine enriched essential amino acids (4g total, of which 1.6g are leucine). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda to mask taste.	Dietary supplement: Leucine enriched amino acids; Leucine enriched essential amino acids (4g total, of which 1.6g are leucine). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda.; Other Names: LEAA
PLACEBO_COMPARATOR: Carbohdyrate Placebo; Iso-caloric placebo (4g maltodextrin to replace LEAA). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda to mask taste.	Dietary supplement: Carbohydrate Placebo; iso-caloric placebo (4g maltodextrin). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Myofibrillar Protein synthesis	Rate of incorporation of orally ingested amino acid tracers (D5 Phe and D5 Gly) into myofibrillar proteins isolated from skeletal muscle biopsies 4h following drink (LEAA or PLA) consumption and resistance exercise or at rest. Enrichment will be quantified via tandem LC-MS/MS to derive a rate of incorporation over the 4h period.	4 hours

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Sarcoplasmic protein synthesis	Rate of incorporation of orally ingested amino acid tracers (D5 Phe and D5 Gly) into sarcoplasmic (i.e., soluble) proteins isolated from skeletal muscle biopsies 4h following drink consumption (LEAA or PLA) and resistance exercise or at rest. Enrichment will be quantified via tandem LC-MS/MS.	4 hours
Collagen protein synthesis	Rate of incorporation of orally ingested amino acid tracers (D5 Phe and D5 Gly) into collagen (i.e., non-soluble) proteins isolated from skeletal muscle biopsies 4h following drink consumption (LEAA or PLA) and resistance exercise or at rest. Enrichment will be quantified via tandem LC-MS/MS.	4 hours
Protein localization via immunofluorescence	Antibody-determined Phosphorylated RPS6 (S240/244) and mTOR protein localization in human muscle cross sections and longitudinal sections as examined via immunofluorescence. Muscle sections (both longitudinal and cross-sections) will be derived from snap-frozen muscle biopsies performed 4h following drink consumption (LEAA or PLA) with or without resistance exercise (unilateral). Localization will be performed as done by Hodson et al., (2022; AJP Cell). Particularly, the localization of these targets relative to the cell periphery and z-discs (longitudinal sections only) will be analyzed via colocalization analysis.	4 hours

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Isolation of primary myoblasts for in vitro nutritional experiments	isolation of primary myoblasts from skeletal muscle biopsies for growth in vitro and nutritional experiments in cell culture models	6-9 months

Collaborators and Investigators

• Sponsor: Daniel Moore

Study record dates

Study Major Dates

• Study Start (ACTUAL): October 1, 2022
• Primary Completion (ANTICIPATED): September 1, 2023
• Study Completion (ANTICIPATED): August 1, 2024

Study Registration Dates

• First Submitted: September 19, 2022
• First Submitted That Met QC Criteria: September 22, 2022
• First Posted (ACTUAL): September 26, 2022

Study Record Updates

• Last Update Posted (ACTUAL): February 2, 2023
• Last Update Submitted That Met QC Criteria: January 31, 2023
• Last Verified: January 1, 2023

More Information

Terms related to this study

• Keywords: Leucine-enriched amino acids; Carbohydrate Placebo
• Other Study ID Numbers: ULE (Other Identifier: Universidad de León)

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No