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Completed

NCT Number: NCT05457218

Effect of Insect-derived Protein Supplements on Protein Synthesis and mTORC1 Activation

The purpose of this study is to determine the ability of insect-derived protein supplements (IPC80 and Whole Buffalo Powder Ynsect®) to activate the mTORC1 pathway and protein synthesis in muscle cells.

To achieve this aim, participants will consume a supplemental dose (0.25 g/kg body weight) of IPC80,Whole Buffalo Powder or whey protein. Blood samples will be drawn before and 15, 30, 60 and 90 minutes after the supplementation.

Protein synthesis will be assessed using stably transfected C2C12 muscle cells with a plasmid (pcDNA3 luciferase) and to determine the ability of a protein source to activate mTORC1, stably transfected C2C12 muscle cells with a plasmid (pcDNA3-TOP luciferase) where the luciferase mRNA contains a 5'TOP. 5'TOP which is specifically regulated by mTORC1 activity will be used.

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Key information

Conditions

Effect of Food Supplement

Age range

18 year–30 year

Sex eligibility

All sexes

Study type

Interventional

Phase

Not applicable

Primary location

Hickey Laboratory

Davis, California, 95616, United States

About this study

The purpose of this study is to determine the ability of insect-derived protein supplements (IPC80 and Whole Buffalo Powder, Ynsect®) to activate the mTORC1 pathway and protein synthesis in muscle cells.

To achieve this aim, participants will consume a supplemental dose (0.25 g/kg body weight) of IPC80, Whole Buffalo Powder or whey protein. Blood samples will be drawn before and 15, 30, 60 and 90 minutes after the supplementation.

It is expected that this project will determine whether insect-derived protein supplements (IPC80 and Whole Buffalo Powder, Ynsect®) have the ability to induce activation of the mTORC1 pathway and protein synthesis in muscle cells. Results from this study will give further insight into the usage of this novel sustainable protein source to support muscle protein synthesis after exercise.

Male and female participants between the ages of 18-30 years of age will be enrolled in the study. The study is a randomized double-blind crossover design with neither the subjects nor the investigators knowing who is on which treatment (IPC80, Whole Buffalo Powder or Whey).

I. Baseline blood draw The subjects will arrive in the laboratory following an overnight fast. The antecubital vein will be cannulated, and an initial 5 mL baseline blood sample will be collected.

II. Supplementation After baseline blood draw, subjects will be provided with a supplement, 0.25 g/kg body weight of either whey protein, IPC80 or Whole Buffalo Powder dissolved in 250ml of water.

III. Postprandial blood draws Following consumption of the supplement, the participants will remain in the laboratory for another 1.5 hours, they may bring books or electronic devices to pass the time. Four more blood samples of 5 mL each will be obtained at 15, 30, 60, and 90 minutes after supplementation, totaling five blood draws of 5mL per visit (25mL per visit), and 75 mL of blood drawn in total for each subject completing the entire study.

Blood will be collected in 5 mL serum separating tubes. All blood samples will be allowed to clot for 1hr before centrifugation at 1000 x g for 10 minutes and the serum will be frozen and kept at -30°C until processed. Treatments will be randomized to avoid an order effect and a washout period of at least 48h between trials will be used to minimize the effect of the previous treatment.

To determine protein synthesis, we have stably transfected C2C12 muscle cells with a plasmid (pcDNA3 luciferase) that we use to measure global protein synthesis. C2C12Luc cells will be plated in 24-well plates and differentiated over 4 days. Differentiated C2C12 cells will be fasted by washing with PBS and then treating them with Test Media (20% DMEM; 80% Hank's buffered salt solution (HBSS)) for 15 minutes. Fasted muscle cells will then be treated with Test Media containing 0, 2.5, 5, or 10% baseline or fed serum for 60 minutes. Cells will be collected in passive lysis buffer and firefly luciferase activity will be determined as described previously (Philp et al., 2013).

