Research projects - Experimental Therapeutics Lab

Epigenetic mechanisms make a fundamental contribution to cancer initiation and progression. They underlie the intrinsic phenotypic plasticity of cancer cells, influence tumor–microenvironment interactions, and promote intra- and inter-tumoral heterogeneity, distant metastasis, and treatment resistance. We investigate epigenetic regulators and their roles in reshaping the transcriptomic and epigenetic landscapes during disease progression, aiming to elucidate underlying mechanisms and identify actionable targets for therapeutic intervention.

By integrating multiple experimental approaches with in vitro and in vivo models, we have gained relevant insights into critical molecular nodes that determine the efficacy and clinical potential of epigenetic compounds. Our work explores these agents both as monotherapies and in combination with other treatments, with the goal of developing novel therapeutic strategies to counteract cancer cell plasticity and the adaptive mechanisms that drive tumor progression and treatment resistance.

Relevant publications:

• Vázquez R, Civenni G, Kokanovic A, Shinde D, Cantergiani J, Marchetti M, Zoppi G, Ruggeri B, Liu PCC, Carbone GM, Catapano CV. Efficacy of Novel Bromodomain and Extraterminal Inhibitors in Combination with Chemotherapy for Castration-Resistant Prostate Cancer. Eur Urol Oncol. 2019 Aug 8:S2588-9311(19)30116-6. doi: 10.1016/j.euo.2019.07.013.
• Cacciatore A, Shinde D, Musumeci C, Sandrini G, Guarrera L, Albino D, Civenni G, Storelli E, Mosole S, Federici E, Fusina A, Iozzo M, Rinaldi A, Pecoraro M, Geiger R, Bolis M, Catapano CV, Carbone GM. Epigenome-wide impact of MAT2A sustains the androgen-indifferent state and confers synthetic vulnerability in ERG fusion-positive prostate cancer. Nat Commun. 2024 Aug 6;15(1):6672. doi:10.1038/s41467-024-50908-7. PMID: 39107274; PMCID: PMC11303763.
• Zoma M, Curti L, Shinde D, Albino D, Mitra A, Sgrignani J, Mapelli SN, Sandrini G, Civenni G, Merulla J, Chiorino G, Kunderfranco P, Cacciatore A, Kokanovic A, Rinaldi A, Cavalli A, Catapano CV, Carbone GM. EZH2-induced lysine K362 methylation enhances TMPRSS2-ERG oncogenic activity in prostate cancer. Nat Commun. 2021 Jul 6;12(1):4147. doi: 10.1038/s41467-021-24380-6.
• Federici E, Storelli E, Civenni G, Sandrini G, Guarrera L, Kokanovic A, Mosole S, Rinaldi A, Bolis M, Zhang Y, Vaddi K, Scherle P, Carbone GM, Ruggeri B, Catapano CV. Rewiring the transcriptome of castration-resistant prostate cancer through CDK9 inhibition. In revision. 2026.

Cancer cells with stem-like properties, commonly referred to as cancer stem cells (CSCs), are critical drivers of cancer evolution toward invasive, clinically aggressive, and therapy-resistant disease. These cells possess self-renewal capacity, tumor-initiating properties, and enhanced phenotypic plasticity. Moreover, CSCs are intrinsically resistant to many anticancer therapies, contributing to treatment failure and tumor relapse.

The development of drugs that specifically target tumor-initiating stem-like cells could represent an important advance in cancer treatment. Increasing attention is therefore being directed toward the biological pathways that enable cancer cells to adapt to and survive microenvironmental stress while maintaining their tumor-initiating and metastatic properties. A deeper understanding of these pathways may reveal critical molecular nodes and targetable vulnerabilities for the development of innovative CSC-directed therapeutic strategies.

Mitochondrial dynamics

Mitochondrial reprogramming and metabolic pathways are critical for maintaining cancer cell phenotypic plasticity and survival. Mitochondrial homeostasis depends on a finely regulated balance among mitochondrial dynamics, biogenesis, and clearance.

We have shown that mitochondrial dynamics, characterized by cycles of fission and fusion, are essential for the appropriate partitioning of healthy mitochondria between daughter stem cells and for preserving self-renewal capacity. Proteins involved in the mitochondrial dynamics machinery, including Drp1 and MFF, therefore represent promising targets for the development of novel CSC-directed therapies.

However, the mechanisms responsible for enhanced mitochondrial dynamics in prostate cancer, as well as the functional consequences of disrupting these processes, remain incompletely understood. Our work suggests that mitochondrial dynamics and metabolism generate signals that can either promote or suppress the survival and proliferative capacity of cancer stem-like cells.

