bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 602: Prognostic utility of circulating tumor cells/cell clusters in head and neck cancers
Authors: Zhang Xi , Tang Kai , Hughes Brett , Vasani Sarj , Liu Zhen , Warkiani Majid , Hartel Gunter , Ladwa Rahul , Kenny Liz , Punyadeera Chamindie
Published: July 2021

Background: Metastases remains the major cause of death in head and neck cancer (HNC) patients. Circulating tumour cells (CTCs) that are shed from primary and metastatic sites and circulate in patients' peripheral blood, represent an important window into disease status. Minimally invasive techniques are required to identify at time of diagnosis, which patients will have worse prognosis due to distant relapse.

Method: We collected bloods from treatment naïve, locoregionally advanced HNC patients (n=72) at the time of diagnosis, and CTCs were isolated using a spiral microfluidic device. CTCs were then immobilised onto glass slides and identified by using immunofluorescence staining (Cytokeratin/CD45/DAPI). We then correlated the CTC numbers of each patient to their clinical outcome based on either PET/CT scan and/or clinical assessment after definitive treatment.

Results: CTCs were detected in 39/72 (54%) of patients (ranging from 1-18 CTCs/10 mL blood) at diagnosis. CTC numbers were correlated to TNM staging (p<0.01), especially the Node staging (p<0.01). Patients with ≥2 CTCs at baseline showed significantly worse prognosis than those with no CTCs detected (P<0.001). We also found that baseline CTCs is an independent predictor of the adverse clinical outcome (residual disease, recurrence, and equivocal clinical presentation) even after adjusting to other prognostic factors (hazard ratio=4.613, p<0.05). Patients with CTC cluster had poor prognosis compared to patients with single CTCs.

Conclusion: We were able to predict disease progression based on CTC numbers, thus providing potential additional prognostic information for locally advanced HNC patients. The ability to detect CTCs at diagnosis allows us to stratify the risk in patients as well as to understand the fundamental mechanisms by which CTCs spread. The clinical benefit of establishing the use of CTCs in HNC is likely to translate into better patient selection for treatment intensification and/or de-intensification strategies.

Citation Format: Xi Zhang, Kai Tang, Brett Hughes, Sarj Vasani, Zhen Liu, Majid Warkiani, Gunter Hartel, Rahul Ladwa, Liz Kenny, Chamindie Punyadeera. Prognostic utility of circulating tumor cells/cell clusters in head and neck cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 602.
Abstract 604: PTX-35, a potential first-in-class TNFRSF25 agonist, reduced the suppressive activity of regulatory T cells and enhanced CD4+ T cell effector responses, in the presence of tumor antigens, in a murine melanoma model
Authors: Kalim Khalid W. , Tahiliani Vikas , Miriyala Jayalakshmi , Dillon Patrick J. , Trinh Anh M. , Chaney Abra N. , Seavey Matthew M. , Dixon Eric P. , Jasuja Rahul R. , Hutchins Jeff T.
Published: July 2021

Tumor cells evolve multiple mechanisms to escape immune surveillance, one of which is to establish a tolerogenic microenvironment by recruiting immune suppressive cell types such as regulatory T cells (Tregs). Tumor Necrosis Factor Receptor Super Family 25 (TNFRSF25), also known as Death Receptor 3 (DR3), is a potent costimulatory molecule and is preferentially expressed by activated and antigen-experienced T cells. PTX-35, a potential first-in-class TNFRSF25 agonist antibody, is currently in a Phase 1 clinical trial in patients with solid tumors. The purpose of this study was to characterize the impact of mouse PTX-35 (mPTX-35) on tumor growth as well as Treg stability and functionality, including propensity of Tregs to be converted to effector CD4+ T cell subtypes. In this murine melanoma model, C57BL/6 mice were subcutaneously injected with B16.F10-OVA tumor cells. Mice were administered various combinations of mPTX-35, a PD-1 inhibitor, and tumor antigens. When mPTX-35 was administered with tumor antigens, we observed: (1) reduction in the suppressive activity of Tregs, as reflected by decreased expression of CD25 and Nrp1 among Tregs in the peripheral blood and CTLA-4 in the tumor; (2) compromised Treg stability and increased Treg plasticity, including an increased percentage of RORγt+Foxp3+ unstable Tregs and increased conversion of Tregs to RORγt+Foxp3- Th17 cells; (3) increased effector IFNγ+ Th1 cells and effector IL17+ Th17 cells in blood and tumor microenvironment; and (4) elevation in effector IFNγ+ CD8+ T cells in the blood. Importantly, these changes in Treg stability and functionality were associated with delayed tumor progression. In conclusion, we demonstrate that mPTX-35, in the presence of tumor antigens, resulted in the reprogramming of the Treg cell phenotype, including reduced suppressive activity of Tregs and increased plasticity towards inflammatory Th1 and Th17 cells. PTX-35 has the potential to improve the efficacy of treatment regimens where tumor antigens are released.

