Patient-derived Tumor Xenograft Models Research Articles

Abstract 3039: Essential role of IRAK4/TPL2 signaling axis in MAPK activation by oncogenic RAS and genotoxic stress

Abstract Pancreatic ductal adenocarcinoma (PDAC) has a 5-year survival of 9% and effective treatment options remain elusive. Oncogenic mutations of KRAS occur in >95% of PDACs and are well-established as the bona fide driver event. However, inhibition of KRAS oncoprotein or its downstream signaling cascades remains unsuccessful in PDAC patients. Furthermore, parallel survival pathways including constitutive activation of the NF-κB pathway poses an additional therapeutic barrier. Previous work from our lab showed that Interleukin-1 Receptor associated kinase 4 (IRAK4) is a major driver of NF-kB cascade in PDAC. Here, through an unbiased reverse phase protein array screen and RNA sequencing, we discovered IRAK4 controls MAPK activity downstream of KRAS. Ablation of IRAK4 completely abolishes RAS-induced transformation in human and murine cell lines. Mechanistically, we implicate a KRAS-driven IL-1β signaling loop that activates IRAK4 and uncover MAP3K8 (or TPL2/COT) as the kinase through which IRAK4 activates MEK and ERK. Suppression of TPL2 abrogates KRAS-driven MEK-ERK activity and transformed growth of PDAC cell lines. In addition, TPL2 inhibition suppresses p105/p50 NF-kB activation, a valuable phenomenon that distinguishes TPL2 inhibition from MEK inhibition. We find TPL2 inhibition synergizes with chemotherapy to suppress growth of PDAC cell lines in vitro and patient-derived xenograft tumor model in vivo. Analyses of PDAC tissue microarray showed TPL2 expression to be marginally associated with poor prognosis. Additionally, we are the first to characterize gain-of-function point mutations in TPL2 which hyperactivate MAPK and NF-kB, in part by preventing TPL2 protein degradation. Together, our study broadens the understanding of the oncogenic RAS signaling network and reveals IRAK4 and TPL2 as novel practical therapeutic targets in RAS-driven cancers. Citation Format: Paarth B. Dodhiawala, Namrata Khurana, Daoxiang Zhang, Yi Cheng, Lin Li, Kuljeet Seehra, Hongmei Jiang, Patrick M. Grierson, Andrea Wang-Gillam, Marianna B. Ruzinova, Kian-Huat Lim. Essential role of IRAK4/TPL2 signaling axis in MAPK activation by oncogenic RAS and genotoxic stress [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3039.

Abstract 5046: Natural Killer (NK) human immune system (HIS) immunograft platform to evaluate the pharmacodynamics of immuno-oncology therapeutics

Abstract Background: Harnessing NK cell anti-cancer cytotoxicity has gained interest as a therapeutic strategy, and consequently improved preclinical models supporting the translation of NK cell-mediated therapies to the clinic are desired. Reproducible models with human NK engraftment into immunodeficient mice co-engrafted with cell line-derived xenograft or patient-derived xenograft tumor models have been lacking due to an inability to support NK cell engraftment and persistence. Here we evaluated IL-15-NOG mice for the engraftment and sustained survival of both ex vivo expanded and primary human NK cell isolates for establishing models that engraft effectively with both human NK cells and a PDX or CDX tumor. Materials and Methods: NK cells from normal adult peripheral blood mononuclear cells (PBMC) donors (N=3) were expanded using two different commercially available kits and evaluated for NK phenotype, expansion rates and yields. Ex vivo expanded NK cells were tested for antibody dependent cell cytotoxicity (ADCC). Titrated doses of ex vivo expanded NK cells were adoptively transferred into IL-15-NOG mice for human chimerism, and the persistence and survival of NK cells and their immunophenotype were assessed. In separate studies, naïve NK cells enriched from PBMC were also evaluated for NK cell persistence and expansion in vivo. To establish an NK ImmunoGraft, NK cells were engrafted in xenograft tumor bearing mice (CDX or PDX) and tumor growth kinetics and infiltrating NK cells phenotype were characterized. Results: Donor dependent NK expansion was observed ex vivo, with 28 to 50-fold expansion by two weeks. NK cells expanded ex vivo were CD3−CD16+CD56± and varied based on the expansion kit utilized. ADCC mediated lysis was observed with ex vivo expanded NK cells. Nearly all CD45+ cells in circulation were NK cells, and these peaked by week 2, and were maintained for up to 10 weeks in IL-15-NOG mice. Primary NK cells engrafted with slower kinetics, with peak abundance at 3-4 weeks. NK cells expressed granzyme B, and further functional studies are in progress. For all NK cell populations, cell density-dependent engraftment was observed with a largely stable NK phenotype observed across the study. In the absence of any therapeutic treatment, NK cell persistence and expansion in vivo did not inhibit tumor xenograft growth kinetics in IL-15-NOG mice. Conclusions: IL-15-NOG mice support the survival and persistence of human NK cells from both ex vivo expanded and naïve NK cells, suggesting the universality of this platform for human NK engraftment. Our preliminary studies support IL-15 NOG mouse model as a suitable system for evaluation of NK cellular therapies or NK cell-modulating therapies in the context of patient-derived or cell-line derived xenograft (PDX or CDX) mouse models. Citation Format: Bhavna Verma, Jon Weidanz, Amy Wesa. Natural Killer (NK) human immune system (HIS) immunograft platform to evaluate the pharmacodynamics of immuno-oncology therapeutics [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5046.

