Advanced Therapies for Liver Metastases

Brief summary public domain (ClinicalTrials.gov)

Liver metastases (MTS) are the main cause of death for patients affected by colorectal carcinoma (CRC) and pancreatic ductal adenocarcinoma (PDAC), thus representing the major unmet clinical need for these malignancies. Based on preliminary and published data, the investigators hypothesize that innovative immune, gene, and cell therapy approaches might overcome the tolerogenic liver microenvironment and represent powerful therapeutic tools for liver MTS of PDAC and CRC. The investigators have therefore planned an observational clinical study to enroll distinct cohorts of patients (i.e., metastatic CRC, preneoplastic, metastatic, and non-metastatic PDAC) and finely characterize, through integrated state-of-the-art -omics, the immune and non-immune microenvironment of their primary tumor and/or liver metastases as well as correlate changes in the activation status and phenotype of peripheral blood leukocytes. Healthy volunteers will be enrolled as negative controls. The investigators aim at identifying: i) actionable tumor-associated antigens (TAAs) and local immune suppressive and regulatory pathways; ii) biological parameters for early diagnosis of relapse; iii) the effect of therapies on the shaping of anti-tumor immune responses. Data collected will be instrumental for the generation of novel advanced therapy medicinal products (ATMPs). Indeed, this protocol is part of a multi-partner translational program, supported by the AIRC 5 per Mille 2019 grant, focused on the development, validation, and implementation of clinical testing for ATMPs to ameliorate the cure of CRC and PDAC, and possibly to help the study of other solid tumors. Moreover, the systematic and long-term follow-up of enrolled patients will possibly point to early predictors of differential prognosis and patients' categories eligible for tailored therapies, including those with the novel ATMPs. In this regard, two additional substudies were incorporated into the main LiMeT protocol in July 2024 and January 2026, respectively, supported by supplementary funding: 1) the TREATLIVMETS (Treating Liver Metastasis) project, funded by the European Research Council (ERC) under the Horizon Europe research and innovation program; 2) the "Deciphering and targeting the immunological niche in PDAC" project, funded by the Fondazione Regionale per la Ricerca Biomedica (FRRB) under the "From Bed to Bench 2024" call. In the latter substudy, machine learning will be integrated with the spatial multi-omic …

Detailed description public domain (ClinicalTrials.gov)

MAIN STUDY: Advanced immune gene and cell therapies for liver metastases (LiMeT protocol) - AIRC5x1000 research program Background and rationale - CRC and PDAC are the second and fourth most common causes of cancer death, respectively. Patients affected by these cancers die of liver MTS. Conventional therapies are active in either primary tumors but fail in metastatic disease. The effectiveness of immunotherapy by immune checkpoint blockade and adoptive cell therapy with tumor-specific T cells is only partial or temporary in MTS from solid tumors, because of local immunosuppressive mechanisms. This is particularly relevant for hepatic MTS, in which a natural tolerogenic milieu may further sustain immunosuppression induced by cancer cells and infiltrating immune cells, such as tumor-associated macrophages (TAMs), myeloid-derived suppressor cells (MDSCs), neutrophils, and regulatory T or B cells. In this regard, evidence suggests that progression of primary or metastatic CRC correlates with poor immune infiltration and inflammation. Moreover, our published results in primary PDAC support a correlation between tumor progression and a strong type 2 anti-inflammatory/immunosuppressive signature. Efficacy of chemotherapy is known to rely also on reactivation and/or repolarization of tumor-targeting immune responses. Pre-surgical chemotherapy may induce beneficial effects and potentially targetable changes in the immune profile of CRC or PDAC MTS. The investigators hypothesize that a combination of: i) direct cancer cell killing by adoptively transferred tumor-specific T cells and ii) indirect control of tumor progression via reprogramming of the tumor microenvironment would control hepatic MTS of CRC and PDAC. Thus, the investigators propose a thorough characterization of the metastatic milieu in these two cancer types to rationally shape, on the basis of similarities and differences, innovative ATMPs. Experimental results obtained from biological samples will be correlated: i) with clinical data from patients at the time of enrollment and along their follow-up; and ii) for a subgroup of patients, with biochemical and immunological data from peripheral blood samples routinely collected as a part of their clinical follow-up, with the additional aim to identify novel biologic parameters and/or signatures to help the stratification of patients and the tailoring of treatments. Objectives - The investigators want to study: 1. the tumor mutational burden and the …

Arm public domain (ClinicalTrials.gov)

PDAC liver-MTS (): Adult patients with clinical /radiological diagnosis/suspicious of PDAC metastatic to the liver, with subsequent cytological/histological confirmation (stage IV disease, AJCC) from liver resection/metastasectomy or core liver biopsy.

