bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5895: Examining the association between arsenic exposure and cancer risk among Canadian adults
Authors: Pullella Katherine , Chang Vicky C. , Harris Shelley A. , Hanley Anthony J. , McLaughlin John R. , Lubinski Jan , Narod Steven A. , Kotsopoulos Joanne
Published: June 2022

Background: Arsenic is a highly prevalent environmental toxin and an established carcinogen for cancers of the skin, bladder, and lung. Emerging evidence also suggests that arsenic exposure may be a risk factor for breast and other cancers. Previous studies have been conducted in populations with endemically high levels of arsenic; however, the impact of chronic exposure to low levels is not known. Thus, the purpose of this study was to describe the arsenic exposure in Canadian adults and to evaluate the association between arsenic status and cancer risk.

Methods: The Canadian Health Measures Survey (CHMS) is an ongoing, nationally representative survey that collects self-reported and objective health measures, including biomonitoring data, from participants in biennial cycles. This analysis utilized total, inorganic and organic urinary arsenic biomarkers (µg/L) from Cycles 1-5 (2007-2017) of the CHMS. Incident cancers were ascertained through a linkage to the Canadian Cancer Registry. Arsenic exposure was described using weighted percentiles, and Cox proportional hazards regression models were used to estimate the association between arsenic exposure and cancer risk (overall and site-specific).

Results: This analysis included 10,000 individuals, aged 18-80. The median total urinary arsenic level was 7.5 ug/g of creatinine. Preliminary models showed that levels of inorganic arsenic exposure above the geometric mean (> 4.5 µg/L) were associated with a 3-fold increased risk of developing any cancer (HR = 3.58; 95%CI 1.63-7.85), compared to those with lower levels. Site-specific risk modelling is ongoing and preliminary results will be presented.

Significance: This study found inorganic arsenic exposure may be associated with an increased risk of developing cancer. To our knowledge, this study is the first to ascertain national estimates of arsenic exposure and provide insight on the relationship between arsenic exposure and cancer risk in Canadian adults. These findings are critical for developing population-level interventions to reduce the Canadian, and global, cancer burden.

Citation Format: Katherine Pullella, Vicky C. Chang, Shelley A. Harris, Anthony J. Hanley, John R. McLaughlin, Jan Lubinski, Steven A. Narod, Joanne Kotsopoulos. Examining the association between arsenic exposure and cancer risk among Canadian adults [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5895.
Abstract 5898: Bone morphogenic protein receptor 2 (BMPR2) as a potential germline driver in Juvenile Polyposis Syndrome (JPS)
Authors: MacFarland Suzanne P. , Greed Bridgid , Xie Michael H. , Brodeur Garrett M. , Cramer Zvi , Dent Maiah H. , Duvall Melani M. , Hamilton Kathryn E. , Howe James , Karakasheva Tatiana , Mamula Petar , Lengner Christopher
Published: June 2022

Summary: Juvenile Polyposis Syndrome is a pediatric cancer predisposition syndrome for which approximately half of patients do not have a known germline driver; we propose BMPR2 as a possible germline driver of disease in mutation-negative JPS.

Background: Juvenile Polyposis Syndrome (JPS) is a cancer predisposition syndrome characterized in some cases by inactivating germline mutations in BMPR1A or SMAD4. However, in 40-60% of patients the germline driver is unknown.

Methods: Through whole exome sequencing, an individual with high polyp burden and negative genetic testing for variants in SMAD4 and BMPR1A was found to have a potentially inactivating germline variant in BMPR2. Under an IRB-approved protocol, patient-derived, three-dimensional organoids were established from adjacent colon and polyp tissue of this individual, as well as an individual with a BMPR1A deletion and age- and sex-matched controls. Proliferation, metabolic activity, and protein expression of each line were measured by Western blot, Ki67 staining, EdU labelling, cell titer glo (CTG) assay. Results: Distinct phenotypic differences are evident between colonoids from an individual with a BMPR1A deletion and both the colon and polyp BMPR2 variant colonoids, such as increased crypt budding and Ki67 staining. CTG assay data indicate higher metabolic activity in BMPR2 variant colonoids in comparison to normal controls (p= 0.0104.) Through Western blot analysis, both BMPR2 colon and polyp colonoids showed decreased SMAD4 expression and phosphorylation.

Conclusions: These data demonstrate that clear phenotypic differences exist between colonoid lines established from normal controls, individuals with known JPS predisposition genes, and a novel candidate driver of disease, BMPR2. Proliferation and protein expression data suggest that both known and candidate genotypes are consistent with a hyperproliferative phenotype. In vitro data support BMPR2 as a candidate germline driver of the JPS phenotype, and additional study is ongoing regarding downstream pathway regulation in colonoids with a BMPR2 variant.