Abstract 6124: Translation of zebrafish tumor-derived xenograft-models for improved diagnosis and treatment planning in urinary bladder cancer patients

Abstract Precision medicine in oncology aims to identify the most effective treatment for any given patient based on individualized analyses of patient material. Currently, precision medicine relies on sequencing of DNA or RNA to identify patient tumor-specific mutational profiles that may be coupled to drug response. These techniques, however, fail to reveal actionable mutations in approximately 85% of the cancer patients, and have not been established at all for many commonly used drugs including cisplatin-based treatments used in urinary bladder cancer. While mouse-PDX models can determine drug response rates with high accuracy in most patients and for most drugs, such techniques are too slow and expensive to be relevant for first line treatment planning. Urinary bladder cancer patients are often treated with cisplatin-containing combination therapy, with the hope of down-staging tumors before surgery. 60%, however, do not respond or even progress on this treatment, and these patients would benefit from immediate surgery upon diagnosis. To help identify non-responding patients, we show here that patient-derived tumor xenograft models can be established in zebrafish larvae (ZTX models) and that the resulting tumors exhibit differential responses to the two main cisplatin-containing treatments GC and MVAC. Preliminary results from the first 19 patients are presented. Two tumor biopsies were destroyed during transport and two did not allow isolation of sufficient viable cells for implantation. From the remaining 15 samples an average of 2,6 million cells with average viability of 53% were isolated and used to implant at least 60 2-days old larvae. All 15 samples implanted in the larvae and survived and/or grew exhibiting varying degrees of metastatic dissemination (average between 2 and 13 metastasized cells per embryo and model) within only three days from implantation. Four ZTX models exhibited different responses to GC and MVAC demonstrating that these treatments are not equally effective in all patients. Non-response in ZTX models was associated with tumors having re-appeared in the bladder upon radical cystectomy in all patients undergoing surgery prior to Dec. 5th 2019 (n=3). GC inhibited metastasis in all models (average 69% inhibition), whereas MVAC inhibited metastasis in 40% of the models (average 36% inhibition). In conclusion: The ZTX urinary bladder cancer platform presented here overcome limitations associated with long assay time and high cost of other functional models within precision medicine as well as the low hit-rate of actionable mutations associated with genomic techniques. ZTX models will therefore likely become a powerful method for functional precision medicine within oncology, in the near future. Citation Format: Zaheer Ali, Anna Nilsson, Malin Vildevall, Larissa Rizzo, Ylva Huge, Amir Sherif, Anna Fahlgren, Lasse DE Jensen. Translation of zebrafish tumor-derived xenograft-models for improved diagnosis and treatment planning in urinary bladder cancer patients [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6124.

Abstract 1672: Comparing subcutaneous to mammary fat pad implantation on the growth kinetics, histology and drug response of PDX breast cancer models

Abstract Preclinical oncology studies frequently use patient-derived xenograft (PDX) tumor models and the efficacy of the test therapy is evaluated by measuring changes in tumor volume. Subcutaneous (SQ) implantation of tumors is most widely used in oncology studies due to ease of technique and tumor accessibility for caliper measurements. However, SQ tumors are then growing in a different environment compared to the tumor model. Conversely, orthotopic implantation provide tumors with a similar microenvironment for growth, but require surgical techniques which increase the amount of resources, time, and skill needed to perform these studies. Currently, the standard method for an orthotopic breast cancer involves surgically clearing the mammary fat pad (MFP) prior to engrafting the cells or tissue. In this study we developed a nonsurgical approach where PDX tissue is engrafted directly into a non-cleared MFP using a method similar to SQ engraftment. Using several different established breast cancer PDX models, this study compares the impact of SQ and MFP implantation on growth kinetics, tumor histology and drug efficacy. For each experiment, 8-week old female NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG; JAX strain #005557) mice were shaved on the right flank or MFP, earnotched and housed. PDX breast tumor tissue from donor mice was cleared of necrotic tissue and mouse tissue, minced and loaded into syringes capped with either 14G or 18G trocar needles. Mice were put under anesthesia and trocared with 50ul of PDX breast tumor model tissue. Non-cleared implantations were performed in all groups and tumor models. Tumor growth was monitored using caliper measurements until the study terminus. Slides generated from formalin-fixed and paraffin embedded tumor tissue were used for histological analysis. Cisplatin and Docetaxel were selected as standard of care agents to compare drug responses in SQ versus MFP engrafted tumors. In all tested models, tumors volumes in the MFP cohort are significantly higher compared to tumors implanted SQ. Tumors engrafted into the MFP reached 500mm3 significantly earlier than tumors engrafted SQ, but the doubling times remained unchanged. This suggests the environment of the MFP allows for a faster engraftment rather than a shorter doubling time. H&E staining shows no significant histological difference between tumor models engrafted in the SQ and MFP for this study. This likely because tumor models were established in the SQ for at least 3 rounds of engraftment before being engrafted in the MFP for this study. Histological differences may have been prominent if the tumor models were established in the appropriate locations (SQ or MFP) at model development. This study has important implications for ensuring a more clinically relevant growth environment for mouse studies using PDX breast models. Citation Format: Kyle M. Draheim, Raphaela R. Banzon, Marta M. Tewodros, Daniel VanBuskirk, Mingshan Cheng. Comparing subcutaneous to mammary fat pad implantation on the growth kinetics, histology and drug response of PDX breast cancer models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1672.

Abstract LB-110: Computational discovery and preclinical validation of therapeutic leads with novel MOAs for hepatocellular carcinoma and pancreatic ductal adenocarcinoma

