Cellular and Molecular Physiology Laboratory

Fisiologia

Referent: Prof. Gerardo Biella

Co-workers: Giorgia Scarpellino (RTDa)

Molecular Cell Physiology

The role of Ca2 + signals in angiogenesis and therapeutic implications.

Recent studies have shown that the process of vasculogenesis, or the formation of de novo blood vessels starting from endothelial progenitor cells (EPC), takes place not only during embryonic development, but also in conjunction with the angiogenic switch of solid tumors. Our group has demonstrated for the first time the importance of Ca2+ signals in the angiogenic activity of human colony-forming endothelial cells (ECFC). In particular, we have shown that both VEGF and chemokine SDF-1 induce an increase in the intracellular concentration of Ca2+ ([Ca2+] i) which stimulates proliferation, migration, tubulogenesis and formation of new vessels in vivo. Our current research activity therefore aims to: 1) continue the investigation on ion channels, in particular on TRP channels and SOCE, which control the angiogenic activity of ECFCs through an increase in [Ca2+]i; 2) study the mechanism by which reactive oxygen species and the NADH oxidase enzyme contribute to supporting the pro-angiogenic oscillations of Ca2+ evoked by VEGF; 3) investigate whether and how the machinery underlying the genesis of Ca2+ signals in ECFCs is altered in patients with cardiovascular diseases. In parallel, our group is studying the possibility of inducing pro-angiogenic Ca2+ signals in ECFC by optical stimulation of photosensitive conjugated polymers. Our attention is especially directed to TRP channels sensitive to reactive oxygen species, such as TRPV1, TRPM2 and TRPA1. The ultimate goal of this line of research is the identification of an alternative strategy to stimulate the angiogenic activity of endogenous ECFCs in patients suffering from cardiovascular diseases, such as heart failure and peripheral limb disease.

Role of ion channels in cancer cells.

It has long been shown that ion channels, especially ion channels permeable to Ca2+, are involved in the genesis of the cancer hallmarks described by Hanahan and Weinberg, such as uncontrolled proliferation, resistance to apoptosis, and metastasis. The present research project aims to study the role of Ca2+ signals in proliferation and migration processes in primary cell lines established from patients affected by two different types of neoplasms, such as renal cell carcinoma and colorectal carcinoma. Specifically, our attention is focused on the Ca2+ toolkit of metastatic cells of hepatic origin, with a particular interest in SOCE, TRP channels and lysosomal channels (TPC and TRPML1). This approach aims to identify a battery of Ca2 + permeable ion channels that can be pharmacologically targeted for therapeutic purposes. Recently, we have begun to investigate the remodeling of Ca2+ signals in cytotoxic T lymphocytes and tumor-infiltrating T lymphocytes isolated from liver metastases of patients with colorectal cancer. Again, the project aims to improve the effectiveness of immunotherapy by activating or inhibiting specific ion channels involved in cytotoxic activity (eg SOCE).

Neurophysiology

Role of endothelial ion channels in neurovascular coupling.

Neurovascular coupling is the mechanism by which cerebral blood flow increases or decreases in response to corresponding changes in neural activity. The most accredited hypothesis attributes exclusively to neurons and astrocytes the ability to release vasoactive mediators in response to synaptic activity. This research project is investigating for the first time the ability of the cerebral endothelium to directly detect the synaptic release of neurotransmitters. Our attention is focusing on the sensitivity of hCMEC/D3 cells, a widely validated model for the study of the human brain endothelium, to the neurotransmitters glutamate and acetylcholine. Specifically, the present project aims to: 1) study the mechanisms responsible for the genesis of the intracellular Ca2+ signals that arise following stimulation with these neurotransmitters; 2) check whether and how these Ca2+ signals induce the synthesis of nitrogen monoxide (NO), the main vasodilating agent of the brain; 3) investigate whether NO released by brain endothelial cells can act not only on vascular tone, but also on synaptic activity. Recently, our research activity has extended to the study of membrane ion channels that regulate the activity of hCMEC/D3 cells independently of synaptic stimulation. Using the patch-clamp technique, we are investigating the expression and biophysical characteristics of some voltage-dependent channels, Cl- channels and different TRP channels (eg. TRPV1 and TRPA1).

The role of Ca2+ signals in angiogenesis and therapeutic implications:

  • F. Moccia, UNIMOL;
  • G. Guidetti, DBB, UNIPV;
  • U. Laforenza e K. Lefkimmiatis, Dipartimento di Medicina Molecolare, UNIPV;
  • M.R. Antognazza, Istituto Italiano di Tecnologia - Milano;
  • E. Di Pasquale, IRGB-CNR, Rozzano;
  • F. Lodola, Università di Milano-Bicocca.

Role of ion channels in cancer cells:

  • F. Moccia, UNIMOL;
  • G. Guidetti, DBB, UNIPV;
  • U. Laforenza e K. Lefkimmiatis, Dipartimento di Medicina Molecolare, UNIPV;
  • D. Montagna, P. Pedrazzoli, IRCCS Policlinico S. Matteo, Pavia.

Role of endothelial ion channels in neurovascular coupling:

  • F. Moccia, UNIMOL;
  • E. D’Angelo, Dipartimento di Scienze del Sistema Nervoso e del Comportamento, UNIPV;
  • G. Sancini, Università di Milano-Bicocca;
  • D. Lim, Università del Piemonte Orientale;
  • R. Berra-Romani, Benemérita Universidad Autónoma de Puebla, Mexico;
  • S. Dragoni, University College London, UK