Citation Format: Khalid W. Kalim, Vikas Tahiliani, Jayalakshmi Miriyala, Patrick J. Dillon, Anh M. Trinh, Abra N. Chaney, Matthew M. Seavey, Eric P. Dixon, Rahul R. Jasuja, Jeff T. Hutchins. PTX-35, a potential first-in-class TNFRSF25 agonist, reduced the suppressive activity of regulatory T cells and enhanced CD4+ T cell effector responses, in the presence of tumor antigens, in a murine melanoma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 604.
Abstract 610: Paired alterations of 35-gene expressing profiles before and after neoadjuvant chemoradiation in esophageal squamous cell carcinoma patients impact clinical prognosis
Authors: Chen Yu-Jen , Bhan Prerana , Wu Terry , Chang Hang , Lee Chi-Jung
Published: July 2021

Purpose: Neoadjuvant concurrent chemoradiotherapy (CCRT) followed by surgery is widely used for treating locally advanced esophageal cancer in the thorax. The aim of this study was to evaluate alterations of the 35-gene expression profiles in irradiated esophageal tissues before and after chemoradiation.

Materials and Methods: A total number of 50 patients with Esophageal squamous cell carcinoma (ESCC) received neoadjuvant CCRT were retrospectively reviewed. The prescribed radiation dose for gross tumor and metastatic lymph nodes or subclinical mucosal/submucosal disease and regional lymphatic basin were 48 Gy and 43.2 Gy, respectively. All patients underwent CCRT using the intensity-modulated radiation therapy in 24 fractions and received concurrent chemotherapy with cisplatin. A 35-gene expression profile derived from reported ESCC-specific next-generation sequencing was performed. Paired 35-gene expression profiles were analyzed using pre-CCRT biopsy and post-CCRT esophagectomy specimens. In vitro validation of differential expression genes by using human ESCC cell lines 81T and TE2 was performed using the same 35-gene panel.

Results: The alterations in mutation of genes between pre-CCRT biopsy and post-CCRT surgical specimens (delta expression profiles) correlated with pathological complete remission, overall survival and progression free survival. Among the delta expression profiles, the most significant alterations with prognostic impact were noted in MYH4 (p.Gln1210fs) and TTN (p.Asp7145His). Mutation of MYH4 (p.Gln1210fs) and TTN (p.Asp7145His) were demonstrated with favorable and unfavorable impact on rate of pathological complete remission, respectively. The mutation of MYH4 was also noted in human ESCC cell lines 81T and TE2 cells.

Conclusion: By using the 35-gene expression profiles, specific mutations correlated to prognosis in esophageal squamous cell carcinoma patients receiving neoadjuvant chemoradiation and esophagectomy were found. The role of MYH4 mutation in treatment outcome and development as therapeutic target of esophageal squamous cell carcinoma remains to be clarified.

Citation Format: Yu-Jen Chen, Prerana Bhan, Terry Wu, Hang Chang, Chi-Jung Lee. Paired alterations of 35-gene expressing profiles before and after neoadjuvant chemoradiation in esophageal squamous cell carcinoma patients impact clinical prognosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 610.
Abstract 627: Molecular validation of the 8th edition AJCC cancer staging system for resectable pancreatic cancer: Proposal of integrative translational staging system
Authors: Lee Sung Hwan , Lee Sunyoung , Lee Ju-Seog
Published: July 2021

Introduction: Even though the 8th edition AJCC cancer staging system for pancreatic cancer has validated major clinicopathologic factors in multiple clinical cohorts, there is still an unmet need for integrative consideration using multi-omics data to stratify the patients with pancreatic cancer elaborately.

Methods: We performed a comprehensive analysis and profiling using genomic, transcriptomic, and proteomic data from TCGA-PAAD and other translational cohorts (4 cohorts, n=340). Molecular features and major subtypes were analyzed mutually with clinical and pathologic factors, especially the 8th AJCC staging system.

Results: Aggressive molecular subtypes, basal-like and squamous subtype, were significantly associated with a higher nodal stage, but tumor size didn't show a clear association with molecular features. The activated stroma of the pancreatic cancer microenvironment was significantly correlated with poor differentiation and large tumor size. The mutational pattern of KRAS and several transcriptomic pathways, such as epithelial-mesenchymal transition and DNA repair, were differently presented in each clinical stage from the 8th AJCC TNM staging system. The optimal algorithm was identified to show significantly higher performance for the prediction for cancer relapse and cancer-specific survival in discovery and validation cohorts. The in-silico prediction for molecular target agents and immunotherapy was performed for final clusters from the optimal stratification system revealed from the integrative analysis.

Conclusions: Our comprehensive multi-omics analysis reveals clear needs for the combination of clinical staging and molecular profiling and provides crucial evidence for precision strategy in patients with resectable pancreatic cancer.

Citation Format: Sung Hwan Lee, Sunyoung Lee, Ju-Seog Lee. Molecular validation of the 8th edition AJCC cancer staging system for resectable pancreatic cancer: Proposal of integrative translational staging system [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 627.
Abstract 632: Growth hormone deficiency in childhood germ cell tumor survivors
Authors: Lone Diana W. , Sadak Karim T. , Miller Bradley S. , Hubbard Aubrey K. , Sample Jeannette , Roesler Michelle , Poynter Jenny N.
Published: July 2021

Introduction: Germ cell tumors (GCTs) are a rare heterogenous group of cancers accounting for 3% of childhood cancers and 15% of cancers in adolescents. Cure rates for GCTs exceed 90% and GCTs currently comprise the 3rd largest group of childhood cancer survivors in the United States. Little systematic evidence exists on late effects of cancer treatment in this group. Treatment for GCTs includes some combination of chemotherapy, radiation, and gonadectomy; interventions that place survivors at high risk for developing endocrinopathies. One of the most common endocrinopathies experienced by childhood cancer survivors is growth hormone deficiency (GHD). In this analysis, we evaluated the prevalence of growth hormone deficiency among GCT survivors.