Arm public domain (ClinicalTrials.gov)

CRC liver-MTS (): Adult patients with histologically or cytologically confirmed diagnosis of CRC metastatic to the liver with indication to surgical resection (upfront or after neoadjuvant therapy).

Arm public domain (ClinicalTrials.gov)

Primary non-MTS PDAC (): Adult patients with clinical/radiological diagnosis of primary non-metastatic PDAC, candidates for surgical resection with radical intent of the primary tumor (upfront surgery or after neoadjuvant therapy). These patients will be monitored for early diagnosis of metachronous hepatic PDAC MTS by follow up testing.

Arm public domain (ClinicalTrials.gov)

Healthy volunteers (): Negative control for the clinical study.

Arm public domain (ClinicalTrials.gov)

IPMN (): Adult patients with a diagnosis of Intraductal Papillary Mucinous Neoplasms (IPMN), which are considered precancerous and can progress to invasive cancer. Main duct IPMNs usually require surgical resection, while branch duct cases may be monitored with surveillance.

Primary outcome public domain (ClinicalTrials.gov)

Definition of tumor mutational burden, epigenetic and gene expression profile of the CRC and PDAC metastatic liver at bulk and at single cell level. CRC/PDAC MTS samples will be analyzed by: * Whole exome DNA sequencing * RNA sequencing * ATAC sequencing * Single cell RNA sequencing on sorted myelomonocytic and T cells infiltrating CRC/PDAC MTS samples. Naive vs chemotherapy treated lesions as well as primary vs MTS lesions will be compared to evaluate chemotherapy-induced modifications and the molecular evolution in the distinct tumor sites, respectively. Time frame: After liver metastasis/primary tumor resection/biopsy, usually within 12 months.

Primary outcome public domain (ClinicalTrials.gov)

Characterization of the immune landscapes of CRC and, if possible, PDAC liver MTS by high dimensional flow cytometry. Single cell suspension of CRC and PDAC MTS and paired autologous PBMCs and portal blood harvested before and after pre-operatory chemotherapy, will be analyzed by a series of parallel polychromatic high dimensional 28 color flow cytometry panels to study: * Conventional MHC-restricted Tab cells, the main anti-tumor effectors; * Innate-like T cells; * B cells; * Myelomonocytic populations involved in the stimulation or suppression of anti-tumor responses. Whenever available, matched primary vs MTS lesions will be compared to evaluate the immune landscapes in the distinct tumor sites. Time frame: For tissue specimens: immediately after tumor tissue resection/biopsy (analyses on fresh samples); for blood specimens: after blood drawing, usually within 12 months (analyses on fresh and/or thawed samples).

Primary outcome public domain (ClinicalTrials.gov)

Histological validation of the molecular results obtained in 1. and 2.. Validation of relevant markers and molecular pathways in terms of anatomical distribution, performed by polychromatic immunofluorence (IF, up to 6 colors) and immunohistochemistry (IHC, up to 4 colors) stainings on MTS CRC and PDAC biopsies, perilesional parenchyma and, whenever available, matched primary tumor samples. In addition to multiplex IF and IHC, some specimens will be characterized more deeply with advanced spatial proteomics technologies, such as GeoMx Digital Spatial Profiler (Nanostring). Time frame: After liver metastasis/primary tumor resection/biopsy, usually within 12 months.