Abstract Hepatocellular carcinoma (HCC) and pancreatic ductal adenocarcinoma (PDAC) have among the lowest 5-year survival rates of all cancer types at 18% and 9%, respectively. Treatment options for patients with liver or pancreatic cancer are relatively unchanged over the past 10 years. HCC has seen the recent FDA-approval of multi-kinase inhibitor therapies with similar mechanisms of action, including cabozantinib, regorafenib, and lenvatinib, and the immune checkpoint inhibitor nivolumab (conditionally). Despite these advances, the survival rate and median survival time for HCC patients remain poor. The picture for PDAC patients is similar, although with even greater need for new therapies. We present results from a powerful and efficient computational drug discovery platform that produces drug discovery hits with first-in-class mechanisms of action that can advance rapidly and successfully through preclinical validation studies. The twoXAR discovery platform uses an artificial-intelligence framework to integrate diverse patient-derived data sets and build holistic and unbiased models of human disease biology. The utilization of diverse, proprietary algorithms and deep learning principles provides a highly sensitive platform to elucidate detailed disease-specific associations between biology and biomedical data that are integrated with a library of existing drug molecules to deliver novel, high-value drug discovery hits. The twoXAR platform delivers drug discovery hits with known pharmacological properties and preserves the data-driven links to disease biology; this facilitates validation and optimization studies. We employed the twoXAR platform to build in-silico disease models of HCC and PDAC using disease-specific data and generated a set of 10 molecules with predicted efficacy in HCC and a second, independent set of 11 molecules with predicted efficacy in PDAC. These independent sets of disease-specific drug discovery hits represented novel mechanisms of action that had not been tested previously as potential clinical therapies for HCC or PDAC, respectively. TXR-311 and TXR-312, and TXR-405 and TXR-411 were discovered as validated hits for HCC and PDAC, respectively, using in vitro cell proliferation and viability assays with HCC and PDAC tumor cell lines. In these studies, TXR-311 inhibited proliferation and viability of five different HCC tumor cell lines with IC50 values that were 70-fold lower than IC50 values for sorafenib and displayed greater than 500-fold selectivity against primary human hepatocytes. In subsequent in vivo efficacy studies using two HCC patient-derived xenograft (PDX) tumor models, TXR-311 showed excellent tolerability and displayed significant tumor growth inhibition efficacy compared to vehicle-treated controls. TXR-311 presents a first-in-class lead for further development as a potential HCC therapy. Citation Format: Isaac Hakim, Mei-Sze Chua, Wei Wei, Li Ma, Elizabeth Noblin, Samuel So, Aaron C. Daugherty, Timothy S. Heuer. Computational discovery and preclinical validation of therapeutic leads with novel MOAs for hepatocellular carcinoma and pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr LB-110.

Abstract 5054: Synergistic anti-cancer activity of palbociclib in combination with sunitinib in patient-derived xenograft (PDX) models of various human cancers types

Abstract Introduction: Treatment for cancer, particularly in later lines, is often inadequate. Combinational therapies targeting several signaling pathways simultaneously are considered a promising approach, with limited clinical evidence. Effectiveness of such combinations can be explored in patient-derived xenograft (PDX) tumor models, which more accurately represent human tumor biology, drug responsiveness, and treatment-related toxicity than standard xenograft models derived from established human cancer cell lines. Objectives: To evaluate, in PDX models, the anti-tumor effect and safety of combinational therapy including a CDK4/6 inhibitor (palbociclib) plus a multi-targeted receptor tyrosine kinase inhibitor (mtRTKI; sunitinib). Methods: Overall, 20 PDX models were developed from patients with various solid tumors. For each, tumor was implanted subcutaneously in immune-deficient mice (NRG or NSG). The mice were randomized to 4 treatment groups: vehicle as control, palbociclib (100 mg/kg), sunitinib (50 mg/kg), or combination thereof. Each treatment group included 5-6 mice (18/20 models). Drugs were administered orally, 5 days per week. Tumor volume was monitored twice a week with digital caliper. Mice body weight and clinical signs were examined twice a week to evaluate toxicity. Results: The mean experiment time (all experiments) was 66 days (SEM, 10 days; range, 15-170 days). In 14 of the 20 evaluated models (70%), including those for cholangiocarcinoma, breast, lung, pancreas, colon, stomach, and ovarian cancer, combinational therapy with palbociclib and sunitinib had significantly superior anti-cancer activity compared to either drug alone, without any unexpected toxicity. Mean duration of response in these 14 models was 72 days (SEM, 12 days). The combination of palbociclib and sunitinib prevented tumor growth, and in 3 models reduced tumor size. In the 2 PDX models developed from patients with palbociclib-resistant disease, the combination overcame tumor resistance to palbociclib. In 5 of the models which demonstrated synergy between palbociclib and sunitinib, the effect of administrating 2-fold lower doses of both drugs was evaluated. The lower doses were shown to suppress tumor growth in a dose-response manner. Conclusions: These PDX results suggest that the combination of palbociclib and sunitinib exert a synergistic anti-tumor effect on multiple tumor types without adding unexpected toxicity. Citation Format: Neta Moskovits, Idit Peretz, Eva Chausky, Raisa Meerson, Ella Itzhaki, Noga Marsiano, Nofar Shmuel, Ranny Yaffe, Natalia Edison, Tal Goldman, Salomon Stemmer. Synergistic anti-cancer activity of palbociclib in combination with sunitinib in patient-derived xenograft (PDX) models of various human cancers types [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5054.

Abstract 6223: Combination of BCL-2/BCL-xL dual inhibitor APG-1252 and chemotherapeutics overcomes resistance to osimertinib in EGFR mutant NSCLC in preclinical models

Abstract Osimertinib (AZD9291) is the first-line treatment for EGFR-mutated NSCLC; however, the majority of patients inevitably develop resistance due to de novo genomic abnormalities, such as C797S mutation, EGFR exon 20 insertion, MET amplification and other unknown mechanisms. Hence, effective therapies to overcome acquired resistance are urgently needed. Inhibition of BCL-2/BCL-xL has been reported to enhance apoptosis in EGFR-TKI resistant cells with low sensitivity to EGFR inhibition. In this study, we evaluated whether the combination of a dual BCL-2/BCL-xL inhibitor APG-1252 and chemotherapeutics could overcome osimertinib resistance in preclinical xenograft models. First, EGFR C797S mutation was introduced to human NCI-H1975 NSCLC cells to construct an NCI-H1975-C797S resistant cell line using CRISPR technology. NCI-H1975-C797S derived-xenografts were treated with APG-1252, cisplatin/docetaxel or their combinations. Combination therapy with APG-1252 and cisplatin or docetaxel exhibited synergistic antitumor activity. APG-1252 plus docetaxel combination achieved 100% tumor partial regression (PR). Similar results were demonstrated in a patient-derived xenograft (PDX) tumor model derived from an osimertinib-resistant NSCLC patient harboring 19del-T790M-C797S mutations. Furthermore, the combinations also exhibited enhanced antitumor activity in an osimertinib-resistant PDX model that the resistant mechanism remained unknown. In summary, our results suggest that the combination treatment with APG-1252 and chemotherapeutics can overcome acquired resistance to osimertinib and the combination deserves further clinical evaluations. Citation Format: Ran Tao, Guangfeng Wang, Douglas D. Fang, Guoqin Zhai, Yuanbao Li, Jing Lv, Miaoyi Wu, Yangfeng Ge, Feifei Zhang, Danyi Wen, Dajun Yang, Yifan Zhai. Combination of BCL-2/BCL-xL dual inhibitor APG-1252 and chemotherapeutics overcomes resistance to osimertinib in EGFR mutant NSCLC in preclinical models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6223.

