Breast Cancer

Breast cancer is the most common cancer in women and the second most common cancer overall. Triple Negative Breast Cancer (TNBC) is a relative uncommon subtype (10-15%) of breast cancer, being characterized by poor prognosis and high metastatic potential. Currently, due to the specific TNBC molecular asset, the only available therapeutic option is the combined chemotherapy and immunotherapy as neoadjuvant regimen, followed by surgery. However, patient pathological response to neoadjuvant therapies is highly variable and no predictive biomarkers are available, yet.

 

Spatial transcriptomics of TNBC to predict patient neoadjuvant response

Molecular Oncology Unit: Dr. Ciarrocchi, Dr. Donati. Dr. Reggiani

The understanding of the contribution of stromal and immune cells to tumor biology is challenged by the technical difficulty of separating and profiling stromal cell populations from tumor cells. To overcome this problem, we take advantage of the innovative nCounter GeoMix-Digital Spatial Profiler (DSP) technology, in collaboration with the Pathology Unit. This technology allows to integrate deep profiling with precise tissue morphology evaluation. We employ this approach to conduct gene expression profiling in immune and stromal cells infiltrating TNBC biopsies to identify molecular features associated with pathological response to neoadjuvant therapy and resistance in non-responding tumors.

 

Establishing of a TNBC 3D organoids-based platform to investigate chemotherapy and immunotherapy response

Molecular Oncology Unit: Dr. Reggiani, Dr. Manzotti.

The study of tumor heterogeneity with the purpose to identify innovative and personalized therapeutic approaches requires suitable ex vivo models. To this aim, tumor specimens are collected from TNBC patients in cooperation with the Oncology Unit, Breast Surgery Unit, Pathology Unit and our Institutional BioBank. Patient-derived organoids are established and in vitro cultured, validated through immunohistochemistry and NGS profiling and matched with patient tumor characteristics. The organoid-based platform can be applied to dissect tumor biological specific features, perform drug screening and set up co-cultures with immune cells to decipher the complexity of tumor microenvironment.

 

Transcriptional reprogramming of Natural Killer (NK) cells in the treatment of TNBC

Molecular Oncology Unit: Dr. Reggiani PI (Principal Investigator), Dr. Redavid 

The implementation of neodjuvant treatment with immunotherapy has drastically improved the therapeutic outcome of TNBC patients. Among different immune cells that may be involved in anti-tumor response, Natural Killer (NK) cells appeared to be most the promising ones.

The transcriptional plasticity of NK cells is essential to modulate their tissue residency, tumor homing and effector functions, including anti-tumor cytotoxicity. The reprogramming of specific transcription factors may constitute a novel tool to enhance NK anti-tumor properties in immune cell therapies and ameliorate tumor sensitivity to neoadjuvant treatment. To this aim, the state-of-the-art approaches in the study of functional genomics and tumor microenvironment (Live-cell imaging system, flow cytometry-based functional assays and single cell RNA sequencing) will be applied to characterize the molecular pathways driving NK activation in TNBC.

 

Metabolic and epigenetic alterations in breast cancer

Molecular Oncology Unit: Dr. Pistoni, Dr. Rossi.

Abnormalities of cancer cell proliferation require important metabolic changes to support cancer initiation and progression. Thus, our goal is to understand which lipid and energetic metabolic alterations of TNBC are part of the fundamental processes of cancer initiation and progression. We aim to find metabolic fragilities in vivo and/or in vitro to make cancer cells more responsive to therapies and reduce resistance. To do so, we follow these routes:

• The usage of specific drugs, whose anti-tumor effect is known, to inhibit the transcriptional factor BRD4 (through BETi-BET inhibitors employment); cancer cells are highly dependent on the transcriptional activity of this factor. Here, we aim to demonstrate how BETi treatment modifies mitochondrial dynamism and glycolytic metabolism leading to the identification of new promising therapeutic targets through in vitro approaches.
• To comprehend the function of ATGL and lipid droplets (LDs) on metabolic rewiring (i.e. lipogenesis, lipolysis, b-oxidation), transcriptional regulation (i.e. signaling pathways, PPARa, PPARg, etc.), and cancer cell proliferation. TNBCs are highly dependent on lipid metabolism and LDs are intracellular organelles for lipid storage degraded by ATGL for the release of fatty acids. Through the integration of omics approaches, we aim to find the contribution of ATGL on molecular pathways driving cancer aggressiveness so far not yet explored in TNBC.

   

Cancer-associated adipocytes

Molecular Oncology Unit: Dr. Pistoni, Dr. Rossi.

Cancer-associated adipocytes (CAAs) are, among the cellular components of the tumor microenvironment, the main exogenous energy source for the survival of breast cancer cells.

Thanks to the collaboration with the Oncology, Pathological Anatomy, and Breast Surgery units of the Institute to obtain adipocytes from TNBC patients, we:

• Establish 2D and 3D CAAs culture in co-culture with TNBC cells to reconstruct the microenvironment of breast cancer. We aim to investigate how the functional synergy of CAAs with cancer cells affects lipid and energetic metabolism influencing cellular phenotype and proliferation.
• Use NGS approaches (RNA-seq and ATAC-seq) to identify whether CAAs influence the transcriptional and genomic heterogeneity of the TNBC cells, recapitulating the patients' variability. The purpose is to obtain a signature to correlate with the clinical data of the TNBC cells, recapitulating the patients' variability. The purpose is to obtain a signature to correlate with the clinical data of the TNBC patients to discover pathways that sustain the disease.