(P 195) THE STRATEGIC ROLE OF 3D CULTURE SYSTEMS AND ZEBRAFISH NEAR-PATIENTS PRECLINICAL PLATFORMS AS PREDICTIVE TOOLS OF CHEMOTHERAPY RESPONSE FOR SARCOMA

Author(s)

Alessandro De Vita, PhD (he/him/his)

Senior Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy.
Meldola, Emilia-Romagna, Italy

Co-Author(s)

GM

Giacomo Miserocchi, MSC

Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
Silvia Vanni, PhD (she/her/hers)

Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
Valentina Fausti, MD

Medical Doctor
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy.
Meldola, Emilia-Romagna, Italy
FP

Federica Pieri, MD

Medical Doctor
Pathology Unit, Morgagni-Pierantoni Hospital, Forlì, Italy.
Forlì, Emilia-Romagna, Italy
RC

Roberto Casadei, MD

Medical Doctor
Orthopedic Unit, Morgagni-Pierantoni Hospital, Forlì, Italy
Forlì, Emilia-Romagna, Italy
CL

Chiara Liverani, PhD

Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
CS

Chiara Spadazzi, PhD

Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
CC

Claudia Cocchi, MSC

Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
CC

Chiara Calabrese, MSC

Researcher
Osteoncology Unit, Bioscience Laboratory, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
NR

Nada Riva, MD

Medical Doctor
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
LG

Lorena Gurrieri, MD

Medical Doctor
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy
Meldola, Emilia-Romagna, Italy
FR

Federica Recine, MD

Medical Doctor
Medical Oncology Unit, Azienda Ospedaliera San Giovanni Addolorata, Roma, Italy
Rome, Lazio, Italy
AB

Alberto Bongiovanni, MD

Medical Doctor
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy.
Meldola, Emilia-Romagna, Italy
Toni Ibrahim, BSc, MD, PhD

BSc, MSc ,MD, PhD
Osteoncology, Bone and Soft Tissue Sarcomas and Innovative Therapies Unit, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy
Bologna, Emilia-Romagna, Italy
LF

Luca Frassineti, MD

Medical Doctor
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy.
Meldola, Emilia-Romagna, Italy
LC

Luana Calabrò, MD

Medical Doctor
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy.
Meldola, Emilia-Romagna, Italy
LM

Laura Mercatali, PhD

Researcher
Clinical and Experimental Oncology, Immunotherapy, Rare Cancers and Biological Resource Center, IRCCS Istituto Romagnolo Per Lo Studio Dei Tumori (IRST) "Dino Amadori", Meldola, Italy.
Meldola, Emilia-Romagna, Italy

Objective: Sarcoma suffer from the limited availability of predictive models which could guide physicians in the therapy selection. In this work with proofed the feasibility of implementing translational models for supporting daily clinical practice. In this regard sarcoma histotypes and treatments included in the clinical trial ISG10.01 were used as beta test.

Methods: This prospective study enrolled 3 patients affected by sarcoma. One undifferentiated pleomorphic sarcoma (UPS) one leyomisarcoma (LMS) and one pleomorphic liposarocma (PLS) were included. Patient-derived primary cultures of PLS were established and cultured in vitro within 3D models and in vivo through the xenostransplantation into zebrafish embryos (all included histosyeps, zebrafish n=180). These models were exposed to standard chemotherapy compared to tailored chemotherapy using the same drugs and schedule reported in the ISG10.01 trial: ifosfasmide (IFO), and epirubicin (EPI) versus gemcitabine (GEM) and docetaxel (DOCE) or gemcitabine (GEM) and dacarbazine (DACA). Cell viability was obtained by MTT and monitoring the tumor shrinkage in zebrafish using stereomicroscope microscopy and fluorescence labelled cells. Furthermore, in vivo drug toxicology was evaluated.

Results: The results confirmed the establishment of the sarcoma patient-derived primary cultures case series. Tridimensional culturing and in vivo engrafment was successfully achieved. Pharmacological profiling showed anthraciclynes-based chemotherapy as the most active treatments both in vitro and vivo. In particular PLS cultured within 3D scaffold showed a cell vialiblity of 6% with IFO+EPI and 18% with GEM+DOCE (Figure 1). Furthermore, xenostrasplanted embryos with LMS exhibited a tumor growth inhibition rate of 33.6% with IFO+EPI and 9.5% with GEM+DACA (Figure 3); with PLS 46.3% with IFO+EPI and 28.3% with GEM+DOCE; UPS showed 38.7% IFO+EPI and 14.8% GEM+DOCE 8Figure 4) all the results were statistically significant. Severe abnormalities including pericardic edema, abnormal embryonic development, enlarged yolk sack were observed in 12% of CTR PLS and 12% of GEM+DACA LMS (Figure 2). The superior activity seen with standard chemotherapy confirmed the results observed in ISG trial 10.01 corroboranting the predicitvity of these preclinical models.

Conclusion: The study aimed to provide a proof of concept of the utility of translational models including primary cultures, 3D cultures systems and zebrafish embryos as tools for chemotherapy selection into daily clinical practice. This is especially evident in rare tumors including sarcoma in which the heterogeneity of the diseases and the limited availability of predictive models make the development of new strategies an urgent clinical need. This work shed the light on the pivotal role of precision medicine which could open the window in the near future on new therapeutic horizons.

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