Establishment Of Patient-derived Xenografts Research Articles

Patient-Derived Xenograft Establishment from Human Malignant Pleural Mesothelioma.

Purpose: Malignant pleural mesothelioma (MPM) is a rare but aggressive disease with few therapeutic options. The tumor-stromal interface is important in MPM, but this is lost in cell lines, the main model used for preclinical studies. We sought to characterize MPM patient-derived xenografts (PDX) to determine their suitability as preclinical models and whether tumors that engraft reflect a more aggressive biological phenotype.Experimental Design: Fresh tumors were harvested from extrapleural pneumonectomy, decortication, or biopsy samples of 50 MPM patients and implanted subcutaneously into immunodeficient mice and serially passaged for up to five generations. We correlated selected mesothelioma biomarkers between PDX and patient tumors, and PDX establishment with the clinical pathologic features of the patients, including their survival. DNA of nine PDXs was profiled using the OncoScan FFPE Express platform. Ten PDXs were treated with cisplatin and pemetrexed.Results: A PDX was formed in 20 of 50 (40%) tumors implanted. Histologically, PDX models closely resembled the parent tumor. PDX models formed despite preoperative chemotherapy and radiotherapy. In multivariable analysis, patients whose tumors formed a PDX had significantly poorer survival when the model was adjusted for preoperative treatment (HR, 2.46; 95% confidence interval, 1.1-5.52; P = 0.028). Among 10 models treated with cisplatin, seven demonstrated growth inhibition. Genomic abnormalities seen in nine PDX models were similar to that previously reported.Conclusions: Patients whose tumors form PDX models have poorer clinical outcomes. MPM PDX tumors closely resemble the genotype and phenotype of parent tumors, making them valuable models for preclinical studies. Clin Cancer Res; 23(4); 1060-7. ©2016 AACR.

24P Establishment of patient-derived xenografts from malignant pleural effusion using NOG mice

Background Personalized medicine represents an ideal medical approach for cancer therapy. Xenografts derived from engrafting fresh surgical specimens directly into immunodeficient mice have recently enabled the development of more relevant in vivo models for human tumors. Patient-derived xenograft (PDX) models retain similar morphologies, heterogeneities, and molecular signatures to the original cancers, and, thus, may be used in the rapid screening of potential therapeutics. We previously reported the rapid and efficient establishment of PDXs using super-immunodeficient NOG mice (PDX/NOG). Surgical specimens are clearly necessary for establishing individual PDX/NOG using personalized medicine. In this study, we attempted to establish PDX/NOG from malignant pleural effusion (MPE) and investigated the potential of MPE-PDX/NOG for personalized anti-cancer therapies. Methods All MPEs were obtained from therapeutic thoracentesis with the informed consent of patients. Cell pellets combined with cancer cells and unarranged cells were generated by centrifuging MPE, and were then inoculated subcutaneously into NOG mice. The established xenograft models were confirmed pathologically after tumor tissue had been passaged three times by subcutaneous engraftment. The preservation of cancer stem cells (CSCs) was confirmed by immunohistochemical and gene expression analyses. Results We established 2 MPE-PDX/NOG lines (pulmonary adenocarcinoma) from 4 cases. These lines showed the histological structures of well-differentiated adenocarcinoma with murine stromal formation. The morphological characteristics of MPE-PDX/NOG were maintained for cellularity and structural heterogeneity. CSC markers (CD44, ALDH1A1, and EpCAM) were preserved in MPE-PDX/NOG tissue. Conclusions The establishment of PDX/NOG from MPE is efficient and as valuable as that from surgical specimens. Serial MPE-PDX/NOGs may be less invasively established from the same patient because patients with MPE may repeatedly undergo therapeutic thoracentesis. The analysis of serial MPE-PDX/NOGs will disclose characteristic changes in cancer cells affected by chemotherapies and contribute to personalized medicine. Legal entity responsible for the study Jichi Medical University Funding JSPS Grant-in-Aid for Scientific Research Disclosure All authors have declared no conflicts of interest.

