3D-Microscopic Muscle Architecture in Cerebral Palsy

NCT06584851 | Recruiting

• 1 other identifier: S62645
• Study Type: observational
• Target: 130
• Locations: 1 country
• Sites: 1

Brief Summary

The focus of this study is to understand and define the mechanisms of the altered muscle development and growth on a microscopic level within a long-term perspective in children with cerebral palsy and to relate these findings to muscle macroscopic properties defined by muscle imaging, to neuromuscular symptoms and to treatment. This study aims to (1) evaluate intrinsic microscopic muscle properties of young growing children with CP, and (2) to evaluate these muscle properties in relation to macroscopic properties, neuromuscular symptoms and to treatment. Improved understanding of changes in microscopic muscle properties, and how they relate to macroscopic properties and to the neuromuscular symptoms as well as how they are influenced by treatment, has the potential to delineate CP phenotypes prone to intervention and to optimize treatment protocols or develop new treatments, leading to new avenues for improving function in CP. The method to study microscopic muscle properties involves analysis of muscle biopsies (histological / immunohistochemistry analysis, SC and IC culture, gene expression). Biopsies will be collected using the minimally invasive percutaneous needle microbiopsy technique, suitable for collecting repeated samples over time in the same individual while still leading to sufficient tissue of good quality for subsequent analysis1,2. For the children with CP, the local hospital's tradition of applying general anesthesia for delivering BTX injections will be exploited to collect the muscle samples prior to the BTX session and the one-year follow-up, or general anesthesia planned for orthopedic surgery or diagnostic imaging such as MRI, etc. For the collection of the samples 3 months before the BTX session, as well as 3 months and 6 months after BTX injections, the common approach for microbiospy collection will be applied, with local sedation on the skin (Rapydan©) and fascia (Xylocaine©) and local anesthesia by using Kalinox© (nitrous oxide in oxygen) under supervision of the University Hospital PROSA team. Biopsies of TD muscles will be collected in children with no history of neurological disorder, nor musculoskeletal problems at the level of the gastrocnemius or semitendinosus, at the time of upper limb orthopedic or trauma surgery and thus always under general anesthesia. Ultrasound guided percutaneous muscle biopsy has been performed in children (2 months-18 years)3,4 and has proven to be safe and well-tolerated. A pilot study (S61110) was conducted to confirm that the microbiopsy technique is suitable for the analysis of microscopic muscle properties and is well-tolerated in children with CP. Two specific research goals are planned, with hypotheses emerging from literature.

Conditions

Spastic Cerebral Palsy

Keywords

Spastic cerebral palsy; Microscopic muscle properties; Muscle micro-biopsies; 3D freehand ultrasound; Histology; Cell culture; CP-phenotypes

Outcome Measures

Primary Outcomes (9)

• Muscle fiber size

  Cross sectional area in µm of the fiber. Muscle fiber size will be assessed on cryosection stained with an antibody cocktail specific to laminin, myosin heavy chain (MHC)-I, MHC-IIA, MHC-IIB, MHC-embryonal and immunofluorescent detection will be done using appropriate secondary antibodies.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Muscle fibre type proportion

  For each muscle, muscle fiber proportion will be assessed on cryosection stained with an antibody cocktail specific to laminin, myosin heavy chain (MHC)-I, MHC-IIA, MHC-IIB, MHC-embryonal and immunofluorescent detection will be done using appropriate secondary antibodies.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Capillary density

  Capillaries /mm2 fiber and capillary to fiber ratio. Capillaries will be detected in-situ. Capillaries will be assessed using CD31 staining.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Satellite cell density

  Number of satellite cells/mm2. Satellite cells will be detected in-situ. Co-localization of satellite cells will be assessed using Pax7 staining in conjunction with MHC-I/laminin, DAPI and CD31. Abundance of satellite cell content (Pax7+/DAPI+) will be counted and their association with the different fiber types will be identified.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Myogenic differentiation potential by means of fusion index for SC (satellite cells)

  SCs will be isolated by Fluorescence-Activated Cell Sorting (FACs) based on the presence of surface markers CD56.Behaviour of SCs will be analysed using a cell proliferation assay and a myogenic or adipogenic differentiation assay.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Myogenic differentiation potential by means of fusion index for MABs (mesoangioblasts)

  MABs will be isolated by Fluorescence-Activated Cell Sorting (FACs) based on the presence of surface markers ALP.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Overall change in muscle belly length of the medial gastrocnemius muscle and the semitendinosus muscle

  Estimation of the muscle belly length by 3DfUS. Muscle volume will be normalized to anthropometric growth.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Overall change in muscle volume of the medial gastrocnemius muscle and the semitendinosus muscle

  Estimation of the muscle belly volume by 3DfUS. Muscle volume will be normalized to anthropometric growth.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Overall changes in muscle activation patterns

  Estimation of the muscle activation patterns using instrumented spasticity assessment.

