bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A36: Establishment of patient-derived xenograft models of lung adenocarcinoma with two different EGFR mutations, L858R and exon19 deletion
Authors: Kita Kenji , Fukuda Koji , Nishiyama Akihiro , Arai Sachiko , Takeuchi Shinji , Ohtsubo Koushiro , Yamashita Kaname , Tamura Masaya , Matsumoto Isao , Yano Seiji
Published: February 2019

Preclinical tumor models that resemble the characteristics of patients’ tumors are essential for development of novel therapy. Patient-derived tumor xenograft (PDX) models are considered to be interesting candidates. However, success rate of PDX models of lung cancer, especially adenocarcinoma, is much lower than that of other types of tumors, including colon cancer.

This study was conducted to establish clinically relevant PDX models of lung cancer. We obtained fresh surgically resected specimens from 20 NSCLC patients (10 squamous cell carcinomas, 9 adenocarcinomas, and 1 large cell carcinoma) who had submitted the written informed consent for this study, which was performed in accordance with protocols approved by the IRB of Kanazawa University. We inoculated the tumor fragments subcutaneously into immunodeficient mice, NOG mice or SCID hairless outbred (SHO) mice. The tumor take rate was 40% (8/20), with higher rates for squamous cell carcinoma (60%, 6/10) than for adenocarcinoma (22%, 2/9). Interestingly, both of two adenocarcinomas that successfully developed PDX tumors had EGFR mutations, L858R and exon 19 deletion, respectively. These PDX tumors maintained histology of adenocarcinoma and EGFR mutation even after repeated in vivo passages. Furthermore, the third-generation EGFR tyrosine kinase inhibitor (EGFR-TKI) osimertinib dramatically regressed tumors in these EGFR mutated PDX models. Osimertinib treatment is ongoing in these EGFR-mutated PDX models to obtain tumors that acquired resistance to osimertinib.

Our PDX models of lung adenocarcinoma with different EGFR mutations may be useful for investigating the mechanisms of resistance to EGFR-TKIs, including third-generation inhibitor osimertinib.

Citation Format: Kenji Kita, Koji Fukuda, Akihiro Nishiyama, Sachiko Arai, Shinji Takeuchi, Koushiro Ohtsubo, Kaname Yamashita, Masaya Tamura, Isao Matsumoto, Seiji Yano. Establishment of patient-derived xenograft models of lung adenocarcinoma with two different EGFR mutations, L858R and exon19 deletion [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr A36.
Abstract B15: Increased presence of T follicular helper cells in lung adenocarcinoma is associated with mutational load
Authors: Enfield Katey S.S. , Ng Kevin W. , Marshall Erin A. , Martin Spencer D. , Lam Wan L.
Published: February 2019

Tertiary lymphoid organs are ectopic lymphoid formations found in inflamed tissues such as tumors, and their presence has been associated with improved patient outcome. T follicular helper cells (Tfh) reside in the germinal centre of tertiary lymphoid organs, and are required for the maturation of B cells and subsequent antibody response. Whereas the prognostic value of Tfh has been described in breast and colon tumors, they remain uncharacterized in lung tumors. We hypothesize that Tfh cells reside in lung adenocarcinomas and are associated with an increased immune response due to neoantigen exposure.

Gene expression profiles were obtained from 83 paired lung adenocarcinomas and nonmalignant lung tissues from the BC Cancer Agency, and 571 unpaired samples from The Cancer Genome Atlas. Relative proportions of 22 immune cell subsets were inferred from gene expression data using CIBERSORT, a deconvolution algorithm. Identification of tertiary lymphoid organs was achieved through multicolor immunohistochemistry (IHC) staining for T- and B-cell lineage markers (CD3 and CD79a) using whole tissue sections. Proportions of Tfh cells were correlated with tumor mutation load defined as non-silent mutations per megabase (Mann Whitney U test) and patient outcome (Cox proportional hazard model).

The proportion of Tfh cells was significantly increased in tumor tissue compared to nonmalignant lung in both cohorts. We also observed concomitant upregulation of Tfh markers PD1 and CXCR5. Multicolor IHC validated the presence of tertiary lymphoid organs in 19 out of 20 cases assessed. Intriguingly, the increase in the proportion of Tfh cells revealed by CIBERSORT was observed across all disease stages and was validated in an additional cohort of Stage I lung adenocarcinomas. The relative proportion of Tfh cells did, however, increase with increasing tumor mutation burden, suggesting their involvement in an active immune response against tumor neoantigens.

Tfh recruitment appears to be an early event in lung tumor progression and a function of neoantigen exposure, suggesting involvement in an active antitumor response rather than a passive result of chronic inflammation. Further investigation into Tfh in lung adenocarcinomas may lead to prognostic applications.

