Role of HIF-1α in Skeletal Muscle Aging (HIF)

December 13, 2017 updated by: Istituto Ortopedico Galeazzi

Role of Hypoxia Inducible Factor-1α (HIF-1α) in Skeletal Muscle Aging

Deficits in skeletal muscle function exist during aging and muscular dystrophy, and suboptimal function has been related to factors such as atrophy, excessive inflammation and fibrosis. Sarcopenia is the age-related loss of skeletal muscle mass and function. It is now recognised as a major clinical problem for older people and research in the area is expanding exponentially. This interest stems from the fact that sarcopenia is both common and associated with serious health consequences in terms of frailty, disability, morbidity and mortality. The age-related loss of human skeletal muscle mass is due to a decrease in myofibre size and number with the loss of both fast and slow type myofibres, although the loss of fast myofibres tends to start earlier, at ∼70 years. Many factors influence the decrease in muscle mass. A significant contributor is an anabolic resistance of older skeletal muscle to protein nutrition as seen during immobilisation which can be ameliorated at least in part by resistance exercise and dietary supplementation. Other intensive areas of research are related to the loss of innervation and oxidative damage. Moreover, ineffective muscle regeneration underlies each condition and has been attributed to a deficit in myogenic potential of resident stem cells or satellite cells. It is now widely accepted that satellite cells, and generally adult stem cells, are normally quiescent and tend to reside in hypoxic areas of the tissue to preserve their undifferentiated state. To govern these processes, cells have developed a very complex machinery that is mainly regulated by a group of transcription factors known as hypoxia-inducible factors (HIFs). In particular, several observations support the idea that oxygen deprivation and HIF-1a may play a key role during ischemia to activate the regeneration process, which, after an initial hypoxic insult, needs to proceed under normoxia. On these bases, in this study we will investigate the role of HIF-1a in skeletal atrophy during aging.

Study Overview

Status

Unknown

Conditions

Detailed Description

Deficits in skeletal muscle function exist during aging and muscular dystrophy, and suboptimal function has been related to factors such as atrophy, excessive inflammation and fibrosis. Sarcopenia is the age-related loss of skeletal muscle mass and function. It is now recognised as a major clinical problem for older people and research in the area is expanding exponentially. This interest stems from the fact that sarcopenia is both common and associated with serious health consequences in terms of frailty, disability, morbidity and mortality. The age-related loss of human skeletal muscle mass is due to a decrease in myofibre size and number with the loss of both fast and slow type myofibres, although the loss of fast myofibres tends to start earlier, at ∼70 years. Many factors influence the decrease in muscle mass. A significant contributor is an anabolic resistance of older skeletal muscle to protein nutrition as seen during immobilisation which can be ameliorated at least in part by resistance exercise and dietary supplementation. Other intensive areas of research are related to the loss of innervation and oxidative damage. Moreover, ineffective muscle regeneration underlies each condition and has been attributed to a deficit in myogenic potential of resident stem cells or satellite cells. It is now widely accepted that satellite cells, and generally adult stem cells, are normally quiescent and tend to reside in hypoxic areas of the tissue to preserve their undifferentiated state. To govern these processes, cells have developed a very complex machinery that is mainly regulated by a group of transcription factors known as hypoxia-inducible factors (HIFs). In particular, several observations support the idea that oxygen deprivation and HIF-1a may play a key role during ischemia to activate the regeneration process, which, after an initial hypoxic insult, needs to proceed under normoxia. On these bases, in this study we will investigate the role of HIF-1a in skeletal atrophy during aging.

In particular, we will isolate satellite cells from muscular biopsies harvested from old sarcopenic patients and from young patients. The, we will measure HIF-1a levels and we will compare them.

Study Type

Observational

Enrollment (Anticipated)

16

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

18 years to 90 years (ADULT, OLDER_ADULT)

Accepts Healthy Volunteers

N/A

Genders Eligible for Study

All

Sampling Method

Non-Probability Sample

Study Population

Patients will be selected and evaluated in the primary care clinic. The eligibility criteria will be checked 2-4 weeks before surgery.

Muscular biopsies will be harvested from discarded material during surgery.