A major objective of our work is to define the relationships among mitochondrial plasticity and dynamics, cellular metabolism, and CSC-sustaining pathways and to identify their specific molecular effectors. These studies aim to address important knowledge gaps regarding the role of mitochondria in cancer and to provide a foundation for developing innovative CSC-directed therapies.

Mitochondrial inter-organelle communication

Our current research also investigates how mitochondria communicate with other organelles, including the nucleus, endoplasmic reticulum (ER), and lipid droplets, to support cancer cell survival. We aim to elucidate the relationships among mitochondrial dynamics, metabolic pathways, and epigenetic mechanisms. Factors that regulate inter-organelle communication and mitochondrial homeostasis may provide additional targets for drug discovery.

The σ1 receptor is a ligand-activated molecular chaperone preferentially localized at the ER and at mitochondria-associated ER membrane (MAM) domains. We have shown that the σ1 receptor plays an essential role in castration-resistant prostate cancer through its involvement in ER–mitochondrial communication, stress responses, metabolic reprogramming, and mitochondrial dynamics.

We are extending these findings by investigating the effects of genetic knockdown and small-molecule inhibitors of the σ1 receptor on tumor-initiating stem-like cells across multiple cancer types.

Relevant publications: 

• Pandit SK, Sandrini G, Merulla J, Nobili V, Wang X, Zangari A, Rinaldi A, Shinde D, Carbone GM, Catapano CV. Mitochondrial Plasticity Promotes Resistance to Sorafenib and Vulnerability to STAT3 Inhibition in Human Hepatocellular Carcinoma. Cancers 2021 Nov 30;13(23):6029. doi: 10.3390/cancers13236029. PMID: 34885140; PMCID: PMC8657239.
• Civenni G, Albino D, Shinde D, Vázquez R, Merulla J, Kokanovic A, Mapelli SN, Carbone GM, Catapano CV. Transcriptional Reprogramming and Novel Therapeutic Approaches for Targeting Prostate Cancer Stem Cells. Front Oncol. 2019 May 9;9:385. doi: 10.3389/fonc.2019.00385.
• Civenni G, Bosotti R, Timpanaro A, Vàzquez R, Merulla J, Pandit S, Rossi S, Albino D, Allegrini S, Mitra A, Mapelli SN, Vierling L, Giurdanella M, Marchetti M, Paganoni A, Rinaldi A, Losa M, Mira-Catò E, D'Antuono R, Morone D, Rezai K, D'Ambrosio G, Ouafik L, Mackenzie S, Riveiro ME, Cvitkovic E, Carbone GM, Catapano CV. Epigenetic Control of Mitochondrial Fission Enables Self-Renewal of Stem-like Tumor Cells in Human Prostate Cancer. Cell Metab. 2019 Aug 6;30(2):303-318.e6. doi: 10.1016/j.cmet.2019.05.004.
• Kopp N, Holtschulte C, Börgel F, Lehmkuhl K, Friedland K, Civenni G, Laurini E, Catapano CV, Pricl S, Humpf HU, Schepmann D, Wünsch B. Novel σ1 antagonists designed for tumor therapy: Structure-activity relationships of aminoethyl substituted cyclohexanes. Eur J Med Chem. 2021 Jan 15;210:112950. doi: 10.1016/j.ejmech.2020.112950.
• Civenni G, Sandrini G, Merulla J, Musumeci C, Federici E, Vallegra A, Kokanovic A, Mosole S, Shinde D, Sorrenti E, Paganoni AJJ, Marchetti M, Valzelli R, Albino D, Pecoraro M, Rinaldi A, Bolis M, Geiger R, Winge T, Holtschulte C, Laurini E, Pricl S, Carbone GM, Wünsch B, Catapano CV. Integrated control of cancer stemness by σ1 receptor in advanced prostate cancer. Oncogene. 2025 Nov;44(42):4032-4046. doi: 10.1038/s41388-025-03541-7. Epub 2025 Sep 2. PMID: 40897798; PMCID: PMC12518131.

Liver cancer is the second leading cause of cancer-related death, and its incidence is increasing worldwide. Hepatocellular carcinoma accounts for approximately 90% of liver cancer cases, and therapeutic options for advanced disease remain limited. The altered metabolic and immune microenvironment associated with liver cancer plays an important role in disease progression and treatment failure.