Citation Format: Katey S.S. Enfield, Kevin W. Ng, Erin A. Marshall, Spencer D. Martin, Wan L. Lam. Increased presence of T follicular helper cells in lung adenocarcinoma is associated with mutational load [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr B15.
Abstract B17: Interleukin-17 and -22 expression in non-small cell lung cancer
Authors: Stump Julia , Jiang Yue , Karches Clara , Gosálvez Javier Suárez , Neumann Jens , Kobold Sebastian , Hatz Rudolf , Winter Hauke , Tufman Amanda , Huber Rudolf M. , Reu Simone
Published: February 2019

Background: In non-small cell lung cancer (NSCLC) the TNM staging remains standard for prognostic assessment and therapy decisions. Nevertheless, stage-specific outcomes vary significantly, indicating a need for additional prognosticators. In this retrospective study we wanted to assess the role of the mediator interleukin-22 (IL-22), as IL-22 is known to be involved in various lung diseases and to be elevated in lung cancer patients. Moreover, we evaluated interleukin-17 (IL-17), which also depicts a proinflammatory cytokine, but seems to play a dual role in antitumor immunity. We asked the question of their link to prognosis, therapy response and recurrence.

Methods: Tissue microarrays (TMAs) were generated from formalin-fixed, paraffin embedded tissue of 138 curatively resected patients with stage IA-IV NSCLC. TMAs included each three cores from the tumor center (CT) and invasive margin (IM), selected from areas with the most dense lymphocytic infiltrates. IL-22 and IL-17 expression of the tissue was analyzed by immunohistochemistry via cytoplasmic staining (double-staining: IL-22, CD3 resp. IL-17, CD3).

Results: IL-22 expression status in adeno and SCC does not seem to have a prognostic impact on survival, regardless of the localization. However, a high IL-22 CT/IM ratio in adenocarcinoma is clearly linked to longer overall survival; this cannot be seen in SCC. For IL-17 a distinct tendency is visible for adeno and SCC in the invasive margin as well as in the tumor center—interestingly, this is more clear for SCC. In SCC a high CT/IM ratio for IL-17 seems to have a positive prognostic impact on overall survival. Patient numbers will be enlarged to validate these findings.

Conclusion: The cytokines IL-22 and IL-17 seem—depending on histology and location—to have a prognostic impact on overall survival in NSCLC. In previous experiments we showed that multispectral assessment of CD8 and PD-L1 has a clear correlation with clinical outcome. Adding cytokines to this method might open novel avenues for predicting clinical outcome and therapeutic efficacy in NSCLC.

Citation Format: Julia Stump, Yue Jiang, Clara Karches, Javier Suárez Gosálvez, Jens Neumann, Sebastian Kobold, Rudolf Hatz, Hauke Winter, Amanda Tufman, Rudolf M. Huber, Simone Reu. Interleukin-17 and -22 expression in non-small cell lung cancer [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr B17.
Abstract B21: Investigating targeted driver mutations and PD-L1 expression for improved therapy of non-small cell lung cancer
Authors: Alwithenani Akram , Forsythe Marika , Castonguay Mathieu , Greer Wenda , Flowerdew Gorden , Bethune Drew , Henteleff Harry , Plourde Madelaine , Mujoomdar Aneil , French Daniel , Johnston Micheal , Marcato Paola , Xu Zhaolin
Published: February 2019

Most lung cancer patients are diagnosed at an advanced stage, limiting their treatment options to chemotherapies that have very low response rate. New therapies that target driver gene mutations (e.g., EGFR, ALK, ROS1, BRAF) are being used to treat patients who have tumors with these mutations. In addition, a type of immunotherapy called immune checkpoint inhibitor is being used to treat lung cancer patients. For instance, patients with tumors that express PD-L1 are responsive to anti-PD-1/PD-L1 therapy. Thus, being able to identify the presence of driver mutations and PD-L1 will help patients to benefit from different therapies. A total of 844 cases of non-small cell lung cancer samples have been profiled for the presence of EGFR, KRAS, BRAF, PIK3CA, and HER2 mutations by SNaPshot/sizing genotyping. Immunohistochemistry (IHC) was used to identify the protein expression of ALK and PD-L1. Histologic examination was performed to determine the pathologic type, grade, and lymphatic/vascular invasion. Statistical analysis revealed a number of correlations between the presence of the mutations, PD-L1 expression and the patient pathologic data. Specifically, it was determined that women had lung tumors with a significantly greater number of EGFR mutations than men (p value = 0.001). Examining the presence of ALK, EGFR, KRAS, BRAF, PIK3CA, and HER2 mutations against the presence of pleural invasions yielded a p-value of 0.071, which implies evidence that different mutations are not associated with pleural invasion. However, EGFR mutations were associated with the absence of vascular and lymphatic invasions in lung cancer patients (p value = 0.001, 0.002 respectively). In addition, while the expression of PD-L1 does not associate with the patients who express KRAS mutation, it is associated with lung cancer patients who express EGFR mutation (p value = 0.002). Knowing the mutational and PD-L1 status in lung cancer patients will help patients benefit from targeted therapies and/or checkpoint inhibitors.