Methods: Participants were previously enrolled in the Germ Cell Tumor Epidemiology Study (GaMETES), which is a case parent triad study conducted using the Children's Oncology Group registry protocols. Data on late effects and outcomes are currently available for a subset of the participants who have provided additional consent for a follow-up study including a self-administered questionnaire and medical record retrieval. GHD was identified via self-report and validated through medical records. Treatment information was abstracted from the medical records.

Results: In this interim analysis, 230 participants completed the self-administered questionnaire, including 61 with ovarian tumors, 40 with testicular tumors, 38 extragonadal tumors, and 92 intracranial tumors. Thirty-six of the participants reported having growth hormone deficiency (GHD), all of whom had intracranial germ cell tumors (iGCTs). Thus, the prevalence of GHD among iGCT survivors was 39.1%. We validated 45 questionnaire responses against chart review of iGCT patients. Among these 45, 24 (53.33%) iGCT survivors had GHD. Participants who were treated with high doses of cranial radiation (>30 Gy) were more likely to have GHD (OR = 2.3, 95% CI 0.4-13.1).

Conclusions: Growth hormone deficiency is highly prevalent in survivors of childhood intracranial germ cell tumors. The increased prevalence is likely related to the propensity for germ cell tumor involvement of the pituitary stalk, chemotherapy and cranial radiation. Exposure to higher cumulative doses of cranial radiation is associated with higher risk for developing growth hormone deficiency.

Citation Format: Diana W. Lone, Karim T. Sadak, Bradley S. Miller, Aubrey K. Hubbard, Jeannette Sample, Michelle Roesler, Jenny N. Poynter. Growth hormone deficiency in childhood germ cell tumor survivors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 632.
Abstract 635: Response to novel imipridone combination therapies targeting H3K27M mutant diffuse midline glioma (DMG)
Authors: Borsuk Robyn , Zhou Lanlan , Zhang Yiqun , Prabhu Varun V. , Allen Joshua E. , Tapinos Nikos , Lulla Rishi , El-Deiry Wafik S.
Published: July 2021

ONC201, a first-in-class, small molecule, imipridone therapy, is known to induce apoptosis via upregulation of TNF-related apoptosis inducing ligand (TRAIL) and its proapoptotic death receptor (DR5), independent of p53. Current trials have been exploring ONC201 as a monotherapy or in conjunction with radiotherapy, in treatment of patients with newly diagnosed and refractory diffuse intrinsic pontine gliomas (DIPG), a disease which remains incurable at present. We sought to identify synergy between ONC201 or second generation impridones (ONC206 and ONC212), and other chemotherapeutics with known promising pre-clinical activity against DMGs.

Seven patient derived H3K27M mutant DIPG cell lines (SU-DIPG-IV, SU-DIPG-13, SU-DIPG-25, SUDIPG-27, SU-DIPG-29, SU-DIPG-36, SF8628) were grown in culture and exposed to ONC201, ONC206, or ONC212 either as monotherapy or in combination with other FDA approved chemotherapeutics (histone de-acetylase inhibitors [HDACi], marizomib, etoposide, and temozolomide). Dose dependent response to imipridone therapy was noted in all cell lines, with mean half maximal inhibitory concentration (IC50) of 1.46 µM in ONC201, 0.11 µM in ONC206, and 0.03 µM in ONC212. Synergy was identified via combination studies, with combination indices of <1 indicating some degree of synergy, and <0.5 indicating very strong synergy. We identified strong synergy between ONC201 and HDAC inhibitors: Panobinostat (CI 0.01) and Romidepsin (CI 0.08), with lesser synergy detected with Entinostat (CI 0.59). Romidepsin similarly showed robust synergy with ONC206 (CI 0.1), as did proteasome inhibitor Marizomib (CI 0.1). Combination of ONC212 and Panobinostat demonstrated induction of apoptosis as measured by the induction of poly (ADP-ribose) polymerase (PARP) cleavage. Our results suggest an increased sensitivity of H3K27M DIPG cell lines to second generation imipridones as compared to ONC201. Additionally, we have shown promising synergism between imipridones with Panobinostat or Romidepsin, which should be considered for clinical translation.

Citation Format: Robyn Borsuk, Lanlan Zhou, Yiqun Zhang, Varun V. Prabhu, Joshua E. Allen, Nikos Tapinos, Rishi Lulla, Wafik S. El-Deiry. Response to novel imipridone combination therapies targeting H3K27M mutant diffuse midline glioma (DMG) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 635.
Abstract 638: Enhancing the molecular diagnosis, prognosis, and treatment monitoring of retinoblastoma using cell free DNA in aqueous humor
Authors: Xu Liya , Kim Mary E. , Polski Ashley , Prabakar Rishvanth K. , Peng Chen-Ching , Chévez-Barrios Patricia , Kim Jonathan W. , Shah Rachana , Jubran Rima , Kuhn Peter , Cobrinik David , Hicks James , Berry Jesse L.
Published: July 2021

Purpose: All previous studies of retinoblastoma (RB) aqueous humor (AH) analysis for tumor-derived cell-free DNA (cfDNA) have focused on sampling AH from eyes undergoing therapy; results from AH liquid biopsy at the time of diagnosis have not yet been published. Herein, we detail the diagnostic and prognostic genomic biomarkers found in the AH cfDNA from 7 RB eyes at diagnosis, each with longitudinal evaluation of over 12 months in follow-up.