Abstract 6126: Zebrafish patient tumor-derived xenograft models used for pre-clinical evaluation of CAN04 for lung and pancreatic cancer

Abstract Lung cancer is one of the most deadly diseases accounting for 24% of all cancer deaths worldwide. One reason for this high mortality is the high interindividual heterogeneity but generally poor efficacy of current treatments, leading to an urgent need for new and more effective drugs. Understanding the individual variability in the efficacy of new treatment candidates, delineating whether they should be combined with existing chemotherapeutics and to what extent they affect metastatic dissemination of the tumor cells are key preclinical indicators needed to increase the chance of success in clinical trials. Developing such data, however, requires animal models that recapitulate individual differences of different lung cancer patients, include insights into metastatic activity and allow analysis of a large amounts of treatment combinations for each patient model. As such, an in vivo screening system which has higher throughput than mouse models and at the same time allows analysis of metastatic activity would be very valuable in mimicking human disease. Here we conducted zebrafish patient tumor derived xenograft (PDX)-studies based on cisplatin sensitive and -resistant lung cancer PDX material, to test the efficacy of a a novel antibody, CAN04, under development for this indication. CAN04 targets Interleukin-1 Accessory Protein (IL1RAP) and has shown synergistic effects with cisplatin in murine models of cancer. CAN04 is currently in phase II development in combination with chemotherapy in lung cancer and pancreatic cancer. CAN04 was given either alone or with cisplatin at three different concentrations, and the effects on primary tumor growth and metastasis three days after tumor implantation was evaluated. We show that CAN04 was able to synergize with cisplatin in causing almost complete (85%-98%) tumor regression even of cisplatin-resistant tumors, compared to non-treated controls. The effects were concentration- and model-dependent. Interestingly, in the cisplatin-resistant model, the antibody and cisplatin co-treatment led to robust inhibition of metastatic dissemination, which was not seen in either group alone. This substantiates the beneficial therapeutic efficacy of combining CAN04 to cisplatin treatment in lung cancer. In conclusion, zebrafish-PDX (ZTX) models are powerful tools for evaluating individual differences in drug sensitivity on both primary tumor growth and metastasis and are suitable for screening various drug concentrations and/or combinations in multiple models with results being generated within one or a few weeks. We further conclude that CAN04 is inducing cisplatin sensitivity and synergize with cisplatin to inhibit metastasis, at least in some cisplatin resistant lung cancers. Citation Format: Zaheer Ali, Anna Nilsson, Malin Vildevall, Julia Schueler, David Liberg, Anna Fahlgren, Lasse DE Jensen. Zebrafish patient tumor-derived xenograft models used for pre-clinical evaluation of CAN04 for lung and pancreatic cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6126.

Abstract 6125: Zebrafish patient tumor-derived xenograft models synergize with mouse-PDX models for understanding variation in anti-cancer drug responses

Abstract Lung cancer accounts for the 2nd most common cancer among men and women, representing 24% of cancer deaths worldwide. Standard-of-care treatments vary considerably depending on the tumor type and staging. Identifying which patients will benefit from treatment with a certain drug remains one of the major challenges in the clinic. Genetic analyses are widely used but have low applicability as only ~10% of the patients have mutations coupled to available targeted therapies and relatively low sensitivity as therapeutic effects are absent in ~50% of the predicted responders. Mouse-PDX models can accurately determine drug response rates for 50-60% of the patients, but are not well suited for evaluating metastatic risk. As metastasis is a major cause of disease-associated mortality and no drugs that target metastasis exist today, there is considerable need to develop new drugs able to impair metastatic dissemination in lung (and other) cancers. To meet this need, zebrafish-PDX (ZTX) models are ideally positioned as a synergistic complement to mouse-PDX models allowing evaluation of drug responses, in a non-rodent in vivo system with the turnaround time and scalability of an in vitro platform Here we generated zebrafish- and mouse-PDX models based on 20 patient NSCLC samples and compared the efficacy of standard-of-care treatment (erlotinib and paclitaxel) on primary tumor growth/regression as well as metastatic dissemination in the zebrafish-PDX models. The ZTX models exhibited variable sensitivity to the drugs tested with 11 of 20 and 16 of 20 models being sensitive to erlotinib and paclitaxel respectively. The efficacy of erlotinib and/or paclitaxel was identical in 9 of 11 mouse- and zebrafish-PDX models where these drugs were compared head-to-head. The models metastasized within three days of tumor implantation in the zebrafish larvae, seeding an average of 2 - 8 metastatic lesions per model in the caudal hematopoietic plexus. Paclitaxel and erlotinib inhibited metastasis in 7 of 20 and 6 of 20 models respectively. The anti-metastatic activity did not correlate with the activity against the primary tumor. Investigations as to what extent this correlates with invasive phenotypes observed in histological preparations of the mouse-PDX models, and clinical data on metastasis in the patients, are currently ongoing. In conclusion we provide evidence of the accuracy of the ZTX models in predicting anti-tumor responses to commonly used drugs in NSCLC compared to mouse-PDX models and demonstrate that ZTX models provide a sensitive method for determining metastatic risk and the anti-metastatic efficacy of NSCLC relevant drugs. Citation Format: Zaheer Ali, Malin Vildevall, Anna Nilsson, Julia Schueler, Anna Fahlgren, Lasse DE Jensen. Zebrafish patient tumor-derived xenograft models synergize with mouse-PDX models for understanding variation in anti-cancer drug responses [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6125.