Citation Format: Akram Alwithenani, Marika Forsythe, Mathieu Castonguay, Wenda Greer, Gorden Flowerdew, Drew Bethune, Harry Henteleff, Madelaine Plourde, Aneil Mujoomdar, Daniel French, Micheal Johnston, Paola Marcato, Zhaolin Xu. Investigating targeted driver mutations and PD-L1 expression for improved therapy of non-small cell lung cancer [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr B21.
Abstract B22: Exposure of AZD4547 FGFR-TKI to a lung squamous carcinoma cell line induces resistance by small cell lung cancer-like transformation associated with ASCL1
Authors: Amano Yosuke , Ando Takahiro , Maemura Keita , Sakatini Toshio , Sunohara Mitsuhiro , Watanabe Kosuke , Kage Hidenori
Published: February 2019

Molecular targeted medicine has progressed in non-small cell lung cancer in recent years, and acquired resistance to molecular targeted therapy has become a major problem. Several mechanisms of acquired resistance have been described, such as secondary mutation of targeted genes, bypass pathway activation, and phenotypic transformation, including epithelial-mesenchymal transition and small cell lung cancer transformation. However, detailed mechanisms and the strategy to overcome the resistance have yet to be clarified thoroughly. In this study, we established a squamous lung cancer cell line with acquired resistance to FGFR tyrosine kinase inhibitor in vitro and identified achaete-scute family bHLH transcription factor 1 (ASCL1) overexpression as a mechanism of the resistance to tyrosine kinase inhibitor treatment.

We identified LK-2 as a squamous cell lung cancer cell line that is highly sensitive to FGFR tyrosine kinase inhibitor AZD4547, which is currently evaluated in clinical trial in squamous cell lung cancer, and established its resistant clone in vitro by exposing gradually increasing concentration of AZD4547. The resistant clone was morphologically more round-shaped and expressed ASCL1 significantly higher than the original LK-2 in a microarray analysis, but did not have additional mutations in putative genes responsible for resistance in RNA-sequencing. ASCL1 inhibition with shRNA lentivirus did not restore the sensitivity to AZD4547, but significantly suppressed the growth of the resistant clone compared to the original LK-2. Cyclin A1 (CCNA1) expression was also increased in the resistant clone and was regulated by ASCL1. CCNA1 inhibition also suppressed the growth of the resistant clone.

These results suggest that ASCL1 was one of the key molecules for the acquired resistance to tyrosine kinase inhibitor therapy, and considering the evidence that ASCL1 is a master regulator of small cell lung cancer-like phenotype, it may be related to small cell lung cancer transformation as a mechanism of the resistance. ASCL1 and its downstream molecule CCNA1 may be candidates to overcome the resistance to tyrosine kinase inhibitor therapy.

Citation Format: Yosuke Amano, Takahiro Ando, Keita Maemura, Toshio Sakatini, Mitsuhiro Sunohara, Kosuke Watanabe, Hidenori Kage. Exposure of AZD4547 FGFR-TKI to a lung squamous carcinoma cell line induces resistance by small cell lung cancer-like transformation associated with ASCL1 [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr B22.
Abstract B25: Decoding tumor microenvironment to enhance NSCLC targeted therapy
Authors: Hu Haichuan , Sequist Lecia , Piotrowska Zofia , Mulvey Hillary , Noeen Sundus , Hare Patricia , Kodack David , Hata Aaron , Niederst Matt , Benes Cyril , Engelman Jeff
Published: February 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR04) of the Conference Proceedings.

Citation Format: Haichuan Hu, Lecia Sequist, Zofia Piotrowska, Hillary Mulvey, Sundus Noeen, Patricia Hare, David Kodack, Aaron Hata, Matt Niederst, Cyril Benes, Jeff Engelman. Decoding tumor microenvironment to enhance NSCLC targeted therapy [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr B25.
Abstract IA03: Functional lung cancer genomics through in vivo genome editing
Authors: Winters Ian P. , Rogers Zoë N. , McFarland Christopher D. , Lalgudi Pranav V. , Chiou Shin-Heng , Kay Mark A. , Petrov Dmitri , Winslow Monte M.
Published: February 2019

Cancer genome sequencing has been instrumental in identifying the genomic alterations that occur in human tumors. However, the functional importance of the vast majority of these alterations, both alone and in combination, remains unknown. I will describe methods that integrate tumor barcoding with CRISPR/Cas9-mediated genome editing and ultradeep barcode sequencing to interrogate multiple tumor genotypes simultaneously in autochthonous mouse models of human cancer. We initially used this method to quantify the effects of eleven of the most frequently inactivated genes in human lung adenocarcinoma on tumor growth in vivo. We also investigated the in vivo fitness advantage conferred by inactivation of each of these eleven genes in combination with inactivation of p53 or Lkb1. This map of tumor-suppressive effects for >30 common lung adenocarcinoma genotypes revealed a complex landscape of context dependence and variability in effect strength. I will also describe a platform that integrates multiplexed Cas9-mediated homology-directed repair (HDR) with DNA barcoding and high-throughput sequencing to simultaneously investigate multiple oncogenic alterations in parallel. Using this approach, we introduced a library of nonsynonymous mutations into endogenous Kras in adult somatic cells to initiate tumors. High-throughput sequencing of barcoded Kras HDR alleles from bulk tumor-bearing lung uncovered surprising diversity in Kras variant oncogenicity. These in vivo approaches may redefine our ability to understand how diverse genomic alterations impact tumor initiation, growth, malignant transformation, and therapy responses.