24P Establishment of patient-derived xenografts from malignant pleural effusion using NOG mice

24P Establishment of patient-derived xenografts from malignant pleural effusion using NOG mice

Abstract B22: Establishment of Patient-Derived Xenograft (PDX) Models of Ovarian Cancer

Abstract Ovarian cancer is the most lethal gynecological malignancy. The majority of ovarian cancer patients are diagnosed at an advanced stage of disease, and the lack of good early screening and diagnostic tests is one of the reasons for the low overall survival rate. Moreover, there is no good marker to predict patients' response to chemotherapy. Patient-derived xenografts (PDXs) have become the most promising preclinical model for gynecologic malignances. We have successfully established 22 PDX models derived from ovarian cancer patients' surgical specimens implanted into severe combined immunodeficient (SCID) mice, non-obese diabetic (NOD)-SCID or Nude mice. The majority of the PDX models were generated from patients' samples diagnosed with high-grade serous carcinoma including 3 with BRCA1/2 mutations, as well as less common types of ovarian cancers such as low-grade serous, endometrioid, mucinous carcinomas, and carcinosarcoma. PDXs have been established from solid masses as well as from ascites. We have optimized protocol for establishing ovarian PDXs including implantation and collection methods, tumor type (solid versus ascites), and type of mouse strains used. Using SCID mice, our success rate exceeded 80% and recent data indicated that the success rate in NOD/SCID mice was higher. The link between the pathology and PDX core, the “triage protocol” and ongoing PARP inhibitor trials will provide an important array of PDXs including pre- and post-treatment PDXs for exploration for biomarkers as well as for co-clinical trials to validate the utility of rational drug combinations. All models were validated using short tandem repeats (STR) and compared to the H&E and immunohistochemistry stains from the matching patient's tissue. We also examined the expression of B7-H4, a novel member of the B7 family immune checkpoint regulators, in 9 patient-derived xenografts (PDXs) of high-grade ovarian serous carcinoma. Our data showed that B7-H4 proteins were expressed in 8 of 9 PDX models of high-grade serous carcinoma, suggesting that PDXs may be useful for studying immunotherapy. Citation Format: Li Liang, Imelda Mercado-Uribe, Na Niu, Yi Jiang, Wenjun Cheng, Russell Broaddus, Robert Bast, Gordon Mills, Anil K. Sood, Jinsong Liu. Establishment of Patient-Derived Xenograft (PDX) Models of Ovarian Cancer. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr B22.

Abstract B38: Clinical applications of PDX/NOG models for personalized chemotherapy – possible use in chemo-sensitivity testing and clinical sequencing