Methods: Subjects included 7 eyes of 6 RB patients who underwent AH sampling at diagnosis. CfDNA from each AH sample was isolated and sequenced to assess genome-wide somatic copy number alterations (SCNAs), followed by targeted resequencing and mutation detection using a custom hybridization panel for RB1 and MYCN. Results were compared to peripheral blood RB1 testing and matched tumor samples, when available. Tumor fraction (TFx) was calculated using ichorCNA.

Results: Five of 7 diagnostic AH samples contained highly recurrent RB SCNAs, 4 with large scale alterations and 1 with a focal RB1 gene deletion. Mutational analysis of AH cfDNA identified pathogenic somatic variants in 5 diagnostic AH samples with high variant allele frequency, while the remaining 2 diagnostic AH samples had either a high TFx from SCNAs or a focal MYCN amplification. Taken together, somatic tumoral genomic information was detected in all 7 diagnostic AH samples. The 2 eyes that required enucleation had poor prognostic biomarkers (chromosome 6p gain and MYCN amplification) present at the time of diagnosis. TFx from longitudinal AH sampling corresponded to treatment response over time in all cases for which sequential AH samples were available.

Conclusions: This study demonstrates that AH sampling at diagnosis is both feasible and safe. Molecular profiling of AH provides a plethora of diagnostic and prognostic information from only a single 0.1 mL AH sample.

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Citation Format: Liya Xu, Mary E. Kim, Ashley Polski, Rishvanth K. Prabakar, Chen-Ching Peng, Patricia Chévez-Barrios, Jonathan W. Kim, Rachana Shah, Rima Jubran, Peter Kuhn, David Cobrinik, James Hicks, Jesse L. Berry. Enhancing the molecular diagnosis, prognosis, and treatment monitoring of retinoblastoma using cell free DNA in aqueous humor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 638.
Abstract 645: Network-based assessment of HDAC6 activity is highly predictive of pre-clinical and clinical responses to the HDAC6 inhibitor ricolinostat
Authors: Pan Qingfei , Zeleke Tizita , Chiuzan Cody , Onishi Maika , Alvarez Mariano , Honan Erin , Yang Min , Chia Pei Ling , Mukhopadhyay Partha , Kelly Sean , Wu Ruby , Fenn Kathleen , Trivedi Meghna , Accordino Melissa , Crew Katherine , Hershman Dawn , Maurer Matthew , Jones Simon , Califano Andrea , Kalinsky Kevin , Yu Jiyang , Silva Jose
Published: July 2021

Despite the anticancer activity of pan-histone deacetylase (HDAC) inhibitors, their clinical use has been limited due to toxicity. However, the development of more specific inhibitors that selectively inhibit individual HDACs is emerging as a novel and well-tolerated alternative. Here, we present the results of the first clinical trial evaluating the activity of ricolinostat (the leading HDAC6 inhibitor) in breast cancer (BC) patients. We have developed a computational network-based algorithm to evaluate the activity of the HDAC6 protein, based on the enrichment of its transcriptional targets in differentially expressed genes (HDAC6 score). Through preclinical in vitro and in vivo studies, we confirmed that the HDAC6 score can stratify the sensitivity of BC cells to ricolinostat treatment and may thus have value as a predictive biomarker. Moreover, analysis of ~3,000 primary human breast cancers showed that ~30% of them present high HDAC6 scores. Based on these results, we designed a phase Ib clinical trial to evaluate the activity of ricolinostat plus nab-paclitaxel in metastatic BC patients. Study results showed that the two agents can be safely combined, that clinical activity is identified specifically in patients with HR+/HER2- disease, and that the HDAC6 score was predictive of response. Expansion of our analysis to other tumor types identified multiple cohorts enriched in high HDAC6 score samples. These results suggest that the HDAC6 score may provide an effective, CLIA certified predictive biomarker of ricolinostat sensitivity in multiple human cancers.

Citation Format: Qingfei Pan, Tizita Zeleke, Cody Chiuzan, Maika Onishi, Mariano Alvarez, Erin Honan, Min Yang, Pei Ling Chia, Partha Mukhopadhyay, Sean Kelly, Ruby Wu, Kathleen Fenn, Meghna Trivedi, Melissa Accordino, Katherine Crew, Dawn Hershman, Matthew Maurer, Simon Jones, Andrea Califano, Kevin Kalinsky, Jiyang Yu, Jose Silva. Network-based assessment of HDAC6 activity is highly predictive of pre-clinical and clinical responses to the HDAC6 inhibitor ricolinostat [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 645.
Abstract 663: Longitudinal AXL assessment of circulating tumor cells (CTCs) and its clinical implication in the patients with EGFR mutated non-small cell lung cancer (NSCLC)
Authors: Choi Mi Young , Moon Da Hye , Lee Hae Ung , Jo Jong Min , Sohn Young Woong , Jeon Byung Hee , Kim Sang-We
Published: July 2021