Citation Format: Ian P. Winters, Zoë N. Rogers, Christopher D. McFarland, Pranav V. Lalgudi, Shin-Heng Chiou, Mark A. Kay, Dmitri Petrov, Monte M. Winslow. Functional lung cancer genomics through in vivo genome editing [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA03.
Abstract IA06: Molecular mechanisms of lung cancer development: Between metabolic reprogramming and genomic instability in the field of cancerization
Authors: Rahman Jamshed , Ji Xiangming , Patel Chirayu , Young Jamey , Massion Pierre P.
Published: February 2019

Airway epithelial cells are prone to assault by risk factors and considered to be the primary cell type involved in the field of cancerization. To investigate risk-associated changes in the bronchial epithelium proteome that may offer new insights into molecular pathogenesis of lung cancer, proteins were identified in the airway epithelial cells of bronchial brushings specimens from risk-stratified individuals by shotgun proteomics. Differential expression of selected proteins was validated by parallel reaction monitoring mass spectrometry in an independent set of individual bronchial brushings. We identified over 300 proteins with a significant trend in expression. Pathway analysis revealed enrichment of carbohydrate metabolic enzymes in high-risk individuals. Glucose consumption and lactate production were increased in human bronchial epithelial BEAS2B cells treated with cigarette smoke condensate for seven months. Increased lipid biosynthetic capacity and net reductive carboxylation were revealed by metabolic flux analyses of [U-13C5] glutamine in this in vitro model, suggesting profound metabolic reprogramming in the airway epithelium of high-risk individuals. These results also pointed towards a fundamental role of glutamine and cysteine metabolism in lung cancer progression. Two amino acid transporters, specifically SLC1A5 and SLC7A11, captured our attention for their high level of expression in lung cancer. These transporters are associated with worse survival in lung cancer. A small-molecular inhibitor of SLC1A5 prevents tumor progression in nude mice, and a glutaminase inhibitor CB-839 exhibits radiosensitization to lung cancer cells, an effect that may be mediated by decreased GSH production. Taken together, these studies provide a rationale for new chemopreventive strategies, selection of patients for surveillance programs, and determining how metabolic reprogramming may contribute genomic instability a requirement to cancer development. Further investigation into how glutamine and cystine metabolism contribute to lung cancer development and progression may lead to novel early-interception strategies in lung cancer. This work is supported by NCI CA186145, CA152662, and DoD W81XWH-11-2-0161.

Citation Format: Jamshed Rahman, Xiangming Ji, Chirayu Patel, Jamey Young, Pierre P. Massion. Molecular mechanisms of lung cancer development: Between metabolic reprogramming and genomic instability in the field of cancerization [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA06.
Abstract IA08: Individualized risk based lung cancer screening: The way forward
Authors: Berg Christine D.
Published: February 2019

The United States Preventive Services Task Force (USPSTF) recommends annual low-dose computed tomography (CT) lung-cancer screening, as does the Center for Medicare and Medicaid Services (CMS) and many major medical organizations. The USPSTF criteria, similar to those used for entry into the definitive National Lung Screening Trial, are for persons aged 55-80 years, who currently smoke or quit within the past 15 years, and who have accumulated at least 30 pack-years of cigarette smoking. Risk calculators as compared to micro-simulation modeling as used for the USPSTF can more specifically account for demographic, clinical, and smoking characteristics and personalize risk. The USPSTF criteria may miss smokers at high risk for lung cancer who would have been selected for CT screening by individual risk calculators. Recently, the National Comprehensive Cancer Network guidelines for lung cancer screening added consideration of use of risk models to select ever-smokers for screening. Online risk tools are available to assist individuals and health care providers as to whether lung cancer screening may be helpful.

Several lung cancer risk models are available, and comparisons across them have been done. In this presentation, a comparison we performed of the statistical predictive properties of 8 published risk models in 2 large prospective U.S. cohorts will be presented. Identifying a consensus cost-effective risk threshold to define screening eligibility is crucial. Another issue is how best to account for competing morbidities that might make screening less useful. Perhaps a life-years gained approach could help to adjust for this. Additionally, risk models require accurate risk factor data, which are rarely available in electronic health records and may require special collection. It also poses challenges in the health care delivery setting given its complexity.

To better capture high-risk smokers and prevent premature deaths from lung cancer, eligibility for lung cancer screening should be based on exceeding a cost-effective risk threshold that balances CT screening benefits and harms, using a lung cancer risk tool validated in the U.S. population.

