Discovery and Validation of Protein Structural Complexes in Circulating Biofluids As Novel Biomarkers for Early Diagnosis, Prognosis and Therapeutic Management of Patients Affected by Neurodegenerative Disorders

NCT06803784 | Recruiting DMC

• 1 other identifier: PNRR-MCNT2-2023-12377375
• Study Type: observational
• Target: 110
• Locations: 1 country
• Sites: 1

Brief Summary

Neurodegenerative disorders (NDDs), such as Parkinson¿s disease (PD), Alzheimer¿s disease (AD), Frontotemporal dementia (FTD) and Amyotrophic Lateral Sclerosis (ALS) are characterized by aggregation and intracellular accumulation of misfolded proteins, which are believed to play a key role in synaptic dysfunction and neuronal death. Protein structural complexes in biofluids have been proposed to mirror pathological conditions suggesting their use as biomarkers for NDDs characterized by protein aggregation. In this framework, we plan to: i) collect a large cohort of NDD and prodromal patients and healthy subjects using standardized clinical and genetics procedures; ii) apply a novel method based on genomics, proteomics and bioinformatic analysis to map protein complexes in biofluids; iii) identify novel circulating biomarkers and correlate them to genetic profiling and disease endophenotypes, and; iv) validate the biological properties in human brain tissue and dopaminergic cultures.

Conditions

Parkinson Disease; Amyotrophic Lateral Sclerosis (ALS); Frontotemporal Dementia (FTD); Alzheimer's Disease (AD)

Keywords

protein structural complexes in Neurodegenerative diseases; circulating biomarkers for neurodegenerative diseases

Outcome Measures

Primary Outcomes (10)

• motor symptoms of PD patients will be evaluated with Hoehn and Yahr (HY) score

  The Hoehn and Yahr Scale will be used to measure how Parkinson's symptoms progress and the level of disability. Itincludes stages 1 through 5: Stage 1. Unilateral involvement only, usually with minimal or no functional impairment; Stage 2 = Bilateral disease, without impairment of balance; Stage 3 = Mild to moderate bilateral disease; some postural instability; physically independent. Stage 4 = Severe disability; still able to walk or stand unassisted. Stage 5 = Wheelchair bound or bedridden unless aided.

  2 years

• motor and non motor symptoms of PD patients will be evaluated with MDS-UPDRS

  The MDS-UPDRS has four parts, namely, I: Non-motor Experiences of Daily Living; II: Motor Experiences of DailyLiving; III: Motor Examination; IV: Motor Complications. Twenty questions are completed by the patient/caregiver

  2 years

• clinical evaluation of cognitive impairment of PD, AD, ALS/FTD patients

  PD patients will be evaluated with the Montreal Cognitive Assessment (MoCA) test. This is a test used by healthcare providers to evaluate people with memory loss or other symptoms of cognitive decline. The MoCA contains 30 questions and checks different types of cognitive or thinking abilities. These include:orientation, short-term memory/delayed recall, executive function/visuospatial ability, clock-drawing test.- assessment of language disorders. ALS/FTD - assessment of cognitive disorders (and classification according to Strong criteria, 2017); - assessment of language disorders. AD patients: - assessment of cognitive disorders (MMSE, MOCA test, clock test); \- assessment of language disorders.

  2 years

• clinical evaluation of ALS/FTD patients

  Clinical classification according to El Escorial - revised

  2 years

• identification of variants/mutations

  PD patients: the number of multiple rare (Minor allele frequency, MAF\<0.01) deleterious (missense, non sense, frameshift and splicing) variants in PD genes will be counted in PD patients and unrelated healthy subjects. Case-control assoiciation study will evaluate the PD risk. AD, ALS/FTD: deleterious variants (missense, non sense, frameshift and splicing) in responsible genes will be considered.

  2 years

• Identification of protein complexes in CSF

  Protein complexes present in the CSF of PD/AD/FTD/ALS patients and control subjects recruited in the two Clinical Centers participating in the study will be analyzed by size exclusion chromatography (SEC) and by label-free proteomic analysis. The complexes will be separated and fractionated by SEC and identified and quantified by mass spectrometers. Statistical analysis will allow to identify the presence/absence or the different abundance of protein complexes in patients compared to controls and therefore the identification of biomarkers and potential therapeutic targets.

  2 years

• Bioinformatic analysis for the identification of proteins and molecular pathways involved

  Through advanced bioinformatics analysis, multivariate statistical analysis techniques and machine learning, proteins and molecular pathways involved in different neurodegenerative diseases and their development will be identified. This will allow us to understand and improve the knowledge of these pathologies and to identify potential therapeutic or diagnostic targets/pathways.

  2 years

• Validation of protein complexes in plasma

  The key marker protein complexes identified in Phase 1 will be validated in plasma samples from PD/AD/FTD/ALS patients and control subjects through targeted mass spectrometry analyses and through immunochemical technical analyses such as ELISA, Western blot and immunoassays.

  2 years

• Association analysis with endophenotypes

  Correlation analyses will be aimed at evaluating whether the presence of aggregates impacts on the age of onset (AAO), neurological symptoms, non-motor symptoms, as well as the response to L-dopa treatment. In addition, genetic data from PD patients will be integrated with biomolecular data to identify altered molecular pathways. ROC curves will be used to evaluate the diagnostic power of the test.

  2 years

• Analysis of the biological impact of protein complexes identified in CSF on the viability of mdDA dopaminergic neurons obtained from iPSCs of patients and controls

  Patient and control iPSC lines will be differentiated into mesodiencephalic dopaminergic neurons using published protocols. Cells will be exposed to protein aggregates purified from patient CSF and the cells' vulnerability to neurodegeneration will be assessed.