Lin28A/B are RNA-binding proteins that repress the biogenesis of let-7 miRNAs or enhance the translation of specific mRNAs. Lin28 proteins are upregulated in many cancers, where they promote malignant transformation by increasing the expression of multiple oncogenes and stem cell factors and thereby expanding poorly differentiated, stem-like tumor cell populations.

The Lin28/let-7 axis has therefore attracted considerable interest as a potential therapeutic target across multiple cancer types. Recent work has also positioned Lin28 at the interface between tumorigenesis and metabolic reprogramming.

Our current findings suggest that selective interference with Lin28A/B proteins may have beneficial effects in liver metabolic diseases and cancer. We found that Lin28 inhibition represses lipogenesis and stimulates ketogenesis in the mouse liver, resulting in a sustained increase in ketone levels, including β-hydroxybutyrate.

We are currently investigating the impact of Lin28 targeting on metabolic reprogramming using human organoids and xenografts, as well as murine organoids, allografts, and genetically engineered mouse models of hepatocellular carcinoma. Through these studies, the project aims to support the development of novel and potent therapeutic approaches for advanced hepatocellular carcinoma and other cancers.

Relevant publications:

• Albino D, Civenni G, Dallavalle C, Roos M, Jahns H, Curti L, Rossi S, Pinton S, D'Ambrosio G, Sessa F, Hall J, Catapano CV, Carbone GM. Activation of the Lin28/let-7 Axis by Loss of ESE3/EHF Promotes a Tumorigenic and Stem-like Phenotype in Prostate Cancer. Cancer Res. 2016 Jun 15;76(12):3629-43. doi:10.1158/0008-5472.CAN-15-2665. Epub 2016 May 2. PMID: 27197175.
• Roos M, Rebhan MA, Lucic M, Pavlicek D, Pradere U, Towbin H, Civenni G, Catapano CV, Hall J. Short loop-targeting oligoribonucleotides antagonize Lin28 and enable pre-let-7 processing and suppression of cell growth in let-7-deficient cancer cells. Nucleic Acids Res. 2015 Jan;43(2):e9. doi:10.1093/nar/gku1090. Epub 2014 Nov 6. PMID: 25378324; PMCID: PMC4333367.
• Lekka E, Kokanovic A, Mosole S, Civenni G, Schmidli S, Laski A, Ghidini A, Iyer P, Berk C, Behera A, Catapano CV, Hall J. Pharmacological inhibition of Lin28 promotes ketogenesis and restores lipid homeostasis in models of non-alcoholic fatty liver disease. Nat Commun. 2022 Dec 26;13(1):7940. doi:10.1038/s41467-022-35481-1. PMID: 36572670; PMCID: PMC9792516.
• Laski A, Brümmer A, Lucic M, Federici E, Mestres-Pascual I, Imig J, Kanitz A, Zavolan M, Gebert LFR, MacRae IJ, Catapano CV, Hall J. Seedless 3' pairing enables miR-17 family miRNAs to seize let-7 target sites. Nucleic Acids Res. 2026 Aug 10;54(15):gkag767. doi: 10.1093/nar/gkag767.PMID: 42598859.

Despite an initial response to androgen receptor-directed therapies, many patients with prostate cancer ultimately progress to the hormone-refractory stage, for which current treatment options remain inadequate.

Antibody-drug conjugates (ADCs) directed against tumor-associated cell-surface proteins represent a promising precision-medicine strategy. By enabling selective drug delivery to tumor cells, ADCs have the potential to enhance therapeutic efficacy while minimizing off-target effects.

We have investigated the expression of several targetable cell-surface proteins in clinical samples, human cancer cell lines, patient-derived organoids, and xenograft models of prostate cancer, assessing inter- and intra-tumoral heterogeneity and co-expression patterns across these models and samples.

In parallel, we are evaluating engineered ADCs targeting specific cell-surface receptors, including PSMA. These ADCs undergo rapid internalization, efficiently release their cytotoxic payloads, and display selective activity against receptor-positive cancer cells while sparing receptor-negative cells in 2D cultures, 3D tumor organoids, and in vivo models. Notably, administration of these ADCs to tumor-bearing mice induces durable tumor regression in xenograft models of hormone-refractory prostate cancer with minimal toxicity. These findings provide valuable information for selecting appropriate targets for antibody-based therapies and a strong rationale for developing targeted strategies for advanced prostate cancer.

In related collaborative projects, we are also applying innovative antibody-based approaches to engineer nanoparticles for therapeutic payload delivery and to address poorly druggable anticancer targets. By combining the potency and selectivity of antibodies with recent advances in nanodelivery systems, we aim to develop a new generation of therapeutic payload-delivery strategies that address the biological complexity and phenotypic diversity of human cancers.