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

Secondary Outcomes (3)

• Overall changes in collagen content

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Overall change in muscle composition of the medial gastrocnemius muscle and the semitendinosus muscle by means of echo-intensity

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

• Overall change in muscle tendon length of the medial gastrocnemius muscle and the semitendinosus muscle

  Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline

Study Arms (4)

Children with spastic cerebral palsy

Children between 2 and 9 years old.

Typical developing children

Children between 2 and 9 years old.

Adolescents and adults with HSP

Adolescents between 12 and 18 years of age and adults.

Typically developing adolescents and adults

Age- and gender- matched with the recruited HSP patients

Eligibility Criteria

• Age: 2 Years - 9 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Child (0-17)
• Sampling Method: Non-Probability Sample

Study Population

Children with spastic cerebral palsy, who have routine follow-up care at the CP reference center of the University Hospitals Leuven.

You may qualify if:

• Children (boys/girls) diagnosed with predominantly spastic type of CP
• Uni- or bilateral involvement
• Gross Motor Function Classification Scale (GMFCS) Level I-III50
• to 9 years of age
• Planned for an orthopedic intervention that requires general anesthesia (botulinum toxin injections, orthopedic surgery, or diagnostic imaging such as Magnetic Resonance Imaging \[MRI\], etc.)

You may not qualify if:

• Presence of dystonia or ataxia
• Previous surgery less than 6 months at the investigated muscles
• Severe co-morbidities (that are likely to prevent proper assessment, such as severe cognitive problems)
• TYPICALY DEVELOPING CHILDREN
• Children (boys/girls)
• to 9 years of age
• Planned for a surgical intervention that requires general anesthesia (pure pediatric upper limb orthopedic surgery or trauma surgery, or ophthalmic or ear-nose-throat surgery)
• History of neurological problems
• History of orthopedic problems at the gastrocnemius or semitendinosus
• Trauma at the level of the lower limbs
• Involvement in an elite or high-performance sporting program (Children performing sports for \> 3 5 hours/week will be excluded)
• ADOSESCENTS WITH HSP of 12-18 years old and ADULTS WITH HPS
• Adolescents (boys/girls) or adults (male/female) diagnosed with HSP, SPG3a or SPG4
• Gross Motor Function Classification Scale (GMFCS) Level I-III50
• Adolescents 12 to 18 years or adults 18-40 years of age

Sponsors & Collaborators

• Universitaire Ziekenhuizen KU Leuven: lead

Study Sites (1)

UZ Leuven

Leuven, Vlaams-Brabant, 3000, Belgium

RECRUITING

Biospecimen

Retention: SAMPLES WITH DNA

Sonographically guided percutaneous needle micro-biopsies will be collected at the Medial Gastrocnemius and the Semitendinosus. Per biopsy, four muscle samples of +/- 20 mg will be collected per muscle on one location of the muscle, and useable for both histological and cell culture analysis.

MeSH Terms

Conditions

Cerebral Palsy

Condition Hierarchy (Ancestors)

Brain Damage, Chronic; Brain Diseases; Central Nervous System Diseases; Nervous System Diseases

Study Officials

• Kaat Desloovere, prof.dr.

  Department of Rehabilitation Sciences, KU Leuven, Belgium

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Anke Andries

CONTACT

+3216376753; anke.andries@kuleuven.be

Lauraine Staut

CONTACT

lauraine.staut@kuleuven.be

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Prof. dr.

Study Record Dates

• First Submitted: May 24, 2024
• First Posted: September 5, 2024
• Study Start: May 2, 2019
• Primary Completion (Estimated): December 1, 2028
• Study Completion (Estimated): December 1, 2028
• Last Updated: May 5, 2026

Record last verified: 2026-05

Locations

BE (1)