Abstract Personalized medicine represents an ideal medical approach for cancer therapy. In the field of clinical oncology, personalized medicine or therapy involves the evolutionary expansion of conventional clinical approaches that progress from patient evaluation, differential diagnosis, to the treatment of diseases. A number of the complex techniques employed in personalized medicine, such as clinical genome sequencing, biochemical marker analyses, chemo-sensitivity testing, and cancer immunotherapy, require fresh, viable, and sufficient amounts of specimens for reliable estimations. Mice bearing patient-derived xenografts (PDXs) with clinical information (so-called “Cancer Xenopatients”) are remarkable systems in personalized medicine for cancer. We previously reported the rapid and efficient establishment of PDXs using NOG mice (PDX/NOG models, AACR2015 #1940, IJO 47 61-70 2015). NOG mice, NOD/Shi-scid/IL2Rγnull (NOG) mice derived from NOD/SCID mice with a common gamma chain, have multifunctional defects in natural killer cell activity, macrophage function, complement activity, and dendritic cell function in addition to the absence of functional T and B lymphocytes. NOG mice have been identified as the most appropriate immunodeficient host animal for the direct xenografting of fresh tumor tissue due to the preservation of cancer stem cells (CSCs). Fresh and valuable xenograft samples, similar to surgical samples with the preservation of CSC, are stably provided using PDX/NOG models. Moreover, human tissue (tumor) and mouse tissue (stroma) are clearly distinguished by immunohistochemical analysis or gene arrangement sequencing. In the present study, we discussed the possibility of using PDX/NOG model simulations for personalized cancer chemotherapy. We previously established 47 lines of gastrointestinal cancer xenografts. In these cases, clinical information regarding chemotherapy for donor patients was retrieved where possible. Collagen gel droplet-embedded culture-drug sensitivity tests (CD-DST) were performed on 16 lines of PDX/NOG. In 4 of these lines, CD-DST were successfully conducted on original surgical specimens. The results of CD-DST between original and PDX/NOG specimens generally correlated (R2=0.01-0.89). The results of CD-DST using PDX/NOG specimens were compatible with the clinical effects of anti-cancer drugs. Genome sequencing and interactome analyses, a comprehensive analysis of tumor-stroma interactions innovated by Professor Ishikawa S. at Tokyo Medical and Dental University, were also performed on mRNA from 17 lines of PDX/NOG. Our interactome analyses showed tumor-stroma interactions in PDX/NOG comprehensively and quantitatively at the gene-expression level by distinguishing gene arrangements in human tissue (tumor) from mouse tissue (stroma). The EGF-EGFR or VEGFA-KDR interactions observed closely reflected the clinical effectiveness of an EGFR inhibitor (Cetuximab) or VEGF-A inhibitor (Bevacizumab) as well as the results of in vivo chemo-sensitivity tests using PDX/NOG. The results of CD-DST and sequencing in PDX/NOG appear to be reliable for clinical simulations of chemotherapy and will definitely assist in the selection of the most sensitive anti-cancer drug for each patient. The fast and efficient establishment of individual PDXs will contribute to personalized anti-cancer therapies. Citation Format: Tsuyoshi Chijiwa, Takayuki Isagawa, Akira Noguchi, Hidemitsu Sato, Akimune Hayashi, Haruhiko Cho, Manabu Shiozawa, Takeshi Kishida, Soichiro Morinaga, Tomoyuki Yokose, Makoto Katayama, Nobuo Takenaka, Mizuha Haraguchi, Naoki Miyao, Yuichi Tateishi, Kenji Kawai, Hiroshi Suemizu, Roppei Yamada, Yoshiyasu Nakamura, Kohzoh Imai, Daisuke Komura, Shumpei Ishikawa, Masato Nakamura, Yohei Miyagi. Clinical applications of PDX/NOG models for personalized chemotherapy – possible use in chemo-sensitivity testing and clinical sequencing. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr B38.

Abstract 639: Subtyping of pancreatic cancer patient-derived xenograft tumors and implications for anticancer agent testing