Abstract 4077: Quantitative proteomics reveals novel immunomodulatory pathways of resistance to PARP therapy

Abstract Background Pharmacological inhibition of PARP results in the specific killing of BRCA1/2 deficient tumor cells due to a synthetic lethal interaction between the concomitant impairment in homologous recombination and the DNA damage response. Despite the success of this approach, resistance to PARP inhibition has been observed in majority of patients with advanced cancer. Novel ways of interrogating PARP resistance are necessary to further elucidate the mechanisms of drug resistance and help identify ways to overcome it. To probe potential mechanisms of resistance we applied data-independent acquisition (DIA) mass spectrometry for unbiased global protein quantification in patient derived xenografts with demonstrated resistance to PARP inhibitors. Methods Patient-derived tumor xenografts (PDTXs) were generated from breast cancer patient tumor material implanted in severely immuno-compromised NOD-scid IL2rgnull (NSG) mice. PDTXs, developed as part of the CRUK Cambridge Institute biobank of PDTXs, have previously undergone extensive molecular profiling. PDTXs and short-term cultured PDTX cells (or PDTCs) capture most of originating patient's sample features, including heterogeneity, and consequently, the PDTX/PDTC platform is a robust intermediate in oncogenic drug development. Resistance to the PARP inhibitors AZD-2281 (olaparib) and BMN-673 (talazoparib) was tested ex vivo in PDTCs from 6 sensitive and 11 resistant PDTXs. Deep quantitative proteome profiling of PDTXs samples was conducted by applying an LC-MS/MS setup operated in DIA mode. Data was extracted using Spectronaut™ (Biognosys) with a sample specific spectral library. Pathway analysis was conducted to highlight dysregulated biological functions and pathways and the proteomics data was correlated with transcriptomics readouts. Results LC-MS/MS profiling allowed the identification of more than 11'000 proteins with more than 8'700 proteins quantified across the samples. 448 human and 430 murine proteins were significantly changed between PARP inhibitor resistant and sensitive PDTX models. Resistant models were characterized by increased expression of DNA damage response proteins including ATR and FANCD2, downregulation of TP53, TP53BP1, POLB and H2AFX, and upregulated EGFR, BRAF, ERBB2, STAT3, MLLT4 and CDKN2A. Most interestingly, we observed upregulation of human CD47/SIRPa immunomodulatory signals and upregulation of mouse Ly6G, CSF1R and SHP-1 suggesting the infiltration of immunosuppressive neutrophils and monocytes in the tumor microenvironment in the models resistant to PARP inhibition. Conclusions To our knowledge, this is the first report aiming at interrogating PARP inhibitor resistance with unbiased quantitative proteomics. The proteomics data confirmed prior observations of resistance mechanisms to PARP and elucidated potential novel mechanisms involving modulation of the immune response in resistant tumors. Citation Format: Jakob Vowinckel, Yuehan Feng, Dimitra Georgopoulou, Alejandra Bruna, Tobias Treiber, Abigail Shea, Giulia Lerda, Martin O'Reilly, Kristina Beeler, Carlos Caldas. Quantitative proteomics reveals novel immunomodulatory pathways of resistance to PARP therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4077.

Abstract 3452: Pre-clinical development of next generation selective estrogen receptor degrader - SAR439859

Abstract Abstract: Nearly 70% or more of newly diagnosed cases of breast cancer (BC) are estrogen receptor positive (ER+) where endocrine therapy is a primary treatment. However, substantial evidence describes a continued role of ER signaling in tumor progression, where approximately 40% of patients on endocrine therapy develop resistance that include mutations in ER that drive a constitutively active receptor. SAR439859 is a novel next generation SERD (selective estrogen receptor degrader) with potent antagonist and degradation activity against ER both wild type and mutant and currently evaluated as a single agent and in combination with palbociclib in a Phase I/II clinical trial (NCT03284957). Here we describe the preclinical development of this molecule in multiple xenografts and demonstrated significant anti-tumor efficacy/regression in ER+ BC models including MCF7-ESR1 mutant-Y537S and endocrine therapy resistant patient-derived xenograft tumor models. The molecule also demonstrated strong synergistic activity with CDK4 inhibitor palbociclib. SAR439859 has a favorable safe profile and no uterotrophic activities. Collectively, these results show that SAR439859 is an oral, selective estrogen receptor antagonist and degrader that could provide therapeutic benefit to ER+ breast cancer patients. In conclusion, SAR439859 demonstrates broad anti-tumor activity in ER+ breast cancer cell line/PDX models. Anti-tumor efficacy correlates with PK exposure/PD modulation in target tissue. SAR439859 also has synergistic activity with CDK4 inhibitor Palbociclib. Citation Format: Fangxian Sun, Jane Cheng, Amy Sullivan, Natalia Malkova, Maysoun Shomali, Jesseca McManus, Mikhail Levit, Dinesh Bangari, Youssef El-Ahmad, Sukhvinder Sidhu, Laurent Debussche, Monsif Bouaboula. Pre-clinical development of next generation selective estrogen receptor degrader - SAR439859 [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3452.

Abstract 5634: The single mouse trial format predicts the sensitivity towards checkpoint inhibitor treatment in NSCLC PDX models in HuCD34NCG/CRL mice, an innovative triple immune deficient mouse strain

