LEARNING OUTCOMES

The course is aimed at exploring specific example pathologies as consequences of molecular alterations, with particular focus on tumors, and at illustrating novel methodological strategies to investigate disease pathogenetic mechanisms and identify new therapeutic targets.

AIMS AND LEARNING OUTCOMES

Aim of the course is: 1) to give information about selected pathologies including tumors, immunodeficiences and cystic fibrosis as examples of diseases related to molecular alterations, with particular focus on the pathogenetic mechanisms; 2) to elucidate different treatment strategies on the basis of the disease’s molecular characteristics.

At the end of the course, students will have gained deeper knowledge of the molecular bases of representative human pathologies, and will be able to compare different therapeutic strategies by identifying pros and contras on the basis of the type of each approach and of the clinical and experimental outcomes.

PREREQUISITES

Good knowledge of cellular and molecular biology, general oncology, and immunology.

TEACHING METHODS

Oral presentations supported by Powerpoint slides.

SYLLABUS/CONTENT

General concepts of molecular pathology.

CANCER

Carcinogenesis as micro-evolutionary process. Clonal origin of tumors. Somatic mutations as the cause of tumors; genetic and epigenetic mutations. Genetic instability of tumor cells. Tumor progression: neoangiogenesis and metastases. Tumor aetiology: definition of initiators and promoters. The cancer genes: oncogenes and tumor suppressor genes. Anti-cancer therapies and multidrug resistance.

Cancer stem cells

Clonal evolution in cancerogenesis. The hierarchical model of tumor tissues: the cancer stem cell (CSC). Identification of CSC: phenotype, xenotransplantation, colony-spheroid formation, side population, and ALDH activity assays. Plasticity of the CSC model. Therapeutic strategies targeting CSC.

Tumor microenvironment

The cells of tumor microenvironment: cancer-associated fibroblasts (CAF), mesenchymal stem cells, tumor-associated macrophages (TAM) and their role in supporting tumor growth and metastasization. Caveolin-1 expression in CAF: protein function in health and disease. The “reverse Warburg effect”. Prognostic relevance of stromal Cav-1 expression in tumors.

Chronic myeloid leukemia

Molecular pathogeneis of CML: the Bcr-Abl gene. Course of disease. CML therapy: the revolution of tyrosine kinase inhibitors (TKI). Evaluation of therapy response: hematologic, cytogenetic, and molecular response. TKI resistance: Bcr-Abl mutations and leukemia stem cells (LSC). Survival pathways in LSC as new targets of CML therapies. The Programmed Cell Removal: “eat me” and “non eat me” signals; mechanisms of PCR evasion in tumor cells.

New therapeutic strategies in acute myeloid lekemia: alloreactive NK cells and CAR-T cells

Morphological and phenotypic classification of acute myeloid and lymphoid leukemias (AML, ALL). Regulation of Natural Killer (NK) cell function: activating and inhibitory receptors. Expression of NK receptor ligands on AML and ALL cells. Haploidentical hematopoietic stem cell transplantation (HSCT). NK cell allloreactivity: the KIR/HLA-class I mismatch. Correlation between NK cell alloreactivity and anti-leukemia effect: in vitro experience and clinical practice.

Adoptive immunotherapy in allo-HSCT employing genetically engineered lymphocytes: the CAR-T cells. First, second and third generation CARs. Problems of CAR-T cell therapy: solution strategies. Use of suicide genes to block side effects of CAR-T cell therapies.

PRIMARY IMMUNODEFICIENCES

Classification of primary immunodeficiences (PIDs)

Genetic alterations and pathogenetic mechanisms in PIDs: the examples of Adenosine deaminase (ADA) deficiency, X-linked Severe Combined Immune Deficiency (X-SCID), chronic granulomatous disease (CGD), and Wiskott-Aldrich Syndrome (WAS). The diverse therapeutic approaches for PIDs. Gene therapy for PIDs.

CYSTIC FIBROSIS

CFTR gene mutations and disease pathogenesis. Classes of CFTR mutations and their correlation with disease phenotype. Conventional therapies and novel therapeutic approaches for CF. The promising use of correctors and potentiators to cure CF.

RECOMMENDED READING/BIBLIOGRAPHY

Reference books:

- Alberts et al. “Biologia molecolare della cellula”,  Zanichelli ;

- Mendelsohn et al.  “The molecular basis of cancer”, Elsevier;

- Robbins e Cotran, “Le basi patologiche delle malattie”, Elsevier.

- Giacca M., “Terapia Genica”, Springer-Verlag Italia s.r.l