Abstract Despite improvements in treatment, pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal cancers, with a continuous increase in incidence emphasizing the need for further research and therapeutic development. In recent years, we have developed a collection of >40 patient-derived xenograft (PDX) models from PDAC. In order to determine their relevance for anticancer agent testing, we extensively characterized our models for histology features, whole exome mutations (Hiseq 2000), chromosome rearrangements, gene copy number variations (Affymetrix SNP6) and gene expression (Affymetrix U133 Plus2.0). PDAC from 65 patients were implanted into immuno-compromised mice, resulting in the development of 42 PDX models (success rate 65%). The PDAC from which models were established included moderate and poorly differentiated tumors and were heterogeneous for stroma content. In patient tumors, we showed fibroblast activation but not stroma content predicted poor patient outcome (p<0.0001) and success of PDX establishment (p<0.01). Correlating with those of the parental tumors, the resulting PDAC_PDX also showed heterogeneity for stroma content and for murine fibroblast activation. As seen in patient tumors, the PDAC_PDX were characterized by frequent chromosomal instability, with 38% of the models presenting a moderate hypoploidy while the 62% remaining ones showed a hyperploidy. Two models showed hypermutation due to mismatch repair deficiency while the others had on average a lower mutation load when compared to other histotypes such as colon, lung or melanoma tumors. In total, we identified more than 9000 genes altered by homozygous deletions, high gene amplifications and/or mutations, most of the models showed alterations in KRAS (81%), TP53 (67%), CDKN2A (64%) and TGFBR2/SMAD4 (64%) genes. Next, PDAC_PDX profiles were merged with those of patient tumors to determine PDAC_PDX subtypes and were found to be of the classical (61%), quasi mesenchymal (QM) (34%) and exocrine like subtype (5%) according to the 62 gene expression signature established by Collisson et al. 2011. Preliminary biomarker analysis revealed certain associations between genomic alterations and transcriptome subtypes, such as more frequent alterations in the TGFB pathway in classical PDX (80%) compared to QM (47%). Further detailed biomarkers analyses also addressing questions of molecular determinants of stroma content, of fibroblast activation and of sensitivity toward anticancer agents will be presented. Extensive characterization of our PDAC_PDX collection revealed similarities with patient tumors with regards of histology features including stroma content and fibroblast activation. At molecular level, the models showed similar genomic and transcriptomic patterns as those reported for patient PDAC, altogether, proving the value of this collection for drug development investigations. Citation Format: Peter Bronsert, Tim Kees, Bruno Zeitouni, Anne-Lise Peille, Manuel Landesfeind, Heinz-Herbert Fiebig, Simon Kuesters, Vincent Vuaroqueaux. Subtyping of pancreatic cancer patient-derived xenograft tumors and implications for anticancer agent testing. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 639.

Comprehensive characterization of the patient-derived xenograft and the paralleled primary hepatocellular carcinoma cell line.

BackgroundHepatocellular carcinoma (HCC) is an aggressive cancer with high mortality and morbidity worldwide. The limited clinically relevant model has impeded the development of effective HCC treatment strategy. Patient-derived xenograft (PDX) models retain most of the characteristics of original tumors and were shown to be highly predictive for clinical outcomes. Notably, primary cell line models allow in-depth molecular characterization and high-throughput analysis. Combined usage of the two models would provide an excellent tool for systematic study of therapeutic strategies. Here, we comprehensively characterized the novel PDX and the paralleled primary HCC cell line model.MethodsTumor tissues were collected from HCC surgical specimens. HCC cells were sorted for in vivo PDX and in vitro cell line establishment by the expression of hepatic cancer stem cell marker to enhance cell viability and the rate of success on subsequent culture. The PDX and its matching primary cell line were authenticated and characterized in vitro and in vivo.ResultsAmong the successful cases for generating PDXs and primary cells, HCC40 is capable for both PDX and primary cell line establishment, which were then further characterized. The novel HCC40-PDX and HCC40-CL exhibited consistent phenotypic characteristics as the original tumor in terms of HBV protein and AFP expressions. In common with HCC40-PDX, HCC40-CL was tumorigenic in immunocompromised mice. The migration ability in vitro and metastatic properties in vivo echoed the clinical feature of venous infiltration. Genetic profiling by short tandem repeat analysis and p53 mutation pattern consolidated that both the HCC40-PDX and HCC40-CL models were derived from the HCC40 clinical specimen.ConclusionsThe paralleled establishment of PDX and primary cell line would serve as useful models in comprehensive studies for HCC pathogenesis and therapeutics development for personalized treatment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12935-016-0322-5) contains supplementary material, which is available to authorized users.

Establishment of Patient-derived Xenografts in Mice.

Patient-derived xenograft (PDX) models for cancer research have recently attracted considerable attention in both the academy and industry (Hidalgo et al., 2014; Wilding and Bodmer, 2014). PDX models have been developed from different tumor types including lung cancer to improve the drug development process. These models are used for pre-clinical drug evaluation and can be used for the predictive results of clinical outcomes because they conserve original tumor characteristics such as heterogeneity, complexity and molecular diversity (Kopetz et al., 2012). Additionally, PDX model provides the potential tool for the personalized drug therapy. In this protocol, we present methods for the establishment of PDX in mice using primary tumor tissues from patients with small cell lung cancer (SCLC).

