Human Amniotic Mesenchymal Cell Secretome for Neurodegeneration and Neuroinflammation

NCT06551649 | Recruiting

• 1 other identifier: 6823
• Study Type: interventional
• Target: 60
• Locations: 1 country
• Sites: 2

Brief Summary

Neurodegenerative diseases are debilitating conditions characterized by chronic inflammation, leading to dysfunction of both the non-neuronal cellular components of the central nervous system and peripheral blood immune cells. Thus, it is crucial to develop an innovative therapeutic strategy that not only effectively contrast neurodegeneration but also aims to reduce inflammation. The overall aim of the study is to provide a preclinical in vitro demonstration of the immunomodulatory and pro-regenerative potential of the human amniotic mesenchymal stromal cell (hAMSC) secretome in counteracting neurodegeneration. This potential will be evaluated in three-dimensional in vitro models of neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS) and chronic demyelinating disease (multiple sclerosis - MS). To this end, the study includes sample collection from patients without pharmacological treatment and without medical devices. Patients diagnosed with ALS, patients diagnosed with MS, and healthy volunteers will be recruited to collect blood samples and skin biopsies. Patient-specific and control organoid platforms, mimicking cellular heterogeneity and tridimensional interactions within the central nervous system including the inflammatory compartment, will be developed to be used as a valuable tool to investigate the in vitro efficacy of the hAMSC secretome.

Conditions

Amyotrophic Lateral Sclerosis; Multiple Sclerosis

Outcome Measures

Primary Outcomes (8)

• Neurite length measurement in a three-dimensional cellular model of amyotrophic lateral sclerosis.

  Organoids from induced pluripotent stem cells (iPSCs) isolated from skin biopsies of 6 amyotrophic lateral sclerosis (ALS) patients and 6 healthy controls will be generated. Neurite degeneration will be measured in fluorescence microscopy as total βIII-TUBULIN-positive fiber length divided by the number of βIII-TUBULIN-positive cells in micrometers. Measurements will be taken from at least 5 ALS and 5 control organoids to analyze correlations and differences between groups. The significance level for statistical tests and models is set at 0.05.

  18 months

• Percentage of apoptotic cells in a three-dimensional cellular model of amyotrophic lateral sclerosis.

  Organoids from induced pluripotent stem cells (iPSCs) isolated from skin biopsies of 6 amyotrophic lateral sclerosis (ALS) patients and 6 healthy controls will be generated. Apoptosis will be assessed by the percentage of propidium iodide-positive cells, representing the proportion of dead cells relative to the total number of cells. Measurements will be taken from at least 5 ALS and 5 control organoids to analyze correlations and differences between groups. The significance level for statistical tests and models is set at 0.05.

  18 months

• Area measurement of damaged myelin in a three-dimensional cellular model of multiple sclerosis.

  Organoids from induced pluripotent stem cells (iPSCs) isolated from skin biopsies of 6 multiple sclerosis (MS) patients and 6 healthy controls will be generated. Myelin sheath damage will be assessed via immunofluorescence as the reduction in myelin marker-stained area versus total section area. Measurements will be taken from at least 5 MS and 5 control organoids to analyze correlations and differences between groups. The significance level for statistical tests and models is set at 0.05.

  18 months

• Number of inflamed microglial cells in a three-dimensional cellular model of multiple sclerosis.

  Organoids from induced pluripotent stem cells (iPSCs) isolated from skin biopsies of 6 multiple sclerosis (MS) patients and 6 healthy controls will be generated. Microglial activation will be assessed by morphological variations, classifying cells as homeostatic, ramified, hyper-ramified, or amoeboid. Inflammatory microglia increase will be defined as the proportion of activated morphology cells relative to total cell count. Inflammatory markers HLA-DR and iNOS will be evaluated in terms of CD68 and IBA1 positive cells and fluorescence intensity. Measurements will be taken from at least 5 MS and 5 control organoids to analyze correlations and differences between groups. The significance level for statistical tests and models is set at 0.05.

  18 months

• Neurite length measurement in amyotrophic lateral sclerosis organoids after treatment with human amniotic mesenchymal stromal cell secretome.

  Neurite length in amyotrophic lateral sclerosis (ALS)-derived organoids treated with human amniotic mesenchymal stromal cell (hAMSC) secretome will be quantified using fluorescence microscopy as the total βIII-TUBULIN-positive fiber length divided by the number of βIII-TUBULIN-positive cells. The ability of the hAMSC secretome to restore ALS organoids (at least 5) to values close to healthy-donors organoids (at least 5) will be evaluated. The significance level is set at 0.05.

  24 months

• Percentage of apoptotic cells in amyotrophic lateral sclerosis organoids after treatment with human amniotic mesenchymal stromal cell secretome.

  Apoptosis will be measured by the reduction in propidium iodide-positive cells, expressed as a percentage of the total cells, in amyotrophic lateral sclerosis (ALS)-derived organoids treated with human amniotic mesenchymal stromal cell (hAMSC) secretome. The ability of the hAMSC secretome to restore ALS organoids (at least 5) to values close to healthy-donors organoids (at least 5) will be evaluated. The significance level is set at 0.05.

  24 months

• Area measurement of damaged myelin in multiple sclerosis organoids after treatment with human amniotic mesenchymal stromal cell secretome.