Gefitinib for EGFR mutated non-small cell lung cancer (NSCLC) is showing acquired resistance after 8-11 months, and about 60% of its mechanisms have been reported due to T790M mutation. In about a third of NSCLC, AXL expression can be observed at baseline status. And because AXL expression could be a mechanism of drug resistance, we decided to monitor AXL expression before and after Gefitinib treatment in EGFR mutated NSCLC patients through circulating tumor cells (CTCs). Herein, Thirty-seven out of 55 participants who showed acquired resistance to EGFR-TKI were selected in this prospective analysis. To evaluate the AXL expression on the CTCs, serial blood was obtained (tracking for 4, 12, 24 and 36 weeks after EGFR-TKIs). Baseline tumor tissues were also obtained for genetic analysis. Progression-free survival (PFS) and overall survival (OS) were explored according to the baseline AXL expression. AXL expression was analyzed by observation and quantitative intensity-dependent fold change analysis using immunofluorescence analysis. CTCs were isolated and analyzed by CytoGen's Smart Liquid Biopsy platform that uses the size-based gravity filtration for CTC isolation. We found out that there were ten cases (27%) of the increasing pattern of AXL expression in CTCs during longitudinal monitoring in patients with progressive disease (PD). The most remarkable aspect is that AXL up-regulation in CTCs preceded the time point of clinical PD. These results imply that the expectation of clinical PD by blood CTC (liquid biopsy-based examination) can be combined with the T790M. Moreover, the PFS (p= 0.002) and OS (p= 0.007) were both worse in patients with AXL expression at the baseline status compared with patients without AXL expression (N=55). Conclusively, this study implicates that AXL in CTCs could be deployed for prognostic evaluation. And the clinical monitoring of specific target marker in CTC could help for further treatment strategy.

Citation Format: Mi Young Choi, Da Hye Moon, Hae Ung Lee, Jong Min Jo, Young Woong Sohn, Byung Hee Jeon, Sang-We Kim. Longitudinal AXL assessment of circulating tumor cells (CTCs) and its clinical implication in the patients with EGFR mutated non-small cell lung cancer (NSCLC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 663.
Abstract 665: Cannabinoid receptor 2 tumor tissue gene expression positively affects lung cancer survival following radical surgery
Authors: Srovnal Josef , Vidlarova Monika , Berta Emil , Prasil Petr , Kourilova Pavla , Chudacek Josef , Hajduch Marian
Published: July 2021

Background: Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related death with a 5-year survival rate of 21%. New treatment options are thus required, and the endogenous cannabinoid system is a potential new therapeutic target. We therefore analysed the expression of cannabinoid 1 and 2 receptor (CB1 and CB2) genes in NSCLC tissue and its influence on survival following radical surgery.

Method: CB1 and CB2 gene expression in tumor tissue samples collected from 100 NSCLC patients was analysed using the quantitative real-time polymerase chain reaction. Since gene expression may not correlate with protein production, the presence of the CB2 protein was validated by immunohistochemistry. CB2 gene expression was correlated with disease stage, the presence of lymph node and distant metastases, tumor size, tumor grade, and survival.

Result: Patients with tumors expressing CB2 gene had significantly longer overall survival (OS) (p<0.001), cancer-specific survival (CSS) (p=0.002), and disease-free survival (DFS) (p<0.001). They also had less advanced disease stage (p=0.047) and fewer lymph node metastases (p=0.011) at the time of surgery. A multivariate analysis showed that CB2 tumor tissue gene expression was a positive prognostic factor for OS (Hazard ratio (HR) = 0.402, p=0.013), CSS (HR=0.344, p=0.025), and DFS (HR=0.274, p=0.003). CB1 gene expression had no effect on survival. CB1 and CB2 gene expression was detected in 50% and 79.6% of the tumor tissue samples, respectively, and immunohistochemistry confirmed a correlation between CB2 gene expression and the presence of the CB2 protein in tumor tissues.

Conclusion: In NSCLC patients undergoing radical surgery, CB2 gene expression in tumor tissue is associated with less advanced disease stage, fewer lymph node metastases, and longer survival (OS, CSS, DFS).

Acknowledgement: This study was supported by Ministry of Health of the Czech Republic (NV18-03-00470), Ministry of Education, Youth and Sport of the Czech Republic (LM2018132), Palacky University Olomouc (LF 2020_007), Technological Agency of the Czech Republic (TN01000013), European Regional Development Fund (ENOCH CZ.02.1.01/0.0/0.0/16_019/0000868) and Cancer Research Czech Republic.

Citation Format: Josef Srovnal, Monika Vidlarova, Emil Berta, Petr Prasil, Pavla Kourilova, Josef Chudacek, Marian Hajduch. Cannabinoid receptor 2 tumor tissue gene expression positively affects lung cancer survival following radical surgery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 665.
Abstract 67: Paralleled convergence of antigen load, disease progression and systemic T cell exhaustion in lung cancer and HIV-1 infection
Authors: Reading James L. , Rosenthal Rachel , Black Lucas , Ung Seng Kuong , Streatfield Claire , Wooding Dominic , Esbjornsson Joakim , Hunter Eric , Quezada Sergio A. , Hare Jonathan , Swanton Charles
Published: July 2021

Fatal immune failure in untreated HIV-1 infection is co-defined by evolution of viral escape mutants, increasing antigen diversity and concomitant T cell exhaustion. We have recently described mutational heterogeneity as a negative prognostic factor in primary non-small cell lung cancer (NSCLC), highlighted pathways of neoantigen-driven immune escape and reported a program of TMB-associated T cell exhaustion. Here we present interim analysis from a direct comparison of antigen evolution, peripheral T cell exhaustion and disease progression in 279 longitudinal samples from 64 HIV-1 infected volunteers in the retrospective IAVI protocol C cohort and 85 longitudinal samples from 35 NSCLC patients in the TRACERx study. Systemic PD1hi TOX+TCF7- Tex cell subsets were associated with virus load and CD4 loss in HIV-1 infection and tumour burden and disease progression in NSCLC. Levels of systemic Tex cells in NSCLC tracked with the extent of compartment-wide intra-tumoral CD4 and CD8 T cell dysfunction. Tex populations contracted upon antigen loss via ART in HIV-1, a process paralleled during surgical resection in NSCLC. Resurrection of systemic Tex populations was observed at disease recurrence in NSCLC, mediated in-part by post-operative persistence and re-expansion of minor circulating Tex subsets. These data highlight a common convergence of T cell exhaustion, antigen load and disease progression in distinct clinical conditions of impaired immune surveillance and chronic antigen stimulation.