  2 years

Interventions

Genetic: whole genome sequencing GENETIC

Genetic: whole genome sequencing PD Partecipants will be assessed for disease progression: Stadio di Hoehn and Yahr, MDS-UPDRS part III, MOCA test, no motor symptoms, therapy and LID occurrence, Sleep disorders. Partecipants will be subjected to peripheral blood sampling for the purification of DNA, plasma, serum, PBMC and generation of hiPSC. DNA of each partecipants we will analysed by whole genome sequencing by next generation sequencing to identify any variant in candidate PD genes

whole exome sequencing GENETIC

Genetic: whole genome sequencing AD Partecipants will be assessed for disease progression: * assessment of cognitive disorders (MMSE, MOCA test, clock test); * assessment of language disorders; * current drug therapy (and possible start date of treatment); * date of onset of cognitive disorders; * acquisition and assessment of imaging data where present (MRI, CT, PET). DNA of each partecipants we will analysed by whole genome sequencing by next generation sequencing to identify any variant in candidate AD genes

Eligibility Criteria

• Age: 20 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

The study will be performed on a total population of 510 subjects (140 PD, 150 AD/ALS/FTD, 120 controls, 100 relatives of patients carrying mutations), of these, 90 patients with a clinical diagnosis of neurodegenerative disease (40 PD cases, 30 AD cases, 20 ALS/FTD cases) will be recruited in this study. The study also includes the recruitment and analysis of 20 control individuals selected among those who will undergo CSF sampling in a diagnostic regimen for non-inflammatory and non-neurodegenerative diseases. The validation cohort, partially already available at the Clinical Centers involved in the study, is composed of 400 subjects of which: 100 PD patients (UO1), 100 AD/FTD/ALS patients (UO2), 100 healthy family members of NDD patients carrying pathogenic mutations and who have not yet developed the disease (UO1) and 100 control individuals (UO1).

You may qualify if:

• For the IRCCS INM Neuromed, patients will be recruited from those affiliated with the Center for the Study and Treatment of Parkinson's Disease of the Neuromed Institute of Pozzilli. Affected subjects will be selected according to the criteria proposed by Gelb et al in 1999. This is a very pragmatic scheme based on the presence of four cardinal signs, the response to a test administration of Levodopa and the absence of atypical signs:
• A) Presence of at least 2 of the 4 cardinal signs (tremor, rigidity, bradykinesia, asymmetric onset) one of which must be tremor or bradykinesia; B) Absence of atypical symptoms such as: i) early postural instability, freezing phenomena, cognitive deterioration, hallucinations, pathological involuntary movements, vertical gaze paralysis; ii) proven causes of secondary parkinsonism (focal lesions, drugs, toxic substances); C) Documented response to the use of L-dopa or dapamine agonists (or lack of an adequate therapeutic attempt with L-dopa or dopamine agonists).

You may not qualify if:

• PD PATIENTS
• pre-existing psychiatric pathologies;
• neurodegenerative neurological diseases such as multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's, neuromuscular diseases, epilepsy;
• diagnosis of dementia;
• AD/FTD/ALS PATIENTS
• pre-existing psychiatric pathologies;
• previous diagnosis of other neurodegenerative neurological diseases;
• patients unable to sign informed consent.
• CONTROLS
• pre-existing psychiatric pathologies;
• neurodegenerative neurological diseases such as Parkinson's, multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's, neuromuscular diseases, epilepsy;
• diagnosis of dementia;
• depression;
• prolonged intake of anxiolytic, antidepressant, antipsychotic, sleep-inducing, cognitive stimulant drugs.

Sponsors & Collaborators

• Neuromed IRCCS: lead
• Università del Piemonte Orientale AOU Maggiore della Carità - Novara: collaborator

Study Sites (1)

IRCCS INM Neuromed

Pozzilli, Italy, 86077, Italy

RECRUITING

Biospecimen

Retention: SAMPLES WITH DNA

1. blood samples for purification of DNA, plasma, serum and PBMC 2. Cerebrospinal fluid

MeSH Terms

Conditions

Parkinson Disease; Amyotrophic Lateral Sclerosis; Frontotemporal Dementia

Interventions

Exome Sequencing

Condition Hierarchy (Ancestors)

Parkinsonian Disorders; Basal Ganglia Diseases; Brain Diseases; Central Nervous System Diseases; Nervous System Diseases; Movement Disorders; Synucleinopathies; Neurodegenerative Diseases; Spinal Cord Diseases; Motor Neuron Disease; TDP-43 Proteinopathies; Neuromuscular Diseases; Proteostasis Deficiencies; Metabolic Diseases; Nutritional and Metabolic Diseases; Frontotemporal Lobar Degeneration; Dementia; Neurocognitive Disorders; Mental Disorders

Intervention Hierarchy (Ancestors)

Whole Genome Sequencing; Sequence Analysis, DNA; Sequence Analysis; Genetic Techniques; Investigative Techniques

Study Officials

• TERESA ESPOSITO, PhD

  IRCCS INM Neuromed

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Teresa Esposito, PhD

CONTACT

+39 0865915249; teresa.esposito@igb.cnr.it

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Researcher head of the CNR lab

Study Record Dates

• First Submitted: January 27, 2025
• First Posted: January 31, 2025
• Study Start: February 4, 2025
• Primary Completion (Estimated): August 31, 2026
• Study Completion (Estimated): December 1, 2026
• Last Updated: March 18, 2025

Record last verified: 2025-03

Data Sharing

• IPD Sharing: Will not share

Locations

IT (1)