Abstract Immunotherapy is revolutionizing cancer treatment. As a consequence, there is an urgent need for predictive preclinical models to support compound development with the aim of bringing optimal drug candidates into clinical trials. Patient-derived xenografts (PDX) are the gold-standard in cancer drug development. One major drawback of PDX is the lack of an immunological competent host. To overcome this hurdle triple immunodeficient mice are humanized with hematopoietic stem cells (HSC) to enable the interaction between human immune cells and human tumor cells in a rodent host. The current study uses HSC humanized NCG (NOD-Prkdcem26Cd52Il2rgem26Cd22/NjuCrl) mice, HuCD34NCG, to evaluate the sensitivity of five NSCLC PDX models towards anti CTLA-4 (Ipilimumab), anti-PD-1 (Nivolumab) and the combination thereof. We identified one predicted responder and one non-responder and in a single mouse trial approach (SMT) and confirmed the results in a conventional experimental set-up with n=5 per treatment arm. Infiltration of human immune cells was detected by flow cytometry in tumor tissue (TIL) and hematopoietic organs. Serum of tumor bearing animals was analyzed for human and mouse cytokine secretion pre- and post-treatment. The NCG mouse strain was genetically characterized by whole exome sequencing (WES) and the mutational profile compared to other common mouse strains. The mutational profile showed the highest overlap with the genome data from other triple immune compromised mice and with NOD/SCID indicating their close hereditary relationship. Non-tumor bearing HuCD34NCG mice depicted engraftment of human CD45+ cells in peripheral blood of >25% and >60% in bone marrow and spleen after 8-12 wks post injection. Thus, they fulfilled enrollment criteria for conducting a drug screen for immune modulating compounds. The five NSCLC PDX models showed a distinct sensitivity profile ranging from partial remission (tumor growth inhibition, TGI, of 87.5% compared to untreated control) to progression (no TGI). In all models the combination treatment was the most active therapy (TGI: 60%), followed by Ipilimumab (TGI: 55%) and Nivolumab (TGI: 48%). Nevertheless, in selected models anti-CTLA-4 treatment was the most efficacious therapy. The two models examined in more detail LXFE 397 (=responder, TGI 55% in both formats) and LXFA 400 (non-responder, TGI 2%SMTvs 5%, n=5/group) displayed the same sensitivity pattern in the conventional set-up as compared to the SMT screening approach. The TIL analysis revealed a pronounced rise in the CD3+ subset and increased CD4+/CD8+ ratio in the most effective treatment arms. Subsequent serum analyses will help identify possible biomarkers predicting drug response towards checkpoint inhibition. Taken together our study proves that the HuCD34NCG mouse is a fully functional drug development tool. In combination with highly predictive PDX tumor models this in vivo platform provides a further step to support the development of new drugs targeting the host immune response. Citation Format: Julia Schüler, Jenny Rowe, Eva Oswald, Stephen Festin. The single mouse trial format predicts the sensitivity towards checkpoint inhibitor treatment in NSCLC PDX models in HuCD34NCG/CRL mice, an innovative triple immune deficient mouse strain [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5634.

Abstract 1684: PDX-derived 3D InSightTM immune-competent tumor microtissues as ex-vivo human model for evaluating therapeutic responses

Abstract Background: Current in vitro preclinical models to assess the efficacy of drug candidates only poorly recapitulate the complex features of human tumors. The development of better screening models and platforms comes always with the major question on how accurately and robustly they can represent the complexity of human disease. Patient-derived xenograft (PDX) models faithfully preserve the biological features and the genetic expression profile of human tumor specimens. However, one limiting aspect of patient-derived models is the replacement of the human host microenvironment by murine stroma within the tumor. Lack of cross-species compatibility compromises the induction of a broad range of signaling pathways that cannot be entirely recapitulated. In that respect, our in vitro 3D InSightTM Tumor Microtissues derived from PDX lines retain key properties of the parental human tumors and provide a relevant physiological tumor microenvironment (TME) to assess candidate drugs for novel therapeutic approaches. Aim: Development of in vitro 3D InSightTM Tumor Microtissues from PDX lines aimed to retain the cellular heterogeneity found in the original human tumor tissue. Material & Methods and Results: PDX cell suspensions of lung, breast and melanoma origin were successfully used to assess 3D aggregation in 96-well format and characterized over 10 days in culture. 3D PDX cell cultures were supplied with exogenous normal human dermal fibroblasts (nHDF) and to provide a more physiological cancer niche, PDX cells were also co-cultured with tumor-matched cancer-associated fibroblasts (CAFs). 3D InSight™ Tumor Microtissue from Melanoma PDX were labeled with CellTracker to monitor for tumor cell viability once exposed to peripheral blood mononuclear cells (PBMCs). PBMCs were stimulated with cytokines or anti-CD3/CD28 to generate a pro-inflammatory tumor microenvironment with the subsequent immune-cell mediated tumor attack. To assess the distribution of various cell populations within the tumor, 3D PDX samples were screened for standard stromal vs. epithelial-tumor cells markers (e.g. FAP, pan-CK, E-Cadherin), and diagnostic cancer type-specific biomarkers. 3D PDX samples were also employed to investigate the efficacy of specific targeted therapies based on distinct molecular signatures of PDX tumor models. The gradual modulation of PBMCs mediated either by antibodies or cytokines had a defined impact of the tumor cell killing effect with higher cell death in presence of CD3/CD28 antibodies. Conclusion PDX-derived 3D InSightTM tumor microtissues faithfully model the features and heterogeneity of original human tumor specimens. The successful integration of immune cells (PBMCs) will provide reliable preclinical translational research of tumor/immune cell interactions. We suggest that in vitro 3D PDX models offer a more relevant and robust approach to assess the efficacy of immunomodulators and their combinations to create the new strategies for cancer therapy. Citation Format: Judi Wardwell, Francesca Chiovaro, Nicole Buschmann, Silvan Strebel, Armin Wolf, Irina Agarkova. PDX-derived 3D InSightTM immune-competent tumor microtissues as ex-vivo human model for evaluating therapeutic responses [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1684.

Landscape of G-quadruplex DNA structural regions in breast cancer.

Response and resistance to anticancer therapies vary due to intertumor and intratumor heterogeneity1. Here, we map differentially enriched G-quadruplex (G4) DNA structure-forming regions (∆G4Rs) in 22 breast cancer patient-derived tumor xenograft (PDTX) models. ∆G4Rs are associated with the promoters of highly amplified genes showing high expression, and with somatic single-nucleotide variants. Differences in ΔG4R landscapes reveal seven transcription factor programs across PDTXs. ∆G4R abundance and locations stratify PDTXs into at least three G4-based subtypes. ∆G4Rs in most PDTXs (14 of 22) were found to associate with more than one breast cancer subtype, which we also call an integrative cluster (IC)2. This suggests the frequent coexistence of multiple breast cancer states within a PDTX model, the majority of which display aggressive triple-negative IC10 gene activity. Short-term cultures of PDTX models with increased ∆G4R levels are more sensitive to small molecules targeting G4 DNA. Thus, G4 landscapes reveal additional IC-related intratumor heterogeneity in PDTX biopsies, improving breast cancer stratification and potentially identifying new treatment strategies.