Abstract 1449: The valuable experimental model system “Cancer Xenopatient” established in NOG mice

Abstract Viable and stable human cancer materials combined with adequate clinical information are required for advance in cancer research and patient care. Conventional in vitro cancer cell lines are commonly available without enough information on the patient including disease phenotypes and drug-sensitivity. Mice bearing Patient-derived xenografts (PDXs) with clinical information (so-called “cancer xenopatients”) could be essential and useful systems to accelerate cancer medicine. In this study, we investigated the establishment efficiency of PDX using NOG mice with clinical factors of xeno-transplantaion. The NOG mouse, the NOD/Shi-scid/IL-2Rγnull (NOG) mice derived from the NOD/SCID mouse with a common gamma chain, has multifunctional defects in natural killer cell activity, macrophage function, complement activity, and dendritic cell function in addition to lacking functional T and B lymphocytes. NOG mice were reported to be the best appropriate immunodeficient host animal for direct xenografting of fresh tumor tissue. We also discuss herein its contribution for not only reliable preclinical studies of new anticancer drugs but also personalized anti-cancer therapies. Sixty-one PDX lines were established from 116 surgically removed tumor tissues inoculated into NOG mice's subcutaneous (53%). PDX lines were established from various types of epithelial tumors and also from sarcomas including gastrointestinal stromal tumor and Ewing/PNET sarcoma. The metastatic tumors yielded PDX lines effectively (65%) than the primary tumors (27%, p<0.001). The group engrafted into NOG mice after 2 days or later from the surgical removal showed a higher establishment rate (61%) than that of the group engrafted at the day of surgery or the next day (51%), however non-significant (p = 0.49). Our PDX models were preserved well not only in morphological characteristics but also in gene expression and alteration patterns. In 8 cases (7%), the transplantable xenografts for several generations were composed of monotonous large non-epithelial cell growth of human origin, revealed to be the Epstein-Barr virus (EBV) infection associated lymphoprolipherative lesions prospected by chromogenic in situ hybridization for EBV-encoded RNA. We revealed efficient establishment rates both for primary tumors and metastatic tumors. PDXs linked with clinical information will contribute to reliable preclinical studies for new anticancer drugs. The fast and efficient establishment of individual PDXs may also contribute to future personalized anti-cancer therapies. Citation Format: Tsuyoshi Chijiwa, Kenji Kawai, Akira Noguchi, Hidemitsu Sato, Akimune Hayashi, Haruhiko Cho, Manabu Shiozawa, Takeshi Kishida, Soichiro Morinaga, Tomoyuki Yokose, Makoto Katayama, Nobuo Takenaka, Hiroshi Suemizu, Roppei Yamada, Yoshiyasu Nakamura, Yasuo Takano, Kohzoh Imai, Yohei Miyagi, Masato Nakamura. The valuable experimental model system “Cancer Xenopatient” established in NOG mice. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1449. doi:10.1158/1538-7445.AM2015-1449

Abstract 5131: Establishment of uterine carcinosarcoma primary cell lines for chemosensitivity testing and evaluation of targeted therapy