Citation Format: Christine D. Berg. Individualized risk based lung cancer screening: The way forward [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA08.
Abstract IA28: The genomic landscape of SCLC and other neuroendocrine lung tumors
Authors: George Julie
Published: February 2019

Small cell lung cancer (SCLC) accounts for 15% of all lung cancers and belongs to the family of pulmonary neuroendocrine tumors. This tumor subgroup additionally includes large-cell neuroendocrine carcinomas (LCNEC) and pulmonary carcinoids (PCA), which occur in 3% and 2% of all lung cancer patients, respectively. Carcinoids are usually clinically benign, whereas SCLC and LCNEC are characterized as high-grade malignant tumors. Clinical and histologic studies have suggested similarities between SCLC and LCNECs: Both lung tumor subtypes are associated with heavy smoking and reveal an aggressive tumor growth resulting in a poor overall survival.

We have performed comprehensive and multilevel genomic profiling on SCLC and LCNECs, including genome, exome, and transcriptome sequencing. SCLC and LCNEC tumors reveal high mutation rates with an average of 9.5 and 8.6 mutations per megabase, respectively. Our sequencing studies have shown that SCLC tumors are characterized by bi-allelic inactivation of TP53 and RB1 in almost 100% of the cases. LCNECs, on the other hand, are composed of two mutually exclusive subgroups, which we categorized as “type I LCNECs” and “type II LCNECs.” Type I LCNECs harbor TP53 and STK11 or KEAP1 alterations. Despite sharing genomic alterations with lung adenocarcinomas and squamous cell carcinomas, type I LCNECs exhibit a high neuroendocrine profile with close similarity to SCLC tumors. Type II LCNECs harbor loss of TP53 and RB1, thus showing genetic resemblance to SCLC. However, these tumors were revealed to be different from the majority of SCLC with reduced levels of neuroendocrine markers, an upregulated immune signaling profile, and high activity of the NOTCH pathway. Conclusively, our studies show marked biologic differences between high-grade neuroendocrine lung tumors and we emphasize the precise distinction of LCNEC subgroups from SCLC.

Citation Format: Julie George. The genomic landscape of SCLC and other neuroendocrine lung tumors [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA28.
Abstract IA31: Radiation therapy in lung cancer: Recent trends and future directions
Authors: Gomez Daniel R.
Published: February 2019

Radiation techniques for lung cancer have evolved substantially over the past decade. While previously many patients were treated with 3D conformal therapy, with large uniform margins placed to estimate respiratory motion in the “average” tumor, patients are now simulated with sophisticated, real-time monitoring of internal motion such that treatment volumes can be individualized to maximize normal tissue sparing through image-guided radiation therapy (IGRT). In addition, many patients are now treated with intensity-modulated radiation therapy (IMRT), which has served to drastically improve conformality in select cases and increase the proportion of patients in which high doses are achievable. Concurrent with these advances, proton beam therapy (PBT) has been implemented in a limited number of centers, with the hope of utilizing the dosimetric distribution advantage of the Bragg Peak to further reduce toxicity. The role of PBT in both early-stage and locally advanced non-small cell lung cancer continues to be defined, as results from a phase II multicenter randomized trial predominantly in patients with stage II-III disease did not demonstrate superiority with PBT and concurrent chemotherapy when compared to similar regimens with IMRT. Finally, there have been much data produced with regard to dosing regimens. In early-stage disease, there has been increased implementation of stereotactic ablative body radiation (SABR), with prospective data emerging for both peripheral and central lesions. In the locally advanced setting, results from RTOG 0617 demonstrated no difference when comparing 60 Gy vs. 74 Gy in the setting of chemoradiation. There has also been a trend towards hypofractionation for patients who are unable to tolerate concurrent chemoradiation or those with oligometastatic disease, yet the benefit of hypofractionated radiation sequentially vs. combined modality treatment will need to be further assessed in prospective randomized trials, particularly in the era of immunotherapy.

Citation Format: Daniel R. Gomez. Radiation therapy in lung cancer: Recent trends and future directions [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA31.
Abstract IA32: Multiscale modeling of lung cancer
Authors: Gevaert Olivier
Published: February 2019

Vast amounts of biomedical data are now routinely available for lung cancer patients ranging from sequencing of lung tissues to liquid biopsies. In addition, new computational tools for quantitatively analyzing immunohistochemistry (IHC) and radiology images are now available. Multi-scale data is now available for complex diseases at molecular, cellular and tissue scale to establish a more comprehensive view of key biological processes. Intra and inter individual heterogeneities are often quoted as the main challenge for studying cancer. These heterogeneities exist at all scales, from microscopic to macroscopic. However, the technologies used to generate data at a particular scale don’t equally assess the heterogeneity. A single biopsy of a lung tumor does not reflect the potential cellular and spatial heterogeneity. Radiology images on the other hand provide a global picture of the heterogeneity of the complete tumor at the tissue scale.