Abstract A24: Establishment of a panel of patient-derived tumor xenograft models recapitulating molecular heterogeneity and drug response of muscle-invasive bladder tumors

Abstract Background: Muscle-invasive bladder cancers (MIBCs) constitute a heterogeneous group of tumors with poor outcome. Recently, MIBC molecular subtyping efforts from an international consortium led to the identification of six subtypes, improving prediction of clinical outcomes and treatment responses. FGFR3 alterations (mutations and translocations), observed in 20% of MIBCs, are found mainly in the luminal papillary subtype that respond poorly to chemo- and immunotherapy. Basal tumors represent 35% of MIBCs and were shown to be better responders to chemotherapy. Here, we describe the development and characterization of patient-derived primary MIBC xenografts (PDX) belonging to these main subtypes. Methods: Bladder tumors were obtained from patients at surgery. Tumor fragments were subcutaneously engrafted into immune-compromised mice. Primary tumors and matched PDX tumors at multiple passages were analyzed regarding growth characteristics, histopathology (H&E staining, CK5/6, FOXA1, and GATA3 immunohistochemistry), gene expression (Affymetrix U133 plus 2.0 microarray), and genetic stability (STR profiling). Hotspot oncogenic mutations for FGFR3, PIK3CA, HRAS, KRAS, NRAS, PPARG, and RXRA were also assessed. Additionally, pharmacologic responses to standard-of-care and targeted therapies were characterized. Findings: From 152 MIBC tumors at all stages and grades, 32 PDX models were successfully established (21.1% success rate). This take rate did not seem correlated to any classical tumor characteristics. Importantly, transcriptomic analysis allowed us to identify PDX models belonging to different molecular subtypes, notably the basal-like and luminal papillary subtypes, including PDXs with FGFR3 mutations. All histologic, genetic, and molecular features validated the stability of the PDX models compared to the parental tumors. Histologic analyses correlated with the molecular classification. These models reproduced the response to cisplatin-based therapies observed in the clinic. Basal models, except one harboring a FGFR3 mutation, were sensitive to anti-EGFR therapies but to a lesser extent than to chemotherapy. FGFR3-mutated PDX models, including a basal model, were highly responsive to FGFR3 inhibitors and less responsive to chemotherapy. Conclusion: We have developed and characterized highly relevant preclinical models for MIBCs, including basal and FGFR3-mutated tumors, recapitulating molecular heterogeneity and drug responses as observed in patients with MIBCs. They represent essential tools for developing new, efficient therapies against this deadly disease. Citation Format: Claire Béraud, Hervé Lang, Myriam Lassalle, Véronique Lindner, Aurélie Kamoun, Michel Soulié, Elodie Guillon, Clémentine Krucker, Xavier Gamé, Pascal Rischmann, Aurélien De Reynies, Yves Allory, François Radvanyi, Philippe Lluel, Thierry Massfelder, Isabelle Bernard-Pierrot. Establishment of a panel of patient-derived tumor xenograft models recapitulating molecular heterogeneity and drug response of muscle-invasive bladder tumors [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2019 May 18-21; Denver, CO. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(15_Suppl):Abstract nr A24.

Patient-Derived Tumor Xenograft Models: Toward the Establishment of Precision Cancer Medicine.

Patient-derived xenografts (PDXs) describe models involving the implantation of patient-derived tumor tissue into immunodeficient mice. Compared with conventional preclinical models involving the implantation of cancer cell lines into mice, PDXs can be characterized by the preservation of tumor heterogeneity, and the tumor microenvironment (including stroma/vasculature) more closely resembles that in patients. Consequently, the use of PDX models has improved the predictability of clinical therapeutic responses to 80% or greater, compared with approximately 5% for existing models. In the future, molecular biological analyses, omics analyses, and other experiments will be conducted using recently prepared PDX models under the strong expectation that the analysis of cancer pathophysiology, stem cells, and novel treatment targets and biomarkers will be improved, thereby promoting drug development. This review outlines the methods for preparing PDX models, advances in cancer research using PDX mice, and perspectives for the establishment of precision cancer medicine within the framework of personalized cancer medicine.

Downregulation of Jumonji-C domain-containing protein 5 inhibits proliferation by silibinin in the oral cancer PDTX model.

Dysregulation of histone demethylase Jumonji-C domain-containing protein 5 (JMJD5) has been identified as a great effect on tumorigenesis. Silibinin is a commonly used anti-hepatotoxic drug and exhibits anticancer effect in various cancers. However, the antitumor mechanism between silibinin and JMJD5 in oral squamous cell carcinoma (OSCC) remains unclear. In this study, the clinical significance of JMJD5 on OSCC patients was assessed through tissue microarray. Furthermore, mice bearing patient-derived tumor xenografts (PDTXs) and tongue cancer cell lines were treated with silibinin and evaluated for tumor growth and JMJD5 expression. High expression of JMJD5 in oral cancer was significantly associated with tumor size (P = 0.0241), cervical node metastasis (P = 0.0001) and clinical stage (P = 0.0002), was associated with worse survival rate compared with that of the total cohort (P = 0.0002). Collectively the data indicate that JMJD5 expression may be suitable for detection of unfavorable prognosis in OSCC patients, based in part on its apparent role as a marker of metastasis. In addition, silibinin inhibits cancer growth in vitro and in PDTX models. Furthermore, metastasis-associated protein 1 (MTA1) could regulate the expression for JMJD5 and had a positive correlation with JMJD5. Moreover, silibinin could downregulate JMJD5 and MTA1 in oral cancer. Present study thus identifies that JMJD5 might be an essential prognostic indicator and therapeutic target against OSCC progression. In addition, silibinin is a potential candidate among novel chemotherapeutic agents or adjuvants for modulating JMJD5 in OSCC, through a mechanism likely involving MTA1/JMJD5 axis.