Relevant publications:

• Daniela Impellizzieri, Elisa Storelli, Atik Balla, Simone Mosole, Cristina Dongilli, Federico Jauk, Roberta Frapolli, Lavinia Morosi, Maurizio D'Incalci, Jemila Houacine, Morris Rosenberg, Mohamed Bekradda, Esteban Cvitkovic, Carlo V. Catapano. Long-term tumor growth inhibition and extended survival with TD001, a novel optimized PSMA-targeting ADC, in PSMA-expressing CRPC CDX castrated mouse models. AACR annual meeting, Chicago, USA, 2025
• Daniela Impellizzieri, Elisa Storelli, Atik Balla, Simone Mosole, Cristina Dongilli, Federico Jauk, Roberta Frapolli, Lavinia Morosi, Maurizio D'Incalci, Jemila Houacine, Morris Rosenberg, Bérangère Deleglise, Esteban Cvitkovic, Carlo V. Catapano. Improved tumor penetration and cytotoxic payload release with TD001, a novel PSMA-targeting ADC with optimized linker-payload composition, in PSMA-expressing CRPC CDX castrated mouse models. AACR annual meeting, Chicago, USA, 2025
• Domenico Albino, Carola Musumeci, Elisa Storelli, Atik Balla, Elisa Federici, Gianluca Civenni, Daniela Impellizzieri, Giada Andrea Cassanmagnago, Marco Bolis, Steve Pascolo, Carlo V Catapano, Giuseppina MR Carbone. Therapeutic Reversal of Tumor Cell Plasticity by EHF mRNA Delivery in Prostate Cancer. AACR Drug Discovery and Development (AACR D3), July 21 - 24, 2026, Boston, Massachusetts, USA.
• Carola Musumeci, Concetta Guerra, Domenico Albino, Elisa Storelli, Jacopo Sgrignani, Daniela Impellizieri, Simone Moro, Elisa Federici, Andrea Cavalli, Giuseppina MR Carbone, Carlo V Catapano. Suppressing ERG-driven oncogenesis by mRNA-based delivery of an intracellular minibody targeting a unique protein methylation site in ERG fusion-positive prostate cancer. AACR Drug Discovery and Development (AACR D3), July 21 - 24, 2026, Boston, Massachusetts, USA.

Related collaborative project: R&D AbITi

New therapeutic options are needed for patients with metastatic cancers that no longer respond to current treatments. Intratumoral heterogeneity and epigenetic variability among tumor cell subpopulations promote the emergence of treatment-resistant clones.

Preclinical evidence indicates that combining drugs with distinct mechanisms of action, such as epigenetic agents and conventional chemotherapeutics, may improve therapeutic outcomes in metastatic cancer. Although concomitant administration of these drug combinations can result in strong synergistic effects, cumulative toxicity may limit their clinical applicability.

We aim to develop nanoparticle-based strategies that combine synergistic therapeutic payloads to selectively and effectively target metastatic cancer. This nanomedicine approach, based on multi-drug-loaded nanoparticles, may enhance therapeutic efficacy by enabling the simultaneous and targeted delivery of rationally designed drug combinations to cancer cells while reducing systemic exposure and toxicity in normal tissues.

Metastatic dissemination to distant organs is a major cause of cancer-related mortality. Prostate cancer is the second most common cancer and a leading cause of cancer-related death among men. Although localized prostate tumors are rarely lethal, prognosis worsens dramatically once the disease progresses and metastasizes to distant organs, including bone, lymph nodes, liver, and lung.

Although liver and lung metastases are uncommon at initial diagnosis, their frequency increases in heavily treated patients, and their occurrence is associated with poor prognosis and marked resistance to therapy. Despite their clinical relevance, limited information is available regarding the intrinsic phenotypic characteristics of tumor cells and the extrinsic, organ-specific factors that promote metastatic colonization of these sites. Reliable experimental models of prostate cancer visceral metastasis and specific therapeutic strategies for this setting are also lacking.

We have established models, including 3D tumor organoids and assembloids, human xenografts, and murine allografts, to investigate prostate cancer metastasis to visceral organs such as the liver. Using these systems, we are studying the mechanisms underlying metastatic colonization and progression.

Our studies aim to identify the relevant biological processes and specific molecular targets that can be addressed with novel therapeutic approaches. In this context, we are investigating several strategies, including small-molecule drugs, RNA/DNA therapeutics, and antibody-based therapies.

Related collaborative project: NERD