We propose multi-scale modeling to counter heterogeneity and uncover potentially untapped synergies between different data modalities by integrating information across spatial scales. Multi-scale modeling involves linking information from molecules, cells, tissues, and organs all the way to the organism and the population. We propose to use high dimensional molecular data with cellular and tissue scale image data to develop a statistical multi-scale modeling approach of cancer by tapping the previously understudied information in IHC and radiology images. For example, recent reports showed how to integrate transcriptomic data and CT images resulting in intriguing associations between gene expression signatures and CT image phenotypes. Secondly, multiple studies have shown that EGFR mutation status can be predicted from CT images features, opening up avenues for non-invasive therapy monitoring. In addition, computational analysis of IHC and radiology images is able to predict the overall survival of lung cancer patients. Each of these single reports have shown the potential of multi-scale modeling, however a complete multi-scale model of lung cancer is still under development. Overall, multi-scale modeling can have profound contributions toward predicting diagnosis and treatment by revealing synergies and previously unappreciated relationships. Multi-scale modeling also can contribute to a more fundamental understanding of lung cancer development and can reveal novel insights in how data at different scales are linked to each other.

Citation Format: Olivier Gevaert. Multiscale modeling of lung cancer [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA32.
Abstract IA34: Targeting cellular heterogeneity in lung adenocarcinoma
Authors: Marjanovic Nemanja , Hofree Matan , Canner David , Trakala Marianna , Wu Katherine , Smith Olivia , Kim Jonathan , Amon Angelika , Regev Aviv , Jacks Tyler , Tammela Tuomas
Published: February 2019

Cancer arises from a single cell through a series of acquired mutations, and gradually develops into a complex tissue comprising phenotypically heterogeneous societies of cancer cells. The phenotype, or state, of each tumor cell is influenced by multiple cell-autonomous and cell-extrinsic factors; the diversity of these cellular states poses a challenge for effective cancer therapies. We characterized the evolution of transcriptomes at the single-cell level in genetically engineered murine lung tumors driven by oncogenic Kras. Tumor progression was accompanied by a dramatic increase in cellular heterogeneity, which was further exacerbated upon deletion of the p53 tumor suppressor, allowing tumor progression into advanced adenocarcinomas. This heterogeneity was manifested by the emergence of de-differentiated or transdifferentiated populations, the activation of latent developmental or regenerative programs, as well as heterogeneous activation of signaling pathways. Surprisingly, similar heterogeneous populations emerged in independent tumors irrespective of considerable intertumoral differences in genetic copy number alterations, suggesting a remarkable degree of convergence to achieve a common evolutionary endpoint. We used these data to computationally model cell-state transitions. Finally, treatment of the lung tumors with chemotherapy enabled the identification of cellular states associated with intrinsic sensitivity or resistance. These results associate tumor progression with increasing transcriptional heterogeneity. The identification of unique gene expression programs in cancer cell subpopulations provides a path towards reducing cellular heterogeneity in cancer.

Citation Format: Nemanja Marjanovic, Matan Hofree, David Canner, Marianna Trakala, Katherine Wu, Olivia Smith, Jonathan Kim, Angelika Amon, Aviv Regev, Tyler Jacks, Tuomas Tammela. Targeting cellular heterogeneity in lung adenocarcinoma [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA34.
Abstract PR04: Decoding tumor microenvironment to enhance NSCLC targeted therapy
Authors: Hu Haichuan , Sequist Lecia , Piotrowska Zofia , Mulvey Hillary , Noeen Sundus , Hare Patricia , Kodack David , Hata Aaron , Niederst Matt , Benes Cyril , Engelman Jeff
Published: February 2019

Background: Tyrosine kinase inhibitors (TKI) have yielded promising responses in non-small cell lung cancer (NSCLC) with EGFR mutations and ALK translocations. However, these and other targeted therapies are limited by intrinsic and acquired drug resistance. The previous study from our group investigated tumor autonomous resistance mechanisms by developing patient-derived cancer models (PDCs). In this study, we aimed to decipher the nonautonomous resistance mechanisms via tumor microenvironment by developing patient-derived fibroblast (PDF) models.

Method: Cancer-associated fibroblast cell lines are established directly from individual EGFR mutant NSCLC biopsies. These cell lines, as representative of each patient’s tumor microenvironment, are further subjected to functional analysis. An imaging-based high-throughput platform is developed to screen for nonautonomous resistance by co-culturing PDC and PDF models in vitro. In the parallel, two independent approaches are performed to further identify mechanisms underlying the nonautonomous resistance. These include a drug screen to determine the pathway maintaining the cancer cells’ survival, and a secretomic analysis on the PDFs to identify the plausible cytokine(s) responsible for the resistance.