Citation Format: James L. Reading, Rachel Rosenthal, Lucas Black, Seng Kuong Ung, Claire Streatfield, Dominic Wooding, Joakim Esbjornsson, Eric Hunter, Sergio A. Quezada, Jonathan Hare, Charles Swanton. Paralleled convergence of antigen load, disease progression and systemic T cell exhaustion in lung cancer and HIV-1 infection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 67.
Abstract 689: Strong anti-tumor activity and stability of IL-2/anti IL-2 conjugate SLC-3010 in preclinical experiments
Authors: Kim Geon-A , Kim Da-Eun , Kim Yoo-Jin , Han Kyung-Mi , Kim Hee-Sung , Park Soon-Hye , Park Jae-Hyun , Roh Eun-Jihn , Wie Gil-Tae , Lee Jun-Young
Published: July 2021

To achieve successful anti-cancer response with IL-2, it is necessary to develop a method delivering selective IL-2 signaling on CD8 T and NK cells than Tregs We report preclinical results with SLC-3010 that consist of IL-2 conjugate with anti-IL-2 antibody named TCB2 TCB2 is a humanized mouse antibody that specifically recognize the epitope on IL-2 where interact with CD25 as demonstrated with crystallography To develop SLC-3010, stable cell lines producing wild type human IL-2 or TCB2 were separately manufactured using mammalian cell system The pharmacokinetics analysis of SLC-3010 revealed 8-12h of half-life in mouse Although SLC-3010 dose not employ any artificial linker between IL-2 and TCB2, the half-life of SLC-3010 is comparable with other IL-2 based molecules thereby demonstrating stable binding of TCB2 against IL-2 under physiological condition In mouse model, single dosing of SLC-3010 preferentially stimulates CD8 T cells than Tregs in both secondary lymphoid organ and tumor microenvironment SLC-3010 exhibited strong anti-tumor activity which is demonstrated with circulating B cell lymphoma, metastatic solid tumor, and traditional solid tumor models Furthermore, synergistic anti-tumor response was observed by SLC-3010 with various anti-cancer drugs including chemotherapeutic reagents, targeted therapeutics, and checkpoint inhibitors such as anti PD-1 and anti CTLA-4 Once the transplanted tumor was completely cured by SLC-3010, the mice acquired strong memory response against the rejected tumor indicating strong immunological memory formation against recurrent tumors Non-human primate (NHP) efficacy test revealed that single dosing of SLC-3010 expanded CD8 T and NK cells in dose dependent manner, hence the number of CD8 T cells were increased up to 5 fold in the PBMC population with non-toxic dose of SLC3010 The peak of response in NHP was day 6-7 as analyzed the number of CD8 T cells in PBMC The strong activation of CD8 and NK cells with NHP model indicating that the binding of TCB2 and IL-2 is stable in the system as well The SLC-3010 induced selective stimulation of CD8 T cells was also demonstrated with human T cells using humanized NSG mice thereby providing an insight for developing future clinical trials with SLC-3010.

Citation Format: Geon-A Kim, Da-Eun Kim, Yoo-Jin Kim, Kyung-Mi Han, Hee-Sung Kim, Soon-Hye Park, Jae-Hyun Park, Eun-Jihn Roh, Gil-Tae Wie, Jun-Young Lee. Strong anti-tumor activity and stability of IL-2/anti IL-2 conjugate SLC-3010 in preclinical experiments [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 689.
Abstract 71: The immunocytokine PD1-IL2v overcomes immune checkpoint resistance, and combination with an anti-PD-L1 antibody further enhances its anti-tumor activity
Authors: Wullschleger Stephan , Tichet Mélanie , Codarri-Deak Laura , Umana Pablo , Klein Christian , Hanahan Douglas
Published: July 2021

