Biology subjects

Zagnoni, M.

Publications and source records attributed to Zagnoni, M..

3 recordsLinked to original sources

bioRxiv2021

Assessment of CAR-T mediated and targeted cytotoxicity in 3D microfluidic TBNC co-culture models for combination therapy

Chimeric antigen receptor (CAR)-T cell therapy is efficacious against many hematological malignancies; however, their therapeutic application to treat solid tumors presents further challenges. A better understanding of how the solid tumor microenvironment (TME) impacts CAR-T anti-tumor effects would enable the selection of effective combination therapies to decipher the optimal course of treatment for patients and to better engineer CAR-Ts. Classical 2D in vitro models do not provide sufficient recapitulation of the native human TME, and in vivo models, such as patient-derived xenografts, are costly, complex and labor intensive. Here, we present a novel 3D, miniaturized assay for the evaluation of EGFR-targeted CAR-T cell cytotoxicity and specificity on tumor-stroma triple-negative breast cancer models in microfluidic devices. CAR-T cells were shown to home towards EGFR-expressing cancer cells to elicit a cytotoxic effect, whilst leaving low EGFR-expressing fibroblasts viable, an effect which was enhanced through combination anti-PD-L1 therapy and carboplatin chemotherapy. Hence, we propose this proof-of-concept immunoassay as a future preclinical screening tool for the development of novel immunotherapeutics and for use in personalized medicine.

bioengineering↗

bioRxiv2020

Metabolic control of tumour extracellular matrix production in cancer-associated fibroblasts

TElevated production of collagen-rich extracellular matrix (ECM) is a hallmark of cancer associated fibroblasts (CAFs) and a central driver of cancer aggressiveness. How to target ECM production to oppose cancer is yet unclear, since targeting CAFs has been shown to restrain but also promote cancer progression. Metabolic rewiring is a hallmark of CAFs. Here we find that proline, which is a highly abundant amino acid in collagen proteins, is newly synthesised from glutamine to make tumour collagen in breast cancer xenografts, and that its production is elevated in breast cancer CAFs. PYCR1 is the rate-limiting enzyme for proline synthesis and is highly expressed in the tumour stroma of breast cancer patients and in CAFs. Reducing PYCR1 levels in CAFs is sufficient to reduce tumour collagen production, tumour growth and metastatic spread in vivo and cancer cell proliferation in vitro. PYCR1 and COL1A1 are overexpressed in patients with invasive ductal carcinoma with poor prognosis. Both collagen and glutamine-derived proline synthesis in CAFs are enhanced by increased pyruvate dehydrogenase-derived acetyl-CoA levels, via gene expression regulation through the epigenetic regulator histone acetyl transferase EP300. Altogether, our work unveils unprecedented roles of CAF metabolism to support pro-tumorigenic collagen production. PYCR1 is a recognised cancer cell vulnerability and potential target for therapy, hence, our work provides evidence that targeting PYCR1 in tumours may have the additional benefit of halting the production of pro-tumorigenic ECM.

cancer biology↗

bioRxiv2019

Seeing around corners: Cells create chemotactic gradients to solve mazes and respond to distant cues in complex environments.

Chemotaxis, in which cells steer using chemical gradients, drives fundamental biological processes like embryogenesis, metastasis and immune responses. Self-generated chemotaxis, where cells break down abundant attractants to create gradients, is an important but under-studied aspect of physiological navigation. Here we show that self-generated gradients allow cells to navigate arbitrarily complex paths and, remarkably, make accurate choices about pathways they have not yet encountered. This enables cells to solve microfluidic mazes, even with initially homogeneous environments and distant correct destinations. We combine computational models and experiments to understand how cells anticipate environmental features, and how decision accuracy is determined by path complexity, attractant diffusibility and cell speed. This permits mazes that are easy or hard for cells to resolve, despite similar appearances. Counterintuitively, slowly-diffusing attractants can generate a "mirage", making cells prefer dead ends over correct paths. In vivo environments resemble complex mazes, and only self-generated gradients realistically explain cell behaviour.

cell biology↗