Result: By co-culturing screening, nonautonomous resistance can be found in a wide spectrum of models. The subsequent drug screen reveals both a canonical HGF-dependent and novel HGF-independent mechanisms contributing to EGFR TKI resistance. Both of these can be explained by the PDF’s variable cytokine secretion and can be overcome by specific therapeutic combinations. Moreover, the microenvironment-driven EGFR TKI resistance has also been validated in vivo. The prevalence of the identified cytokine is further tested in clinical specimens.

Conclusion: PDFs provide a new avenue to explore nonautonomous resistance for targeted therapy. Applying this approach, we identified both the canonical HGF-dependent and novel HGF-independent mechanisms that putatively confer EGFR TKI resistance. Taking EGFR TKI therapy as a paradigm, these findings will be valuable to optimize targeted therapy and to inform the design of personalized pharmaceutical interventions.

This abstract is also being presented as Poster B25.

Citation Format: Haichuan Hu, Lecia Sequist, Zofia Piotrowska, Hillary Mulvey, Sundus Noeen, Patricia Hare, David Kodack, Aaron Hata, Matt Niederst, Cyril Benes, Jeff Engelman. Decoding tumor microenvironment to enhance NSCLC targeted therapy [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr PR04.
Abstract A009: Forty-three week follow-up of a phase 1a clinical trial of a PSA, IL-2, GM-CSF containing prostate cancer therapeutic vaccine in PSA defined biochemical recurrent prostate cancer patients
Authors: Head Jonathan F. , Daniels Gregory A. , McKinney Michelle , Wang-Rodriguez Jessica
Published: February 2019

While progress has occurred in both cellular and immune checkpoint therapy, the benefit of therapeutic cancer vaccines has been difficult to demonstrate. Prostate cancer may be an exception with the modest activity of Sipuleucel T. We have developed a potentially low cost and easy to administer therapeutic vaccine with the combination of tumor antigen (PSA) and biologic adjuvants (IL2 and GM-CSF). This study is a phase 1a clinical trial of a PSA/IL-2/GM-CSF vaccine in biochemically recurrent hormone-naïve and hormone-independent prostate cancer patients. Major inclusion criteria include adenocarcinoma of the prostate, rising serum PSA and no measurable disease. Phase 1a examines the rate of serious adverse events (SAEs) and dose limiting adverse events (DLAEs) in an initial course of 6 vaccinations (“induction vaccination”). Twenty patients enrolled and nineteen patients completed all six intradermal injections (one patient received 5 vaccines, missed week 11 vaccine) of the PSA/IL-2/GM-CSF vaccine at weeks 1, 2, 3, 7, 11 and 15. None of the patients vaccinated in the phase 1a portion had an SAE or DLAE with the most common AE relating to grade 1 injection site reactions. Fifteen of the 18 patients who received vaccines had immune responses to PSA as demonstrated in a lymphocyte blastogenesis assay by week 31. Interestingly, after vaccination 14 of 20 patients had an increase in the doubling time of their serum PSA, suggesting a slowing of the growth of the prostate cancer. Five of 20 patients have progressed (3 PSA progression and 2 radiologic progression) at 43 weeks’ follow-up.

Citation Format: Jonathan F. Head, Gregory A. Daniels, Michelle McKinney, Jessica Wang-Rodriguez. Forty-three week follow-up of a phase 1a clinical trial of a PSA, IL-2, GM-CSF containing prostate cancer therapeutic vaccine in PSA defined biochemical recurrent prostate cancer patients [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A009.
Abstract A030: Identification of α-fetoprotein-specific T-cell receptors for hepatocellular carcinoma immunotherapy
Authors: He Yukai , Zhu Wei , Peng Yibing , Wang Lan , Hong Yuan , Wu Juan , Celis Esteban
Published: February 2019

Hepatocellular carcinoma (HCC) is the major form of liver cancer for which there is no effective therapy. Genetic modification with T-cell receptors (TCR) specific for HCC-associated antigens, such as α-fetoprotein (AFP), can potentially redirect human T-cells to specifically recognize and kill HCC tumor cells to achieve antitumor effects. In this study, by using lentivector and peptide immunization, we identified a population of CD8-T-cells in HLA-A2 transgenic AAD mice that recognized AFP158 epitope on human HCC cells. Adoptive transfer of the AFP158-specific mouse CD8-T-cells eradicated HepG2 tumor xenografts as large as 2cm in diameter in immunocompromised NSG mice. We then established T-cell hybridoma clones from the AFP158-specific mouse CD8-T-cells and identified three sets of paired TCR genes out of 5 hybridomas. Expression of the murine TCR genes redirected primary human T-cells to bind HLA-A2/AFP158 tetramer. The TCR gene-engineered human T-cells (TCR-T) also specifically recognized HLA-A2+AFP+ HepG2 HCC tumor cells and produced effector cytokines. Importantly, the TCR-T-cells could specifically kill HLA-A2+AFP+ HepG2 tumor cells without significant toxicity to normal primary hepatocytes in vitro. Adoptive transfer of the AFP-specific human TCR-T-cells could eradicate HepG2 tumors in NSG mice. Conclusion: We have identified novel AFP-specific murine TCR genes that can redirect human T-cells to specifically recognize and kill HCC tumor cells, and those AFP158-specific TCRs have a great potential to engineer a patient’s autologous T-cells to treat HCC tumors.