Citation Format: Suzanne P. MacFarland, Bridgid Greed, Michael H. Xie, Garrett M. Brodeur, Zvi Cramer, Maiah H. Dent, Melani M. Duvall, Kathryn E. Hamilton, James Howe, Tatiana Karakasheva, Petar Mamula, Christopher Lengner. Bone morphogenic protein receptor 2 (BMPR2) as a potential germline driver in Juvenile Polyposis Syndrome (JPS) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5898.
Abstract 5904: The clinical, behavioral, and demographic factors associated with the quality of life of cancer survivors
Authors: Chirinos Odette , Lee Eunkyung
Published: June 2022

The objective of this thesis is to assess the prevalence of low quality of life scores in adult cancer survivors. This thesis also intends to examine the predictors for low quality of life, separately for physical quality of life and mental quality of life. Possible predictors explored include diet quality, age at cancer diagnosis, race and ethnicity, body mass index, alcohol intake, physical activity, and more. The data source for this thesis is the National Health and Examination Survey 2005-2012. The sample population included 1244 cancer survivors 20 years or older that answered yes to the question “have you ever been told by a doctor or other health professional that had cancer or malignancy of any kind”? Both, mental and physical health, were determined based on the number of days physical and mental health was not good and dichotomized according to mean value. The four groups identified were low mental health and low physical health (n=148, 11.9%), low mental health and high physical health (n=239, 19.2%), high mental health and low physical health (n=143, 11.5%), and high mental health and high physical health (n=714, 57.4%). The diet quality was calculated using the Healthy Eating Index 2015 since it measures adherence to the Dietary Guidelines for Americans. The SAS Survey Analysis Procedures was used to account for the random multi-stage sampling of NHANES. On the other hand, ANOVA and chi-square were used to determine the possible predictors of low quality of life while multivariable logistic regression analysis was performed to examine the independent predictors of low physical and mental health with statistical significance was set at a two-sided p-value of <0.05. The results showed that age, economic status, and physical activity are predictors of low quality of life for adult cancer survivors. This indicates that future interventions for improving quality of life should be focused on increasing physical activity, especially in middle aged, obese, and economically disadvantaged cancer survivors.

Citation Format: Odette Chirinos, Eunkyung Lee. The clinical, behavioral, and demographic factors associated with the quality of life of cancer survivors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5904.
Abstract 5922: Clinical validation of a spectroscopic liquid biopsy for early detection of brain cancer
Authors: Cameron James M. , Brennan Paul M. , Antoniou Georgios , Butler Holly J. , Christie Loren , Conn Justin J.A. , Curran Tom , Gray Ewan , Hegarty Mark G. , Jenkinson Michael , Orringer Daniel , Palmer David S. , Sala Alexandra , Smith Benjamin R. , Baker Matthew J.
Published: June 2022

Diagnostic delays impact the quality of life and survival of patients with brain tumors. Earlier and expeditious diagnoses in these patients are crucial to reducing the morbidities and mortalities associated with brain tumors. A simple, rapid blood test that can be administered easily in a primary care setting to efficiently identify symptomatic patients who are most likely to have a brain tumor would enable quicker referral to brain imaging for those who need it most. Blood serum samples from 603 patients were prospectively collected and analyzed. Patients either had non-specific symptoms that could be indicative of a brain tumor on presentation to the Emergency Department, or a new brain tumor diagnosis and referral to the neurosurgical unit, NHS Lothian, Scotland. Patient blood serum samples were analyzed using the Dxcover® Brain Cancer liquid biopsy. This technology utilizes infrared spectroscopy combined with a diagnostic algorithm to predict the presence of intracranial disease. Our liquid biopsy approach reported an area under the receiver operating characteristic curve of 0.8. The sensitivity-tuned model achieves a 96% sensitivity with 45% specificity (NPV 99.3%) and identified 100% of glioblastoma multiforme patients. When tuned for a higher specificity, the model yields sensitivity of 47% with 90% specificity (PPV 28.4%). This simple, non-invasive blood test facilitates the triage and radiographic diagnosis of brain tumor patients, while providing reassurance to healthy patients. Minimizing time to diagnosis would facilitate identification of brain tumor patients at an earlier stage, enabling more effective, less morbid surgical and adjuvant care.