Primary outcome public domain (ClinicalTrials.gov)

Definition of the antigenic landscape and TCR repertoire of CRC and PDAC liver MTS. Identification of tumor-reactive TCRs will be pursued through different and complementary strategies: * The most relevant inhibitory checkpoint genes/pathways expressed by MTS-TILs will be used to enrich for tumor specificities by cell sorting harvested from CRC/PDAC MTS tumors, portal, hepatic and peripheral blood sample. After their activation, responding T cell cultures will be subject to bulk and single-cell TCR seq; * In samples characterized by a low percentage of MTS-TILs, single cell immune profiling will be performed in order to simultaneously determine gene expression and correctly pair chain TCR sequences from individual T cells; * The library of tumor-specific TCRs identified will be transiently expressed in reporter Jurkat cells to confirm their tumor antigen specificity; * Identification and functional validation of transmembrane molecules enriched in cancer cells as possible targets for CAR design. Time frame: After liver metastasis/primary tumor resection/biopsy, usually within 12 months.

Secondary outcome public domain (ClinicalTrials.gov)

Evaluation of the molecular and cellular composition of CRC and, if possible, PDAC liver MTS by spatial transcriptomics technologies (NICHE-seq and Visium). Time frame: After liver metastasis/primary tumor resection/biopsy, usually within 12 months.

Secondary outcome public domain (ClinicalTrials.gov)

Collection of clinical follow-up data. Time frame: CRC patients: throughout the postoperative follow-up, for a maximum of 36 months; PDAC patients: throughout the postoperative follow-up, for a maximum of 24 months.

Secondary outcome public domain (ClinicalTrials.gov)

Collection and biobanking of follow-up samples from patients with CRC and metachronous PDAC MTS to the liver. Time frame: CRC: during the postoperative follow-up (up to 36 months), at six-month intervals; PDAC: during the postoperative follow-up (up to 24-months), at time of liver recurrence.

Secondary outcome public domain (ClinicalTrials.gov)

Biobanking of biospecimens collected from CRC and PDAC patients and from healthy donors. Time frame: Throughout the protocol (7 years).

Eligibility public domain (ClinicalTrials.gov)

Inclusion and exclusion criteria - CRC patients Inclusion criteria: 1. Patients with histologically or cytologically confirmed diagnosis of CRC metastatic to the liver (stage IV disease, AJCC) 2. Patients with indication to surgical resection and/or chemotherapy treatment 3. Age ≥18 4. ECOG PS 0-1 at enrollment 5. Written informed consent 6. Patients will be treated in IRCCS San Raffaele Exclusion criteria: 1. Pregnancy or lactation 2. Inability to provide a written informed consent 3. Extraepatic disease with the exception of selected cases in which the coexistence of extrahepatic disease does not constitute an exclusion criterion for hepatic resective surgery (for example in patients with extraepatic lesions in remission or in any case stabilized by chemotherapy) 4. Severe comorbidities (e.g. cardiac diseases, history of psychiatric disabilities, HIV, autoimmune disorders) 5. Concurrent or previous other malignancy within 5 years of study entry, except cured basal or squamous cell skin cancer, superficial bladder cancer, prostate intraepithelial neoplasm, carcinoma in-situ of the cervix, or other noninvasive or indolent malignancy 6. Other conditions (medical or psychiatric) that in the judgment of Investigators would make the patient an inappropriate candidate for the study Inclusion and exclusion criteria - PDAC patients Inclusion criteria: 1. Patients with clinical/radiological diagnosis/suspicious of pancreatic adenocarcinoma metastatic to the liver, with subsequent cytological/histological confirmation (stage IV disease, AJCC) 2. Age ≥18 3. Karnofsky performance status ≥50 4. Metastatic pancreatic adenocarcinoma patients with histological specimens from whole liver metastasis biopsy or core liver biopsy collected at IRCCS San Raffaele and stored in the institutional biobank Centro Risorse Biologiche (CRB-OSR) 5. Written Informed consent 6. Patients with clinical/radiological diagnosis of not metastatic primary PDAC that will undergo pancreatic resection at IRCCS San Raffaele Exclusion criteria: 1. Severe comorbidities (e.g., cardiac diseases, history of psychiatric disabilities) representing an absolute contraindication for whole or core liver metastasis biopsy 2. Pregnancy or lactation 3. Inability to provide a written informed consent 4. Metastatic pancreatic adenocarcinoma patients enrolled in other research trials entailing the analysis of the liver metastasis histological sample

Enrollment public domain (ClinicalTrials.gov)

Enrollment: 625 participants (estimated).