Citation Format: Yukai He, Wei Zhu, Yibing Peng, Lan Wang, Yuan Hong, Juan Wu, Esteban Celis. Identification of α-fetoprotein-specific T-cell receptors for hepatocellular carcinoma immunotherapy [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A030.
Abstract A032: Chimeric antigen receptor (CAR) targeted epitope determines optimal CAR spacer length for therapy against medulloblastoma
Authors: Johnson Adam J. , Chang Cindy A. , Baldwin Michael L. , Yokoyama Jason K. , Jensen Michael C.M.
Published: February 2019

Improved therapeutic outcomes for children with embryonal brain tumors (EBTs) hinge on the development and optimization of novel, targeted therapies able to eradicate tumors without serious treatment-related damage to the central nervous system (CNS). To this end, we describe the design and validation of optimized chimeric antigen receptors (CARs) T-cells that target the EBT-associated antigen Her2. Our studies demonstrate that Her2CAR extracellular spacer domains actively influence T-cell in vitro functional outputs and in vivo responses, with the medium (M-) spacer able to confer the greatest antitumor activity against the most common EBT, medulloblastoma. Furthermore, we show that the juxtamembrane epitope targeted on Her2 precluded short spacer Her2CAR activity; yet, this activity could be rescued when the targeted epitope was expressed in a membrane distal position. Similarly, while modifications that abrogate Fc region interactions in the long spacer Her2CAR rescued in vivo activity, the resultant functional outputs failed to reach the antitumor potency elicited by the M-spacer CAR. Here we also demonstrate the in vitro and in vivo activity of M-spacer CAR T-cells produced by our clinical manufacturing process. Results from this preclinical dataset have directed the implementation of a clinical trial that delivers Her2CAR T-cells locoregionally to patients with EBT tumors, coined BrainChild-01. Collectively, these results reiterate the necessity to tailor CARs to their respective targeted antigen epitope and describe the optimization of Her2-targeted CARs for the treatment of EBT tumors.

Citation Format: Adam J. Johnson, Cindy A. Chang, Michael L. Baldwin, Jason K. Yokoyama, Michael C.M. Jensen. Chimeric antigen receptor (CAR) targeted epitope determines optimal CAR spacer length for therapy against medulloblastoma [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A032.
Abstract A034: High-throughput identification of naturally occurring T-cell receptors with therapeutic potential against tumor-associated, viral and neoantigens
Authors: Klinger Mark , Ebert Peter , Osborne Edward , Taniguchi Ruth , Hu Joyce , Hayes Tim , Benzeno Sharon , Carbo Adria , Laur Melanie , Eggers Erica , Robins Harlan
Published: February 2019

The growing use of immunotherapy to treat advanced cancers has brought about a revolution in techniques to mobilize the immune system to antitumor effect. Chimeric antigen receptor (CAR) T-cells targeting CD19 constitute the first modified T-cell products to garner FDA approval for clinical use. However, CAR technologies can only targeT-cell surface antigens, representing approximately one quarter of potential targets. In contrast, T-cell receptor (TCR) therapies can target peptides presented by the major histocompatibility complex (MHC), including those derived from intracellular antigens. Hitherto, such receptors have generally been identified through low-throughput techniques using cancer patients’ blood, followed by affinity-maturation of TCRs, a step that can decrease safety. Here, we demonstrate a novel pipeline for the identification of potential therapeutic TCRs from the naïve repertoire of healthy individuals. Using a technique called Multiplexed Identification of T-cell Receptor Antigen specificity (MIRA), we input hundreds of antigens of interest, including tumor-associated antigens, viral antigens and neoantigens, and identify thousands of TCRs to these antigens. These TCRs then undergo evaluation for affinity, avidity, cytokine release, cytotoxicity and safety. Cytotoxicity is demonstrated using both peptide-loaded and endogenously presented peptides. Safety is evaluated using alanine-glycine scans; evaluation of reactivity of TCRs againsT-cell lines and primary cells is planned. We have fully characterized several TCRs targeting clinically relevant targets, which demonstrated improved avidity and cytolysis relative to a benchmark TCR, and a promising preliminary safety profile. In the recent year, cancer neoantigens as targets for natural and therapeutic antitumor responses have also gained momentum given their attractive product profile. Hereto, we also show data for TCRs against shared neoantigens.

Citation Format: Mark Klinger, Peter Ebert, Edward Osborne, Ruth Taniguchi, Joyce Hu, Tim Hayes, Sharon Benzeno, Adria Carbo, Melanie Laur, Erica Eggers, Harlan Robins. High-throughput identification of naturally occurring T-cell receptors with therapeutic potential against tumor-associated, viral and neoantigens [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A034.