Metastatic Colorectal Cancer
Conditions
Keywords
Circulating extracellular vesicles (EVs), Biomarker discovery
Brief summary
Metastatic colorectal cancer (mCRC) remains a major clinical challenge due to heterogeneous treatment responses and the development of therapeutic resistance. Current tissue-based molecular profiling is limited by invasiveness and the inability to enable longitudinal monitoring. Extracellular vesicles (EVs) represent a promising source of non-invasive biomarkers, carrying stable RNA and proteins reflective of tumor biology. This study aims to identify and validate EV-derived transcriptomic and proteomic biomarkers to monitor disease evolution, predict treatment response, and support personalized management of patients with mCRC.
Detailed description
Observational, ambispective, multicenter, non-interventional study without drugs or medical devices, aimed at the identification, validation, and clinical evaluation of extracellular vesicle (EV)-derived biomarkers in patients with metastatic colorectal cancer (mCRC). The study will include patients diagnosed with metastatic colorectal cancer (mCRC), divided into a retrospective cohort and a prospective cohort. This study aims to identify and validate circulating extracellular vesicle (EV)-derived biomarkers, including EV-RNA and EV-protein (EV-PROT) profiles, in patients with metastatic colorectal cancer (mCRC). The goal is to develop an integrated non-invasive biomarker panel to support patient stratification, treatment response prediction, and longitudinal disease monitoring. The study will be conducted through three main phases: Discovery phase: identification of novel EV-RNA and EV-PROT biomarkers from plasma samples of mCRC patients, integrated with spatial transcriptomic and proteomic analyses of tumor tissues and functional evaluation in patient-derived colorectal cancer organoids (CRC-org). Retrospective validation: validation of selected biomarkers in an independent mCRC cohort using droplet digital PCR (ddPCR) for EV-RNA and nano-flow cytometry for EV-PROT, combined with clinical outcome analyses and functional studies using CRC-org-derived 3D tumor models (assembloids). Prospective validation: assessment of biomarker clinical value in an independent prospective mCRC cohort through longitudinal plasma analyses at different treatment time points, evaluating associations with treatment response, progression-free survival (PFS), and overall survival (OS). The integration of molecular, clinical, and experimental data will support the biological characterization of EV-derived biomarkers and their potential role in tumor progression, therapeutic response, and resistance mechanisms. The statistical analysis will be conducted to evaluate the association between EV-RNA and EV-PROT biomarkers and clinical disease outcomes. Data will be analyzed using descriptive methods and appropriate statistical tests to compare groups and assess changes over time.
Interventions
OTHERBlood sample
Collection, in conjunction with blood sampling performed as part of routine clinical practice, of a 9 mL blood sample in an EDTA tube at the following time points: before treatment initiation, at the first radiological reassessment, and at disease progression. This sample collection is part of the standard clinical management of patients with metastatic colorectal cancer (mCRC).
Sponsors
Fondazione Policlinico Universitario Agostino Gemelli IRCCS
Study design
Observational model
COHORT
Time perspective
OTHER
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
No
Inclusion criteria
Retrospective Cohort Inclusion Criteria: * Age ≥ 18 years; * Histologically confirmed diagnosis of colorectal adenocarcinoma; * Radiological evidence of metastatic or locally advanced unresectable disease; * First-line treatment for metastatic disease already completed or ongoing according to standard clinical practice; * Availability of stored biological samples collected at least at one of the protocol-defined time points (baseline, post-treatment, disease progression); * Informed consent acquisition will be performed in accordance with Article 110-bis of the Italian Privacy Code. Retrospective Cohort
Exclusion criteria
* Absence of suitable biological samples or samples unsuitable for molecular analyses; * Incomplete clinical data preventing the performance of the planned analyses; * Insufficient quality of stored biological samples for the assessment of EV-RNA and EV-PROT analyses. Prospective Cohort Inclusion Criteria: * Age ≥ 18 years; * Histologically confirmed diagnosis of colorectal adenocarcinoma; * Radiological evidence of unresectable metastatic disease; * Patients not previously treated for metastatic disease (prior completed adjuvant/neoadjuvant therapy is allowed if completed ≥ 6 months before study enrollment); * Indication for first-line treatment with chemotherapy ± biological therapy according to standard clinical practice; * Ability to understand the study procedures and provide written informed consent. Prospective Cohort
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Identification and Validation of Extracellular Vesicle-Derived Biomarkers in Metastatic Colorectal Cancer | 3 years | The study aims to identify and validate circulating extracellular vesicle (EV)-derived biomarkers, including EV-RNA and EV-protein (EV-PROT) signatures, in patients with metastatic colorectal cancer (mCRC). The objective is to develop and validate an integrated, non-invasive biomarker panel to support patient stratification, predict response to systemic therapies, and enable longitudinal monitoring of disease evolution throughout treatment. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Identification of EV-RNA and EV-PROT Biomarkers in Metastatic Colorectal Cancer (Discovery Phase) | 3 years | The discovery phase aims to identify novel EV-RNA and EV-PROT biomarkers derived from circulating extracellular vesicles in patients with metastatic colorectal cancer (mCRC). Biomarker candidates identified in plasma samples will be integrated with spatial transcriptomic and proteomic analyses of primary tumor tissues to assess their biological relevance. Functional validation will be performed using patient-derived colorectal cancer organoids (CRC-org), evaluating the effects of chemotherapy and targeted therapies on EV-derived biomarker expression. |
| Retrospective Validation of Biomarker Candidates in an Independent Cohort and Experimental Models | 3 years | The identified EV-RNA and EV-PROT biomarkers will be retrospectively validated in an independent cohort of patients with metastatic colorectal cancer (mCRC). EV-RNA biomarkers will be assessed by droplet digital PCR (ddPCR), while EV-PROT biomarkers will be validated through nano-flow cytometry. Biomarker data will be integrated with clinical and pathological parameters to evaluate their association with treatment response, disease progression, and survival outcomes. Functional validation will be performed using patient-derived colorectal cancer stem cell models (CRC-org). |
| Prospective Validation and Assessment of the Clinical Utility of Biomarkers | 3 years | The final phase will prospectively validate EV-RNA and EV-PROT biomarkers in an independent cohort of patients with metastatic colorectal cancer (mCRC) enrolled at two participating centers. Longitudinal plasma sample analyses collected at baseline, first radiological evaluation, and disease progression will assess the association between biomarker levels and clinical outcomes, including treatment response, progression-free survival (PFS), and overall survival (OS). Data from CRC-org models and 3D assembloids will be integrated with clinical and molecular findings to support the biological interpretation of the identified biomarkers and their potential role in therapeutic response and resistance mechanisms. |
Contacts
CONTACTMaria Alessandra Calegari, MD
STUDY_DIRECTORDonatella Lucchetti, PhD
Catholic University of the Sacred Heart
Outcome results
None listed