Abstract To establish primary cell lines from uterine carcinosarcoma (CS) patient samples and to evaluate combination treatment with standard chemotherapy with or without the addition of an anti-IGF1R antibody. CS tumor tissue was obtained under IRB approval at the time of primary surgery. We reported the establishment of patient-derived xenografts elsewhere. Tissue pieces were minced extensively and digested by collagenase treatment. The single cell solution was then seeded in F-media containing EGF, insulin, hydrocortisone, adenine, cholera toxin and the Rock inhibitor Y-27632. A previously established carcinosarcoma cell line CS99 (Schulten et al, 2008) was used for comparison. To verify origin, Short Tandem Repeat (STR) profiles of the primary cell lines CS13, CS18, CS19, CS21 and CS22 were compared with the patient's tumor samples. Drug sensitivity to Taxol, cisplatin and carboplatin was determined individually or in combination using the sulforhodamine B (SRB) proliferation assay. The effect of IMC-A12 (anti-IGF1R antibody, Eli Lily / Imclone) alone or with chemotherapy was also determined with the SRB assay. Total and promoter-specific IGF2 mRNA levels were determined by reverse transcriptase quantitative PCR. All STR profiles of the primary cell lines matched their patient sample counterparts. IGF2 mRNA expression levels of the cell lines were similar to the patient samples, and over 10,000-fold higher than in CS99. IGF2 promoter-specific primers showed that the IGF2 mRNA transcripts of the primary cell lines were initiated at the oncofetal IGF2 promoters P3 and P4. For Taxol, the IC50 (inhibitory concentration of 50%) ranged from 4.7 nM for CS21 to 9.8 nM for CS22. For cisplatin, the IC50 ranged from 1.0 μM for CS99 to 4.8 μM for CS19. For carboplatin, CS99 cells had the lowest IC50 of 12.5 μM while CS19 had the highest IC50 of 58.1 μM. In the presence of 10 μg/ml IMC-A12, the primary cell lines CS19, CS21 and CS22 showed significant sensitization to Taxol (∼50% decrease in IC50), to carboplatin (∼65% decrease in IC50), and to combination Taxol/carboplatin treatment (∼50% decrease in IC50). We conclude that establishing primary cell lines of this rare cancer is a promising approach for testing sensitivity to current and novel chemotherapies and combination treatment strategies. We also found that high oncofetal promoter-driven IGF2 mRNA expression was observed in all CS samples tested and that IGF1R blockade sensitized the primary cell lines to standard chemotherapy. Citation Format: Jurriaan Brouwer-Visser, Eirwen Scott, Shijun Mi, Maria J. Cossio, Tiffany Hebert, Gloria S. Huang. Establishment of uterine carcinosarcoma primary cell lines for chemosensitivity testing and evaluation of targeted therapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5131. doi:10.1158/1538-7445.AM2015-5131

Genomic characterization of a large panel of patient-derived hepatocellular carcinoma xenograft tumor models for preclinical development.

Lack of clinically relevant tumor models dramatically hampers development of effective therapies for hepatocellular carcinoma (HCC). Establishment of patient-derived xenograft (PDX) models that faithfully recapitulate the genetic and phenotypic features of HCC becomes important. In this study, we first established a cohort of 65 stable PDX models of HCC from corresponding Chinese patients. Then we showed that the histology and gene expression patterns of PDX models were highly consistent between xenografts and case-matched original tumors. Genetic alterations, including mutations and DNA copy number alterations (CNAs), of the xenografts correlated well with the published data of HCC patient specimens. Furthermore, differential responses to sorafenib, the standard-of-care agent, in randomly chosen xenografts were unveiled. Finally, in the models expressing high levels of FGFR1 gene according to the genomic data, FGFR1 inhibitor lenvatinib showed greater efficacy than sorafenib. Taken together, our data indicate that PDX models resemble histopathological and genomic characteristics of clinical HCC tumors, as well as recapitulate the differential responses of HCC patients to the standard-of-care treatment. Overall, this large collection of PDX models becomes a clinically relevant platform for drug screening, biomarker discovery and translational research in preclinical setting.

Establishment of patient-derived cancer xenografts in immunodeficient NOG mice.