To determine the ability of a protein source to activate mTORC1 we have stably transfected C2C12 muscle cells with a plasmid (pcDNA3-TOP luciferase) where the luciferase mRNA contains a 5' terminal polypyrimidine tract (TOP). 5'TOP mRNA are specifically regulated by mTORC1 activity (Jefferies et al., 1994). As above, differentiated C2C12TOPLuc muscle cells in 24-well plates will be stimulated using the baseline or fed serum as described above. The degree of mTORC1 activation will be determined as the difference in slopes between the baseline and fed sera. The induction of mTORC1 and protein synthesis by whey and IPC80 and Whole Buffalo Powder will be compared directly within individuals and then across groups.

Who can participate

Healthy volunteers accepted: Yes

Only the study team can determine whether someone qualifies for participation.

Inclusion criteria

  • Healthy active male or female
  • Normal weight (BMI between 18 and 25 kg/m2)

Exclusion criteria

  • Receiving any medication that may interfere with the study.
  • Health or dietary restrictions that would be affected by the supplementation protocol.
  • Allergy to shellfish (the chitins in the mealworm exoskeleton can activate shellfish allergies)

Treatment and study plan

Whey Protein

Dietary Supplement

The participant will consume a supplemental dose of whey protein (0.25 g/kg body weight)

IPC80

Dietary Supplement

The participant will consume a supplemental dose of IPC80 (0.25 g/kg body weight)

Whole Buffalo Powder

Dietary Supplement

The participant will consume a supplemental dose of Whole Buffalo Powder (0.25 g/kg body weight)

Primary outcomes

  1. Muscle protein synthesis

    Time frame: Baseline (0 hour) to 15 minutes after supplementation

    To measure muscle protein synthesis, stably transfected C2C12 muscle cells with a plasmid (pcDNA3 luciferase) will be used. C2C12Luc cells will be plated in 24-well plates and differentiated over 4 days. Differentiated C2C12 cells will be fasted by washing with PBS and then treating them with Test Media (20% DMEM) for 15 minutes. Fasted muscle cells will then be treated with Test Media containing 0, 2.5, 5, or 10% baseline or fed serum (from blood samples at 15, 30, 60, and 90 minutes after supplementation) for 60 minutes. Cells will be collected in passive lysis buffer and firefly luciferase activity will be determined.

  2. Muscle protein synthesis

    Time frame: Baseline (0 hour) to 30 minutes after supplementation

    To measure muscle protein synthesis, stably transfected C2C12 muscle cells with a plasmid (pcDNA3 luciferase) will be used. C2C12Luc cells will be plated in 24-well plates and differentiated over 4 days. Differentiated C2C12 cells will be fasted by washing with PBS and then treating them with Test Media (20% DMEM) for 15 minutes. Fasted muscle cells will then be treated with Test Media containing 0, 2.5, 5, or 10% baseline or fed serum (from blood samples at 15, 30, 60, and 90 minutes after supplementation) for 60 minutes. Cells will be collected in passive lysis buffer and firefly luciferase activity will be determined.

  3. Muscle protein synthesis

    Time frame: Baseline (0 hour) to 60 minutes after supplementation

    To measure muscle protein synthesis, stably transfected C2C12 muscle cells with a plasmid (pcDNA3 luciferase) will be used. C2C12Luc cells will be plated in 24-well plates and differentiated over 4 days. Differentiated C2C12 cells will be fasted by washing with PBS and then treating them with Test Media (20% DMEM) for 15 minutes. Fasted muscle cells will then be treated with Test Media containing 0, 2.5, 5, or 10% baseline or fed serum (from blood samples at 15, 30, 60, and 90 minutes after supplementation) for 60 minutes. Cells will be collected in passive lysis buffer and firefly luciferase activity will be determined.