  Reduction of demyelinated areas in multiple sclerosis (MS)-derived organoids treated with human amniotic mesenchymal stromal cell (hAMSC) secretome will be measured by the increase in mature oligodendrocyte markers relative to total section area. The ability of the hAMSC secretome to restore MS organoids (at least 5) to values close to healthy-donors organoids (at least 5) will be evaluated. The significance level is set at 0.05.

  24 months

• Number of immunoregulatory microglial cells in multiple sclerosis organoids after treatment with human amniotic mesenchymal stromal cell secretome.

  The acquisition of an immunoregulatory phenotype will be determined in multiple sclerosis (MS)-derived organoids after treatment with human amniotic mesenchymal stromal cell (hAMSC) secretome by analyzing cells that are iNOS-negative, Arginase-positive, and HLA-DR low. Immunofluorescence analysis will count the number of cells positive for inflammatory/immunoregulatory markers out of total microglial IBA1+ cells. Fluorescence intensity changes will also be evaluated. The ability of the hAMSC secretome to restore MS organoids (at least 5) to values close to healthy-donors organoids (at least 5) will be evaluated. The significance level is set at 0.05.

  24 months

Secondary Outcomes (1)

• Percentage of altered immune phenotypic markers in amyotrophic lateral sclerosis and multiple sclerosis patients

  18 months

Study Arms (3)

Patients diagnosed with amyotrophic lateral sclerosis

EXPERIMENTAL

Amyotrophic lateral sclerosis patients will be recruited to obtain venous blood draw and skin biopsy, to optimize a three-dimensional cellular model of multiple sclerosis in which determine the neuroprotective and anti-inflammatory/immunomodulatory effects of the human amniotic mesenchymal stromal cell secretome.

Other: Venous blood draw and skin biopsy

Patients diagnosed with multiple sclerosis

EXPERIMENTAL

Multiple sclerosis patients will be recruited to obtain venous blood draw and skin biopsy, to optimize a three-dimensional cellular model of multiple sclerosis in which determine the neuroprotective and anti-inflammatory/immunomodulatory effects of the human amniotic mesenchymal stromal cell secretome.

Other: Venous blood draw and skin biopsy

Healthy volunteers, patients' unaffected spouses

ACTIVE COMPARATOR

Spouses of patients will be enrolled as healthy controls, matching their age and male-to-female ratio for both conditions, to obtain venous blood draw and skin biopsy, in order to develop a control model against which to evaluate the preclinical efficacy of the human amniotic mesenchymal stromal cell secretome.

Other: Venous blood draw and skin biopsy

Interventions

Venous blood draw and skin biopsy OTHER

The skin tissue will be used to isolate fibroblasts, which will then be reprogrammed into induced pluripotent stem cells (iPSCs). These iPSCs will be differentiated to develop patient-specific and control organoids. The blood samples will be used to isolate peripheral blood mononuclear cells (PBMC) to study the contribution of inflammation in the in vitro models developed.

Healthy volunteers, patients' unaffected spouses; Patients diagnosed with amyotrophic lateral sclerosis; Patients diagnosed with multiple sclerosis

Eligibility Criteria

• Age: 20 Years - 60 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)

You may qualify if:

• For patients with amyotrophic lateral sclerosis (ALS): aged between 50 and 60 years and similar age of onset and duration of the disease.
• For patients with multiple sclerosis (MS): with recently confirmed diagnosis of MS, aged between 20 and 50 years and considering the male-to-female ratio in MS of approximately 2:1.
• For healthy volunteers: spouses of patients unaffected by any neurological disease and matching their age and gender ratio for both conditions.

You may not qualify if:

• Patients who do not consent to participate in the study.
• MS patients who have received treatment with immunomodulators or corticosteroids and are in an acute phase of the disease.

Sponsors & Collaborators

• Fondazione Policlinico Universitario Agostino Gemelli IRCCS: lead

Study Sites (2)

Fondazione Policlinico Universitario A. Gemelli IRCCS

Roma, 00168, Italy

NOT YET RECRUITING

Fondazione Policlinico Universitario Agostino Gemelli IRCCS

Rome, 00168, Italy

RECRUITING

MeSH Terms

Conditions

Amyotrophic Lateral Sclerosis; Multiple Sclerosis

Interventions

Blood Specimen Collection

Condition Hierarchy (Ancestors)

Spinal Cord Diseases; Central Nervous System Diseases; Nervous System Diseases; Motor Neuron Disease; Neurodegenerative Diseases; TDP-43 Proteinopathies; Neuromuscular Diseases; Proteostasis Deficiencies; Metabolic Diseases; Nutritional and Metabolic Diseases; Demyelinating Autoimmune Diseases, CNS; Autoimmune Diseases of the Nervous System; Demyelinating Diseases; Autoimmune Diseases; Immune System Diseases

Intervention Hierarchy (Ancestors)

Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Investigative Techniques

Study Officials

• Ornella Parolini

  Fondazione Policlinico Universitario A. Gemelli, IRCCS

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Ornella Parolini

CONTACT

+0630154464; ornella.parolini@policlinicogemelli.it

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NON RANDOMIZED
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: July 16, 2024
• First Posted: August 13, 2024
• Study Start: January 17, 2025
• Primary Completion: March 31, 2026
• Study Completion (Estimated): September 30, 2026
• Last Updated: August 14, 2025

Record last verified: 2025-08

Locations

IT (2)