RESEARCH ACTIVITY

Two main projects of the Indraccolo lab will be presented.

METABOLIC ADAPTATION OF TUMORS TO ANTI-ANGIOGENIC THERAPY

The focus of our research is to investigate the biology of metabolic heterogeneity of tumors and how this heterogeneity modulates and is itself modulated by anti-angiogenic therapy. In this conceptual framework, we are also keen to investigate the possible impact on tumor angiogenesis of therapeutic strategies targeting glycolysis and their combination with anti-angiogenic therapy.

Background

Albeit less studied compared with genetic heterogeneity, it is increasingly recognized that tumors are metabolically heterogeneous. In general, both inter-tumor and intra-tumor metabolic heterogeneity can be observed and the biological basis of this phenomenon remains largely unexplored. Enhanced glycolytic activity, one of the best known metabolic hallmarks of cancer, is an heterogeneous trait of tumors and is inter-connected with another hallmark of cancer, namely angiogenesis. Indeed, it has been shown that soluble factors released by highly glycolytic tumor cells, such as lactic acid, modulate the stromal microenvironment, contributing to promote angiogenesis. On the other hand, it has also been observed that endothelial cells preferentially use glycolysis as energy source, especially during angiogenesis. Therefore, in the presence of highly glycolytic tumor cells a metabolic competition with endothelial cells could occur, and glucose could become a limiting substrate for angiogenesis. This provisional balance between two seemingly opposing forces could be further perturbed by anti-angiogenic therapies, which hit the microvasculature and simultaneously increase tumor glycolysis. These mechanisms could underscore modulation of the therapeutic activity of anti-angiogenic drugs by a tumor metabolic trait.

Research achievements

Our team pioneered investigation of metabolic effects of anti-angiogenic drugs in solid tumors. We initially established that VEGF blockade is accompanied by dramatic reduction in glucose and ATP levels in the tumor microenvironment (Nardo G. et al. Cancer Res 2011) and this metabolic change activates the LKB1/AMPK pathway, a sensor of nutrients starvation in cells. We further hypothesized that alterations affecting this pathway could modulate therapeutic response to anti-VEGF drugs, and validated this hypothesis in CRC and lung cancer patients treated with chemotherapy plus bevacizumab (Zulato E. et al., BJC 2014 and Bonanno L. et al., CCR 2017). In parallel, we found that anti-VEGF therapy impacts on the metabolic profile of tumors and exacerbates the Warburg phenotype of tumors. Importantly, some of these metabolic changes are stable and are associated with resistance to bevacizumab (Curtarello M. et al. Cancer Res 2015). Additional metabolic changes occurring in tumors treated with angiogenesis inhibitors are currently under investigation in collaboration with other groups (Curtarello M. et al., Cells 2019).

Conclusions and perspectives

Results of our ongoing studies will provide a multi-level representation of the connections between the glycolytic phenotype of tumors and certain genetic or epigenetic profiles, highlight the therapeutic potential of glycolysis inhibitors in combination with anti-angiogenic drugs, investigate effects of tumor glycolysis on angiogenesis, and establish the possible predictive value of metabolic markers in patients treated with anti-angiogenic drugs.

NOTCH AS A THERAPEUTIC TARGET IN CANCER

The focus of this Project is to investigate the role of NOTCH signaling in cancer cells, including both T cell acute lymphoblastic leukemia (T-ALL) and certain solid tumors. Pre-clinical models are exploited to investigate the therapeutic activity of novel targeted drugs with the final aim to improve therapeutic outcome of patients with NOTCH-addicted tumors.

Background

The NOTCH pathway plays a crucial role in T-cell lineage specification and thymic development and its deregulated activation has been linked to T-ALL development as well as maintenance of key cancer features of certain solid tumors. Notably, about 50-60% of T-ALL samples show activating mutations in NOTCH1 gene and 15% of T-ALL cases present mutations or deletions in its ubiquitin ligase FBW7. The established role of NOTCH in solid and hematological malignancies suggests that this pathway could be targeted for therapeutic purposes in NOTCH-driven malignancies. Consolidated therapeutic approaches include the use of gamma secretase inhibitors (GSI) that block NOTCH processing, alternative molecules that affect NOTCH signaling and antibodies or decoy peptides to target specific NOTCH receptors or their ligands, hypothetically overcoming adverse effects due to the pan-Notch signaling inhibition associated with GSI. However, although the target has been long identifed, existing drugs have shown severe limitations in early clinical trials, including lack of efficacy and toxicity, which have so far precluded their further clinical development. There is therefore a medical need to develop new therapeutic approaches and test them in pre-clinical models to target NOTCH signaling in cancer.