Cancer immunotherapies have shown therapeutic benefits in the clinic, but treatments involving immuno-stimulatory agents such as interleukin 2 (IL-2) are typically accompanied by severe adverse events. One strategy to limit the systemic toxicity of IL-2 is to target it specifically to the tumor microenvironment. Here we evaluated the therapeutic efficacy of the bi-specific immunocytokine PD1-IL2v in the genetically engineered, spontaneous RIP1-Tag5 (RT5) mouse model of pancreatic neuroendocrine tumors (PanNETs). PD1-IL2v is a bi-specific molecule based on a bivalent PD-1 antibody to which a single IL2 variant (IL2v) is fused. PD-1 serves as a targeting moiety to deliver IL2v in cis to PD-1-positive T cells located in the immune tumor microenvironment, and IL2v has been engineered to lack binding to CD25 in order to selectively expand effector T cells but not immuno-suppressive regulatory T cells (Tregs). Although RT5 mice are capable of mounting an adaptive immune response against the tumor-driving oncoprotein, PanNETs developing in RT5 are resistant to immune checkpoint inhibitors. In comparing the untargeted version of IL2v combined with an anti-PD-1 antibody vs. the PD-1 targeted immunocytokine PD1-IL2v, we found that PD1-IL2v, but not the mixture, produced significant anti-tumor activity. PD1-IL2v treatment resulted in a substantial infiltration of CD8+ T cells into the pancreatic islet tumors, which in the context of sustained treatment with PD1-IL2v resulted in tumor regression. Interestingly, following tumor shrinkage, the PanNETs appeared to stabilize and eventually relapsed. Notably, we identified up-regulation of PD-L1 on the tumor vasculature in relapsing tumors as a potential factor in the observed adaptive resistance, which encouraged combining an anti-PD-L1 antibody with PD1-IL2v, leading to enhanced therapeutic benefit. The data obtained in this spontaneous de novo tumor model motivate consideration of evaluating the PD1-IL2v/anti-PD-L1 combination therapy in the clinical setting, in particular in anti-PD-1/anti-PD-L1 resistant tumors infiltrated with PD1+CD8+ T cells.

Citation Format: Stephan Wullschleger, Mélanie Tichet, Laura Codarri-Deak, Pablo Umana, Christian Klein, Douglas Hanahan. The immunocytokine PD1-IL2v overcomes immune checkpoint resistance, and combination with an anti-PD-L1 antibody further enhances its anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 71.
Abstract 724: FE65 in breast cancer patients and its clinicopathological significance
Authors: Xu Junyao , Iwabuchi Erina , Miki Yasuhiro , Kanai Ayako , Ishida Takanori , Sasano Hironobu
Published: July 2021

FE65 is well known to bind with amyloid precursor protein (APP), translocate FE65-APP complex into nucleus and modulate gene transcription. APP was reported to be closely associated with breast cancer progression but clinical significance of FE65 has remained virtually unknow. We first studied 88 surgical pathology specimens with their clinicopathological factors. Results showed nuclear FE65 immunoreactivity was significant positively correlated with estrogen receptor (ER) status (P=0.054) and inversely with histological grade of the patients(P=0.038). We then analyzed 145 ER positive cases which include 44 ductal carcinomas in situ (DCIS) cases and 101 invasive ductal carcinoma (IDC) cases. The number of FE65 positive carcinoma cells was significantly higher in IDC than DCIS (P=0.0395). We then performed in vitro wound healing assay, which demonstrated that FE65 knock-down cells migrated slower than control groups (P=0.0495). FE65 formed the complex with APP and in order to further elucidate their detailed intracellular binding status, proximity ligation assay (PLA) was employed. Binding of APP and FE65 was detected in the cytoplasm with that of ER and FE65 in the nucleus. These results demonstrated that FE65 first bound to APP in the cytoplasm and was then released into the nucleus after phosphorylation of APP. This released FE65 in the nuclei could interact with ER to promote cell migration. Based on those results above, we then conducted the WST-8 assay to further elucidate the possible involvement of FE65 in cell survival. FE65 influenced response of tamoxifen in breast carcinoma cells, and the survival rate of those knocked down by FE65 siRNA was significantly higher than that of the control siRNA group(P=0.0039), indicating that FE65 could contribute to increase the sensitivity of tamoxifen action.In summary, FE65 was demonstrated to stimulate cell proliferation and migration, possibly as a result of the interaction of FE65 with APP in the cytoplasm and ER in the nuclei. In addition, FE65 could enhance the actions of tamoxifen by increasing ER expression and subsequent availability of its binding to ER.

Citation Format: Junyao Xu, Erina Iwabuchi, Yasuhiro Miki, Ayako Kanai, Takanori Ishida, Hironobu Sasano. FE65 in breast cancer patients and its clinicopathological significance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 724.
Abstract 725: LIFR promotes SUCLG2-dependent castration resistance and neuroendocrine differentiation of prostate cancer
Authors: Yeh Hsiu-Lien , Lin Shian-Ren , Jiang Kuo-Ching , Chen Wei-Hao , Liu Yen-Nien
Published: July 2021

Neuroendocrine (NE) differentiation is a well-recognized phenotypic change of prostate cancer after androgen-deprivation therapy (ADT) that ultimately develops an aggressive subtype. An increasing number of patients with NE histologic type prostate cancer present with metabolic genes disorder. However, the metabolic pathways that influence the malignant transformation and the NE differentiation of prostate cancer remain unclear. In this study, we found that ADT induced the upregulation of leukemia inhibitory factor receptor (LIFR), which contributes to the expression of succinate-CoA ligase GDP-forming beta subunit (SUCLG2), thus activating metabolic reprogramming to enable NE differentiation and increase aggressiveness. Activated LIF-LIFR-STAT3 axis stimulates SUCLG2 expression, subsequently upregulating glucose metabolism and the NE differentiation of prostate cancer. SUCLG2 knockdown markedly reduced mitochondrial DNA content and inhibited the enzymatic activity of succinyl-CoA synthetase and nucleoside diphosphate kinase (NDPK) in prostate cancer cells. SUCLG2 overexpression promoted androgen-independent proliferation; however, SUCLG2 knockdown suppressed NE differentiation and the ADT resistance of prostate cancer cells. Analysis of prostate tissue samples showed an increased intensity of LIFR associated with SUCLG2 in castration-resistant prostate cancer tumors. Our study identifies SUCLG2 as a potential prognostic marker of ADT-resistant prostate cancer and suggests it could be a target for prostate cancer treatment.