Abstract A45: Targeted drug therapies for osteosarcoma

Abstract Osteosarcoma (OS) is a highly aggressive cancer that has had no new treatments options in over 30 years. OS is a heterogeneous disease that is characterized by widespread and recurrent somatic copy-number alterations (SCNAs) with relatively few recurrent point mutations. Our laboratory has recently published that SCNAs contain key oncogenic drivers that can be used to identify patient-specific candidates for targeted therapies (Sayles, Breese, et al., Cancer Discovery Jan. 2019). Using patient-derived tumor xenografts (PDX), we demonstrated that targeting of patient-specific oncogenes within SCNAs leads to significant decrease in tumor burden. However, no single-agent therapy was able to regress tumors completely, suggesting that combination therapies would be required for disease management. In order to assess the applicability of our PDX models to the patient tumor and the stability of the oncogenes within SCNAs, we performed whole-genome sequencing (WGS). We observed that SCNAs are highly stable across samples from the same patient in addition to multiple PDX passages and PDX-derived cell lines, highlighting the equivalence between the PDX models and their derived cell lines to the human disease. This allows us to use the PDX cell lines as a surrogate for the identification of combination drug therapies that may be of benefit in OS. Currently, we have generated 6 PDX cell lines that encompass the most common SCNAs observed in patients, including MYC, CCNE1, and CDK4 gain and alterations in the PI3K/PTEN pathway. We have performed a single-agent drug screen with 38 agents and have identified several efficacious compounds, including HDAC inhibitors and cell cycle and Wee1 inhibitors. We also observed a differential response between PDX cell lines to various chemotherapeutics, including gemcitabine, paclitaxel, and SN-38. We are currently testing combination drug therapies in vitro and will validate using our PDX xenografts in vivo. While these experiments are still ongoing, we can report that MYC-driven OS PDX xenografts show a striking resensitization to cisplatin after AURKB inhibitor (barasertib) pretreatment. We have 2 aggressive PDX models with high MYC amplification generated from metastatic lesions. These PDX models are resistant to cisplatin, which is part of the standard of care. We treated these PDX with barasertib and then with cisplatin for two cycles. This resulted in a decrease in tumor volume for the combination therapy compared to vehicle or either single agent alone. Additionally, we observed retention of the platinum adduct 24 hrs after cisplatin dosing only when tumors were pretreated with barasertib. Further work is needed to assess the mechanism of this synergy and whether it can be of use in other SCNA-driven OS and to identify other possible combination therapies that could be of importance in this disease. Citation Format: Leanne C. Sayles, Marcus R. Breese, Henry Martell, Alex G. Lee, Stanley Leung, Avanthi T. Shah, E. Alejandro Sweet-Cordero. Targeted drug therapies for osteosarcoma [abstract]. In: Proceedings of the AACR Special Conference on the Advances in Pediatric Cancer Research; 2019 Sep 17-20; Montreal, QC, Canada. Philadelphia (PA): AACR; Cancer Res 2020;80(14 Suppl):Abstract nr A45.

Enhanced Chemotherapeutic Efficacy of PLGA-Encapsulated Epigallocatechin Gallate (EGCG) Against Human Lung Cancer.

PurposeCurrently, the clinical benefits of tea polyphenols have contributed to the development of efficient systemic delivery systems with adequate bioavailability and stability. In this study, we aimed to establish a nanoparticle model to overcome the shortcomings of epigallocatechin gallate (EGCG) in the treatment of lung cancer.Materials and MethodsPoly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) loaded with EGCG were prepared by the oil-in-water emulsion solvent evaporation technique. The characteristics of NPs, entrapment efficiency, and in vitro release were systematically evaluated. The cellular uptake, cytotoxic activity, and the effect of the formulation on cellular apoptosis of free-from EGCG and the NPs were compared. The interaction between protein-NF-κB and EGCG was detected by bio-layer interferometry (BLI). NF-κB signaling was evaluated by Western blotting and q-RT-PCR. The efficacy of the optimized nanoformulation was evaluated using a patient-derived tumor xenograft (PDX) model.ResultsEGCG-loaded NPs (175.8±3.8 nm in size) demonstrated its optimal efficacy, with approximately 86.0% of encapsulation efficiency and 14.2% of loading efficiency. Additionally, EGCG-encapsulated PLGA-NPs offered a 3-4-fold dose advantage compared to free EGCG in terms of exerting antiproliferative effects and inducing apoptosis at lower doses (12.5, 25 μM). Molecular interaction assays demonstrated that EGCG binds to NF-κB with high affnity (KD=4.8×10−5 M). EGCG-NPs were more effective at inhibiting NF-κB activation and suppressing the expression of NF-κB-regulated genes than free EGCG. Furthermore, EGCG-NPs showed superior anticancer activity in the PDX model than free EGCG.ConclusionThese findings indicated that the prepared EGCG-NPs were more effective than free EGCG in inhibiting lung cancer tumors in the PDX model.

A Novel JAK1 Mutant Breast Implant-Associated Anaplastic Large Cell Lymphoma Patient-Derived Xenograft Fostering Pre-Clinical Discoveries.

Breast implant-associated lymphoma (BIA-ALCL) has recently been recognized as an independent peripheral T-cell lymphoma (PTCL) entity. In this study, we generated the first BIA-ALCL patient-derived tumor xenograft (PDTX) model (IL89) and a matching continuous cell line (IL89_CL#3488) to discover potential vulnerabilities and druggable targets. We characterized IL89 and IL89_CL#3488, both phenotypically and genotypically, and demonstrated that they closely resemble the matching human primary lymphoma. The tumor content underwent significant enrichment along passages, as confirmed by the increased variant allele frequency (VAF) of mutations. Known aberrations (JAK1 and KMT2C) were identified, together with novel hits, including PDGFB, PDGFRA, and SETBP1. A deep sequencing approach allowed the detection of mutations below the Whole Exome Sequencing (WES) sensitivity threshold, including JAK1G1097D, in the primary sample. RNA sequencing confirmed the expression of a signature of differentially expressed genes in BIA-ALCL. Next, we tested IL89’s sensitivity to the JAK inhibitor ruxolitinib and observed a potent anti-tumor effect, both in vitro and in vivo. We also implemented a high-throughput drug screening approach to identify compounds associated with increased responses in the presence of ruxolitinib. In conclusion, these new IL89 BIA-ALCL models closely recapitulate the primary correspondent lymphoma and represent an informative platform for dissecting the molecular features of BIA-ALCL and performing pre-clinical drug discovery studies, fostering the development of new precision medicine approaches.