Research achievements

In past years, our team pioneered in collaboration with the pediatric onco-hematology unit the set-up of patient-derived xenografts from T acute lymphoblastic leukemia (T-ALL). We utilized these well-characterized models to investigate the biological effects and the therapeutic activity of a NOTCH1 neutralizing antibody (Agnusdei V. et al., Leukemia 2014) and subsequently to dissect molecular mechanisms of resistance to NOTCH1-targeted therapy (Agnusdei V. et al., Haematologica 2019). We also observed that histone deacetylase 6 (HDAC6) controls Notch3 trafficking and degradation in T-cell acute lymphoblastic leukemia cells, uncovering that HDAC6 inhibitors can be used to counteract growth of NOTCH3-addicted tumors (Pinazza M. et al., Oncogene 2018).

Perspectives

We have patented our observation that HDAC6 inhibitors down-regulate NOTCH3 expression and activity in cancer cells and are planning to investigate the therapeutic activity of novel HDAC6 inhibitors in pre-clinical models of T-ALL as well as breast and ovarian cancer bearing NOTCH3 over-expression.

Selected publications (2006-2020):

1. Indraccolo, S., Stievano, L., Minuzzo, S., Tosello, V., Esposito, G., Piovan, E., Zamarchi, R., Chieco-Bianchi, L., Amadori, A. Interruption of tumor dormancy by a transient angiogenic burst within the tumor microenvironment (2006) Proceedings of the National Academy of Sciences of the United States of America, 103 (11), 4216-4221.

2. Indraccolo, S., Pfeffer, U., Minuzzo, S., Esposito, G., Roni, V., Mandruzzato, S., Ferrari, N., Anfosso, L., Dell'Eva, R., Noonan, D.M., Chieco-Bianchi, L., Albini, A., Amadori, A. Identification of genes selectively regulated by IFNs in endothelial cells (2007) Journal of Immunology, 178 (2), 1122-1135.

3. Moserle, L., Indraccolo, S.^§, Ghisi, M., Frasson, C., Fortunato, E., Canevari, S., Miotti, S., Tosello, V., Zamarchi, R., Corradin, A., Minuzzo, S., Rossi, E., Basso, G., Amadori, A. The side population of ovarian cancer cells is a primary target of IFN-α antitumor effects (2008) Cancer Research, 68 (14), 5658-5668.

4. Indraccolo, S., Minuzzo, S., Masiero, M., Pusceddu, I., Persano, L., Moserle, L., Reboldi, A., Favaro, E., Mecarozzi, M., Di Mario, G., Screpanti, I., Ponzoni, M., Doglioni, C., Amadori, A. Cross-talk between tumor and endothelial cells involving the Notch3-DII4 interaction marks escape from tumor dormancy (2009) Cancer Research, 69 (4), 1314-1323.

5. Nardo, G., Favaro, E., Curtarello, M., Moserle, L., Zulato, E., Persano, L., Rossi, E., Esposito, G., Crescenzi, M., Casanovas, O., Sattler, U., Mueller-Klieser, W., Biesalski, B., Thews, O., Canese, R., Iorio, E., Zanovello, P., Amadori, A., Indraccolo, S. Glycolytic phenotype and AMP kinase modify the pathologic response of tumor xenografts to VEGF neutralization (2011) Cancer Research, 71 (12), 4214-4225.

6. Agnusdei, V., Minuzzo, S., Frasson, C., Grassi, A., Axelrod, F., Satyal, S., Gurney, A., Hoey, T., Seganfreddo, E., Basso, G., Valtorta, S., Moresco, R.M., Amadori, A., Indraccolo, S. Therapeutic antibody targeting of Notch1 in T-acute lymphoblastic leukemia xenografts (2014) Leukemia, 28 (2), 278-288.

7. Curtarello, M., Zulato, E., Nardo, G., Valtorta, S., Guzzo, G., Rossi, E., Esposito, G., Msaki, A., Pastò, A., Rasola, A., Persano, L., Ciccarese, F., Bertorelle, R., Todde, S., Plebani, M., Schroer, H., Walenta, S., Mueller-Klieser, W., Amadori, A., Moresco, R.M., Indraccolo, S. VEGF-targeted therapy stably modulates the glycolytic phenotype of tumor cells (2015) Cancer Research, 75 (1), 120-133.

