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

Interventional Clinical Trial

Official title:

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

Clinical Trial Details — Status: Active, not recruiting

Administrative data

• NCT number: NCT05554653
• Other study ID #: ULE
• Status: Active, not recruiting
• Phase: N/A
• Start date: October 1, 2022
• Est. completion date: August 2024

Study information

• Verified date: November 2023
• Source: University of Toronto
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

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.

Recruitment information / eligibility

• Status: Active, not recruiting
• Enrollment: 8
• Est. completion date: August 2024
• Est. primary completion date: October 3, 2023
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: All
• Age group: 18 Years to 35 Years

Eligibility

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).

Related Conditions & MeSH terms

• Interventional

Intervention

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.
• Carbohydrate Placebo
iso-caloric placebo (4g maltodextrin). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda.

Locations

Country	Name	City	State
Canada	Goldring Centre for High Performance Sport at the University of Toronto	Toronto	Ontario

Sponsors (1)

Lead Sponsor	Collaborator
Daniel Moore

Country where clinical trial is conducted

Canada, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Other	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
Primary	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	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
Secondary	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
Secondary	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