Citation Format: Hsiu-Lien Yeh, Shian-Ren Lin, Kuo-Ching Jiang, Wei-Hao Chen, Yen-Nien Liu. LIFR promotes SUCLG2-dependent castration resistance and neuroendocrine differentiation of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 725.
Abstract 726: Correlation between T cell infiltration subtypes and intratumoral hormone levels in endometrial cancer
Authors: Miki Yasuhiro , Onuma Fuka , Takagi Kiyoshi , Iwabuchi Erina , Suzuki Takashi , Sasano Hironobu , Ito Kiyoshi
Published: July 2021

T cells in the tumor microenvironment, or tumor-infiltrating lymphocytes (TILs), have been recognized to play a critical role in tumor immunity. TILs are heterogeneously defined, and different T cell subsets have various effects. The properties of some T cell markers, such as CD3, CD4, CD8, and forkhead box P3 (FOXP3), are poor prognostic factors in several types of cancers. The effects of sex steroid hormones on T cell function in inflammation are known, and estrogen and androgen regulate the secretion of cytokines to enhance and attenuate, respectively, the immune response. While intratumoral estrogen and androgen synthesis plays an important role in the progression of cancer cells in hormone-dependent cancers, the relationship between intratumoral hormone levels and TIL subtypes is unclear. In this study, we examined the correlation between intratumoral hormone concentrations and expression scores of TIL markers in endometrial cancer. We employed frozen and formalin-fixed paraffin-embedded (FFPE) tissues of 19 endometrial cancer tissues (G1, n=6; G2, n=5; G3, n=6; Serous, n=2). Hormone concentrations were measured by LC-MC/MS from the frozen sample, and immunohistochemistry of T cell markers was performed using FFPE tissues. The five T cell markers examined in this study were CD3 (total T cells), CD4 (helper T cells), CD8 (cytotoxic T cells), FOXP3 (regulatory T cells), and thymocyte selection-associated HMG box (TOX) (exhausted T cells). The number of intratumoral CD3 in an arbitrary two-field (magnification, ×200) was counted and used as the CD3 score. Similarly, the number of other markers was counted, and the ratio to CD3 was used as the score for each marker. A correlation was found between TOX and CD8 using correlation between markers. The median scores for CD3 and CD8 were significantly higher in type 1 (G1 and G2) than in type 2 (G3 and serous) (CD3, p=0.031; CD8, P=0.031). The CD4 score had a positive correlation with estrogen (estrone, r2=0.591; estradiol, r2=0.501) and androgen (dihydrotestosterone, r2=0.327) concentrations. There was a poor negative correlation between cortisol concentration and CD8 score (r2=0.203). Estrogen and androgen at the tumor in situ were considered to affect TILs, especially CD4 positive T cells. It is also suggested that cortisol acts on immunosuppression, even locally in the tumor. This study showed the potential of a combination of immunotherapy and hormone therapy in endometrial cancer.

Citation Format: Yasuhiro Miki, Fuka Onuma, Kiyoshi Takagi, Erina Iwabuchi, Takashi Suzuki, Hironobu Sasano, Kiyoshi Ito. Correlation between T cell infiltration subtypes and intratumoral hormone levels in endometrial cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 726.
Abstract 729: Prognostic significance of COX-2 over-expression in BRAF mutated Middle Eastern PTC
Authors: Bu Rong , Parvathareddy Sandeep K. , Siraj Abdul K. , Annaiyappanaidu Padmanaban , Iqbal Kaleem , Rasheed Maha Al , Haqawi Wael , Al-Kuraya Khawla S.
Published: July 2021

The cyclooxygenase-2 (COX-2) - prostaglandin E2 (PGE2) pathway has been implicated in carcinogenesis with BRAF mutation shown to promote PGE2 synthesis. This study was conducted to evaluate COX-2 expression in a large cohort of Middle Eastern Papillary Thyroid Carcinoma (PTC) and further evaluate the prognostic significance of COX-2 expression in strata of BRAF mutation status. BRAF mutation analysis was performed by Sanger sequencing and COX-2 expression was evaluated immunohistochemically using tissue microarray. COX-2 over-expression was noted in 43.2% (567/1314) of cases and was significantly associated with poor prognostic markers such as extra-thyroidal extension, lymph node metastasis and higher tumor stage. COX-2 was also an independent predictor of poor disease-free survival (DFS). Most notably, the association of COX-2 expression with DFS differed by BRAF mutation status. COX-2 over-expression was associated with poor DFS in BRAF mutant but not BRAF wild-type PTCs with the multivariate-adjusted hazard ratio of 2.10 (95% CI = 1.52 - 2.92; p < 0.0001) for COX-2 over-expressed tumors in BRAF mutant PTC. In conclusion, the current study shows that COX-2 plays a key role in prognosis of PTC patients. Furthermore, a significant association of COX-2 expression with poor DFS was noted in BRAF mutated tumors. Our data suggests the potential therapeutic role of COX-2 inhibition in patients with BRAF mutated PTC.

Citation Format: Rong Bu, Sandeep K. Parvathareddy, Abdul K. Siraj, Padmanaban Annaiyappanaidu, Kaleem Iqbal, Maha Al Rasheed, Wael Haqawi, Khawla S. Al-Kuraya. Prognostic significance of COX-2 over-expression in BRAF mutated Middle Eastern PTC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 729.