8. Bonanno, L., Paoli, A.D., Zulato, E., Esposito, G., Calabrese, F., Favaretto, A., Santo, A., Conte, A.D., Chilosi, M., Oniga, F., Sozzi, G., Moro, M., Ciccarese, F., Nardo, G., Bertorelle, R., Candiotto, C., Salvo, G.L.D., Amadori, A., Conte, P., Indraccolo, S. LKB1 Expression Correlates with Increased Survival in Patients with Advanced Non–Small Cell Lung Cancer Treated with Chemotherapy and Bevacizumab (2017) Clinical Cancer Research, 23 (13), 3316-3324.

9. Indraccolo, S., Lombardi, G., Fassan, M., Pasqualini, L., Giunco, S., Marcato, R., Gasparini, A., Candiotto, C., Nalio, S., Fiduccia, P., Fanelli, G.N., Pambuku, A., Puppa, A.D., D'Avella, D., Bonaldi, L., Gardiman, M.P., Bertorelle, R., De Rossi, A., Zagonel, V. Genetic, epigenetic, and immunologic profiling of MMR-deficient relapsed glioblastoma (2019) Clinical Cancer Research, 25 (6), 1828-1837.

10. Indraccolo S, De Salvo GL, Verza M, Caccese M, Esposito G, Piga I, Del Bianco P, Pizzi M, Gardiman MP, Eoli M, Rudà R, Brandes AA, Ibrahim T, Rizzato S, Lolli I, Zagonel V, Lombardi G. Phosphorylated Acetyl-CoA Carboxylase Is Associated with Clinical Benefit with Regorafenib in Relapsed Glioblastoma: REGOMA Trial Biomarker Analysis. (2020) Clin Cancer Res, Sep 1;26(17):4478-4484.

Principal Investigator (PI) of the following competitive projects (>2 MEuro since 2008):

• Ministry of Health, Ricerca Finalizzata "Angiogenesis at the interface between tumor and microenvironment: biological studies and therapeutic implications in haematologic malignancies" - RF-IOV-2006-408212, from 01-01-2008 to 31-12-2010.
• CARIPARO Foundation Excellence Project "CELLULAR INTERACTIONS IN THE BONE-MARROW VASCULAR NICHE: THE DEVELOPMENT OF AN IN VIVO MODEL TO STUDY KEY INTERACTIONS IN ACUTE LEUKEMIA", from  01-11-2009 to 01-11-2011.
• University of Padova - Progetto di Ateneo bando 2010 - CPDA103878 – “Studio della via di AMPK nella risposta dei tumori alla terapia anti-angiogenica“ from 01-01-2011 to 31-12-2012.
• AIRC - IG14295 - "Metabolic effects of anti-angiogenic therapy in cancer and their therapeutic implications " from 01-01-2014 to 31-12-2016.
• AIRC - IG18803 - "Investigating metabolic heterogeneity in tumors and its modulation by anti-angiogenic drugs" from 01-01-2017 to 31-12-2019.
• IOV Intramural Grant - bando 5x1000 anno 2018 – “Monitoring Advanced NSCLC through plasma Genotyping during Immunotherapy: Clinical feasibility and application (MAGIC-2)” from 01-01-2019 to 12-12-2021.
• AIRC - IG2020 n. 25179 - "Learning from failure: decoding metabolic traits of cancer to empower therapeutic activity of anti-angiogenic drugs" from 01-01-2021 to 31-12-2025.

PI of the following sponsored contracts (>500.000 Euro since 2012):

• Hoffman La Roche Ltd., Svizzera – “LKB1/AMPK status as biomarker of response of lung cancer to bevacizumab”, 2012.
• Oncomed Pharmaceuticals Inc., USA – “Study of the therapeutic effects of the anti-NOTCH1 antibody OMP-52M51 plus chemotherapy in preclinical models of T-ALL”, 2012.
• KYMAB Ltd., UK – “Accordo quadro per servizi di laboratorio”, 2016.
• ITALFARMACO S.p.A., Italia – “Studio dell'attività terapeutica di inibitori di HDAC6 in modelli di carcinoma ovarico e mammario ad elevata espressione di Notch3 o con mutazioni del gene ARID1“, 2019.