Viable and stable human cancer cell lines and animal models combined with adequate clinical information are essential for future advances in cancer research and patient care. Conventional in vitro cancer cell lines are commonly available; however, they lack detailed information on the patient from which they originate, including disease phenotype and drug sensitivity. Patient-derived xenografts (PDX) with clinical information (so-called ‘cancer xenopatients’) are a promising advance that may accelerate the development of anticancer therapies. We established 61 PDX lines from 116 surgically removed tumor tissues inoculated subcutaneously into NOG mice (53% success rate). PDX lines were established from various types of epithelial tumors and also from sarcomas, including gastrointestinal stromal tumors and Ewing/PNET sarcomas. The metastatic tumors yielded PDX lines more effectively (65%) than the primary tumors (27%, P<0.001). In our PDX models, morphological characteristics, gene expression profiles, and genetic alteration patterns were all well preserved. In eight cases (7%), the transplantable xenografts for several generations were composed of large monotonous nonepithelial cells of human origin, revealed to be Epstein-Barr virus infection-associated lymphoproliferative lesions. Despite this, PDX linked with clinical information offer many advantages for preclinical studies investigating new anticancer drugs. The fast and efficient establishment of individual PDX may also contribute to future personalized anticancer therapies.

Patient-Derived Xenografts from Non–Small Cell Lung Cancer Brain Metastases Are Valuable Translational Platforms for the Development of Personalized Targeted Therapy

The increasing prevalence of distant metastases from non-small cell lung cancer (NSCLC) indicates an urgent need for novel therapeutic modalities. Brain metastasis is particularly common in NSCLC, with severe adverse effects on clinical prognosis. Although the molecular heterogeneity of NSCLC and availability of various targeted agents suggest personalized therapeutic approaches for such brain metastases, further development of appropriate preclinical models is needed to validate the strategies. We established patient-derived xenografts (PDX) using NSCLC brain metastasis surgical samples and elucidated their possible preclinical and clinical implications for personalized treatment. NSCLC brain metastases (n = 34) showed a significantly higher successful PDX establishment rate than primary specimens (n = 64; 74% vs. 23%). PDXs derived from NSCLC brain metastases recapitulated the pathologic, genetic, and functional properties of corresponding parental tumors. Furthermore, tumor spheres established in vitro from the xenografts under serum-free conditions maintained their in vivo brain metastatic potential. Differential phenotypic and molecular responses to 20 targeted agents could subsequently be screened in vitro using these NSCLC PDXs derived from brain metastases. Although PDX establishment from primary NSCLCs was significantly influenced by histologic subtype, clinical aggressiveness, and genetic alteration status, the brain metastases exhibited consistently adequate in vivo tumor take rate and in vitro tumor sphere formation capacity, regardless of clinical and molecular conditions. Therefore, PDXs from NSCLC brain metastases may better represent the heterogeneous advanced NSCLC population and could be utilized as preclinical models to meet unmet clinical needs such as drug screening for personalized treatments.

A phase II study of afatinib in patients (pts) with metastatic human epidermal growth factor receptor (HER2)-positive trastuzumab refractory esophagogastric (EG) cancer.

59 Background: Trastuzumab combined with chemotherapy is the standard of care for pts with HER2+ EG cancer. Resistance to trastuzumab clearly emerges in this population. Afatinib, a potent ErbB Family Blocker, induced tumor regression in MSKCC HER2+ patient derived xenografts (PDX). This study assesses safety and preliminary efficacy of afatinib in patients with trastuzumab refractory EG cancer. Methods: Pts with HER2+ (IHC 3+ or FISH&gt;2.0) EG adenocarcinoma, after progression on trastuzumab, received oral afatinib 40 mg daily. Archival pre-trastuzumab tissue, tumor biopsy after progression on trastuzumab and after 1 week on afatinib is mandated on protocol for next generation sequencing (NGS), proteomics and establishment of PDX. The primary endpoint-overall clinical benefit at 4 months: stable disease (SD) or partial response (PR). Results: 20 pts treated with afatinib; median age 61, KPS 80; median 2 (1 to 4) prior trastuzumab regimens, 67% of tumors IHC3+; 33% IHC2+/FISH&gt;2.0. Common adverse events included: rash or dry skin (Grade 1/2:80%), diarrhea (Grade 1/2:60%), nausea/vomiting (grade 1/2:40%) fatigue (grade1/2: 25%). To date, 19 pts evaluable for response, 2 PRs and 6 SD, 42% disease stabilization rate (PR+SD) at 4months (4 to 13 mos). PDXs established from biopsies of 7 pts. EGFR amplification was detected in the tumor of 2 pts with PRs and 1 of 6 pts with SD. Recurrent PIK3CA, ERBB3 and MTORmutations were observed. Conclusions: Afatinib shows clinical efficacy and is well tolerated in patients with trastuzumab refractory, heavily pretreated EG cancer. Enrollment has now begun in an afatinib + trastuzumab cohort. Efforts to elucidate the mechanisms of trastuzumab resistance including validation of potential drivers of trastuzumab resistance using HER2+ PDXs are ongoing. Updated molecular and clinical data will be presented. Clinical trial information: NCT01522768.