  4. Muscle protein synthesis

    Time frame: Baseline (0 hour) to 90 minutes after supplementation

    To measure muscle protein synthesis, stably transfected C2C12 muscle cells with a plasmid (pcDNA3 luciferase) will be used. C2C12Luc cells will be plated in 24-well plates and differentiated over 4 days. Differentiated C2C12 cells will be fasted by washing with PBS and then treating them with Test Media (20% DMEM) for 15 minutes. Fasted muscle cells will then be treated with Test Media containing 0, 2.5, 5, or 10% baseline or fed serum (from blood samples at 15, 30, 60, and 90 minutes after supplementation) for 60 minutes. Cells will be collected in passive lysis buffer and firefly luciferase activity will be determined.

Secondary outcomes

  1. mTORC1 activity

    Time frame: Baseline (0 hour) to 15 minutes after supplementation

    To measure the ability of a protein source (whey protein, IPC80 or Whole Buffalo Powder) to activate mTORC1, stably transfected C2C12 muscle cells with a plasmid (pcDNA3-TOP luciferase) where the luciferase mRNA contains a 5'TOP. 5'TOP mRNA , which specifically regulated by mTORC1 activity, will be used. Differentiated C2C12TOPLuc muscle cells in 24-well plates will be stimulated using the baseline or fed serum (rom blood samples at 15, 30, 60, and 90 minutes after supplementation). The degree of mTORC1 activation will be determined as the difference in slopes between the baseline and fed sera.

  2. mTORC1 activity

    Time frame: Baseline (0 hour) to 30 minutes after supplementation

    To measure the ability of a protein source (whey protein, IPC80 or Whole Buffalo Powderl) to activate mTORC1, stably transfected C2C12 muscle cells with a plasmid (pcDNA3-TOP luciferase) where the luciferase mRNA contains a 5'TOP. 5'TOP mRNA , which specifically regulated by mTORC1 activity, will be used. Differentiated C2C12TOPLuc muscle cells in 24-well plates will be stimulated using the baseline or fed serum (rom blood samples at 15, 30, 60, and 90 minutes after supplementation). The degree of mTORC1 activation will be determined as the difference in slopes between the baseline and fed sera.

  3. mTORC1 activity

    Time frame: Baseline (0 hour) to 60 minutes after supplementation

    To measure the ability of a protein source (whey protein, IPC80 or Whole Buffalo Powder) to activate mTORC1, stably transfected C2C12 muscle cells with a plasmid (pcDNA3-TOP luciferase) where the luciferase mRNA contains a 5'TOP. 5'TOP mRNA , which specifically regulated by mTORC1 activity, will be used. Differentiated C2C12TOPLuc muscle cells in 24-well plates will be stimulated using the baseline or fed serum (rom blood samples at 15, 30, 60, and 90 minutes after supplementation). The degree of mTORC1 activation will be determined as the difference in slopes between the baseline and fed sera.

  4. mTORC1 activity

    Time frame: Baseline (0 hour) to 90 minutes after supplementation

    To measure the ability of a protein source (whey protein, IPC80 or Whole Buffalo Powder) to activate mTORC1, stably transfected C2C12 muscle cells with a plasmid (pcDNA3-TOP luciferase) where the luciferase mRNA contains a 5'TOP. 5'TOP mRNA , which specifically regulated by mTORC1 activity, will be used. Differentiated C2C12TOPLuc muscle cells in 24-well plates will be stimulated using the baseline or fed serum (rom blood samples at 15, 30, 60, and 90 minutes after supplementation). The degree of mTORC1 activation will be determined as the difference in slopes between the baseline and fed sera.

Sponsors and collaborators

Lead sponsor

University of California, Davis

Other

Registry information

Official study title

The Ability of Insect-derived Protein Supplements (IPC80 and Whole Buffalo Powder, Ynsect®) to Activate mTORC1 and Protein Synthesis in Muscle

Important dates

Study start
2022
Primary completion
2023
Study completion
2024
First posted
Jul 13, 2022
Registry last updated
May 8, 2026

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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