Description

Inclusion Criteria:

  • Sarcopenic patients (measured by DXA) affected by hip osteoarthritis, developmental dysplasia of the hip or hip fracture and undergoing hip replacement surgery (for the sarcopenic group)
  • Patients affected by traumatic ACL tears and undergoing ACL reconstruction surgery (for the control group)
  • Patients between 18 and 35 years old (for the control group)
  • Patients between 65 and 90 years old (for the sarcopenic group)
  • 18 ≤ Body Mass Index (BMI) ≤ 30 kg/m2

Exclusion Criteria:

  • Diseases that can affect bone or muscle metabolism
  • Pharmacological therapies that can interact with bone or muscle metabolism
  • Bone metastases
  • Bone infections
  • HIV, hepatitis B virus, hepatitis C virus or Treponema pallidum positivity
  • BMI ≥30kg/m2

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

Cohorts and Interventions

Group / Cohort
Intervention / Treatment
Sarcopenic group
Harvesting of muscular biopsies Muscular biopsies will be harvested from old sarcopenic patients undergoing hip replacement surgery
Muscular biopsies will be harvested during surgery; them satellite cells will be isolated and characterized in vitro.
Control group
Harvesting of muscular biopsies Muscular biopsies will be harvested from young patients undergoing Anterior Cruciate Ligament (ACL) reconstruction surgery
Muscular biopsies will be harvested during surgery; them satellite cells will be isolated and characterized in vitro.

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Evaluation of HIF-1a expression levels in satellite cells isolated from sarcopenic and young
Time Frame: 1 year
Western blot analysis of HIF-1a levels and of its downstream targets (Vascular Endothelial Growth Factor: VEGF, phosphoglycerate kinase: PGK, Prolylhydroxilases: PHD2)
1 year

Secondary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Evaluation of sarcopenic profile in satellite cells harvested from old sarcopenic patients
Time Frame: 6 months
Real Time polymerase chain reaction: PCR analysis of MURF1 and atrogin1 (markers of muscular aging)
6 months
Correlation of HIF-1a levels with the degree of sarcopenia in the satellite cells of old sarcopenic patients
Time Frame: 1 year
chromatin immunoprecipitation: ChiP seq analysis of HIF-1a downstream targets
1 year
Evaluation of differences in satellite cells number between young and old patients
Time Frame: 6 months
To evaluate differences in cell number two different immunohistochemical analyses will be used in order to obtain a ratio: 1. Immunofluorescence for PAX7 (satellite cell markers) and 2. BrdU will be performed and the ration between PAX7 positive cells and bromodeoxyuridine: BrdU positive cells will be calculated.
6 months
Evaluation of HIF-1a stabilization on satellite cells treated with PHDs inhibitors
Time Frame: 1 year
Prolylhydroxilases: PHDs inhibitors (FG-4592 e PLG) will be tested in vitro to analyze HIF-1a downstream targets (Vascular Endothelial Growth Factor: VEGF, phosphoglycerate kinase: PGK, Prolylhydroxilases: PHD2) by real time PCR
1 year
Evaluation of satellite cell proliferation in cells treated with PHDs inhibitors
Time Frame: 1 year
Prolylhydroxilases: PHDs inhibitors (FG-4592 e PLG) will be tested in vitro to analyze cell proliferation (RealTime Glow and CellTox kits)
1 year
Evaluation of satellite cell differentiation in cells treated with PHDs inhibitors
Time Frame: 1 year
Prolylhydroxilases: PHDs inhibitors (FG-4592 e PLG) will be tested in vitro to analyze cell differentiation (Immunofluorescence and Realt Time PCR analysis for differentiation markers: MyoD, Myogenin and Myosin Heavy Chain)
1 year

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Investigators

  • Principal Investigator: Laura Mangiavini, Dr, IRCCS Istituto Ortopedico Galeazzi

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (ANTICIPATED)

January 10, 2018

Primary Completion (ANTICIPATED)

January 10, 2019

Study Completion (ANTICIPATED)

July 26, 2020

Study Registration Dates

First Submitted

December 7, 2017

First Submitted That Met QC Criteria

December 7, 2017

First Posted (ACTUAL)

December 13, 2017

Study Record Updates

Last Update Posted (ACTUAL)

December 14, 2017

Last Update Submitted That Met QC Criteria

December 13, 2017

Last Verified

December 1, 2017

More Information

Terms related to this study

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

NO

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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