146 Establishment of patient-derived xenografts (PDX) models for triple negative breast cancer (TNBC) as a pre-clinical platform for drug development

146 Establishment of patient-derived xenografts (PDX) models for triple negative breast cancer (TNBC) as a pre-clinical platform for drug development

178 Establishment of patient-derived xenografts (PDX) models for small cell lung (SCL) as a pre-clinical platform for drug development

178 Establishment of patient-derived xenografts (PDX) models for small cell lung (SCL) as a pre-clinical platform for drug development

Abstract 1189: Establishment of patient-derived xenograft (PDX) models for small cell lung cancer (SCLC) as a preclinical platform for drug development

Abstract Small cell lung cancer comprises 15-20% of all lung cancer cases, and is more invasive and has a higher rate of proliferation with respect to non-small cell lung cancer, leading to a higher mortality rate. Most cases are responsive to chemotherapy, however there is a high rate of recurrence with treated patients and those in advanced stage of the disease often have a refractory response to treatment. As such, the need for new treatment modalities is critical. Use of patient-derived tumor xenograft (PDX) models as clinically-relevant preclinical model for novel drug development has gained widespread adoption in recent years. In previous work, we have reported using the Molecular Response tumor bank, comprising &amp;gt;144,000 viable tumor specimens, to establish more than 100 PDX models of various cancer types. In this work, we describe the development of 8 new SCLC PDX models established from a collection of &amp;gt;300 small cell lung cancer specimens, many of which come from prior-treated and metastatic patients. We report a comprehensive characterization of these models, including: histopathology, immunohistochemistry, and mutation analysis by next-generation sequencing. Additionally, we have evaluated functional response of these models with in vivo pharmacology studies. Current studies are underway to derive correlations between in vivo drug response and mutational status of these models. Our data strongly suggests the potential to use these unique PDX models to aid in efforts in drug development efforts in oncology. Citation Format: Patrick Carlson, Jill Ricono, Chelsea Mullins, Thomas Broudy, Cyrus Mirsaidi, Praveen Nair. Establishment of patient-derived xenograft (PDX) models for small cell lung cancer (SCLC) as a preclinical platform for drug development. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1189. doi:10.1158/1538-7445.AM2014-1189

Establishment of Patient-Derived Xenograft (PDX) Models of Human Breast Cancer.

Patient-derived xenograft (PDX) models of human breast cancer are proving useful for preclinical evaluation of experimental therapeutics. However, until recently, generation of PDX models reflecting the full spectrum of human breast cancers has been an elusive goal. We recently developed a method for establishing serially transplantable, phenotypically stable, human breast cancer xenograft models in immunocompromised mice with comparatively high efficiency (overall ∼25%). These xenografts represent the major clinically defined subtypes of breast cancer [e.g. estrogen receptor positive (ER+), HER2 positive (HER2+), and "triple negative" (TN) breast cancers]. This method, and methods being developed in other laboratories, may soon allow for conducting "animal clinical trials" once sufficient numbers of clinically relevant models are generated. Curr. Protoc. Mouse Biol. 3:21-29 © 2013 by John Wiley & Sons, Inc.