Citation Format: James M. Cameron, Paul M. Brennan, Georgios Antoniou, Holly J. Butler, Loren Christie, Justin J.A. Conn, Tom Curran, Ewan Gray, Mark G. Hegarty, Michael Jenkinson, Daniel Orringer, David S. Palmer, Alexandra Sala, Benjamin R. Smith, Matthew J. Baker. Clinical validation of a spectroscopic liquid biopsy for early detection of brain cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5922.
Abstract 5958: CD276/CD47-targeted antibody-drug conjugates to treat triple-negative breast cancers
Authors: Si Yingnan , Chen Kai , Kim Seulhee , Zhou Zhuoxin , Zhou Lufang , Liu X. Margaret
Published: June 2022

Triple-negative breast cancers (TNBCs) are highly aggressive, heterologous and frequently recurrent. The standard radiotherapy and chemotherapy have limited clinical benefits while the combined therapies or antibody-drug conjugates (ADCs) are promising to treat TNBC. The objective of this study was to develop a combined therapy, CD276/CD47-targeted ADCs, for TNBC treatment. Specifically, the CD276 and CD47 surface receptors were identified with immunohistochemistry staining of drug-sensitive and resistant patient tissues and confirmed in other assays. Two innovative monoclonal antibodies (mAbs), i.e. anti-CD276 mAb and anti-CD47 mAb that target the N-glycosylated extracellular domains (Leu29-Pro245 and Gln19-Glu141) of surface receptors, were developed using hybridoma technology. The top mAb clones with high surface binding, drug delivery and affinity were screened, produced in fed-batch culture, purified using liquid chromatography, and conjugated with payload mertansine (DM1). Flow cytometry, confocal microscopy, and in vivo imaging system (IVIS) showed that the CD276/CD47 ADCs effectively target human and mouse TNBC cells and xenograft models. The in vitro anti-cancer cytotoxicity was confirmed in multiple cell lines (MDA-MB-468, MDA-MB-231, 4T1), and in vivo evaluations using TNBC cell line-derived xenograft models and patient-derived xenograft (PDX) models demonstrated high anti-tumor efficacy and tumoral immunity of the dually targeted ADCs. In addition, whole blood analysis indicated that neither CD276 mAb nor CD47 mAb showed general immune toxicity in immunocompetent model. The maximal tolerated dose (MTD), pharmacokinetics (PK) and biodistribution will be further investigated. This study indicated that the ADCs dually targeting CD276 and CD47 have great potential to treat TNBCs.

Citation Format: Yingnan Si, Kai Chen, Seulhee Kim, Zhuoxin Zhou, Lufang Zhou, X. Margaret Liu. CD276/CD47-targeted antibody-drug conjugates to treat triple-negative breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5958.
Abstract 5977: Discovery and preclinical validation of [11C]AZ3391: A first in class blood-brain barrier permeable, subtype selective PARP-1 PET radioligand
Authors: Johnström Peter , Johannes Jeffrey , Pike Andy , Cselényi Zsolt , Schou Magnus
Published: June 2022

Positron emission tomography (PET) is a molecular imaging technique used to study the distribution of radiolabeled tracer molecules in vivo in a non-invasive fashion. We here report the discovery and preclinical validation of a novel PET tracer for PARP1. Its exquisite selectivity and high contrast in vivo opens up unique opportunities for exploring target engagement and relationships between drug exposure in plasma and PARP1 occupancy during clinical development of PARP1 inhibitors.

AZ3391 was identified as a PET radioligand candidate from our medicinal chemistry program investigating novel PARP1 selective inhibitors and radiolabeled with carbon-11 (t1/2=20.4 min) for a series of in vitro and in vivo imaging experiments.

Using in vitro autoradiography, dense binding of [11C]AZ3391 was observed in tissues known to be rich in PARP1 (eg tumour xenograft; NHP and human brain cerebellum). [11C]AZ3391 binding was completely abolished by the inclusion of a high concentration of Olaparib or AZD5305 in the incubate, thus demonstrating specific binding to PARP1 in vitro.

Following intravenous injection of [11C]AZ3391 in non-human primates, high binding was observed in organs known to express PARP1, eg brain, spleen and bone marrow. The radioactivity in these organs could be blocked in a dose-dependent manner after pre-treatment with increasing doses of AZD9574 (up to 0.05 mg/kg), thus demonstrating specific binding to PARP1 in vivo.

This set of preclinical data supports further development of [11C]AZ3391 as a tool to assess drug induced PARP1 engagement in patients.

Citation Format: Peter Johnström, Jeffrey Johannes, Andy Pike, Zsolt Cselényi, Magnus Schou. Discovery and preclinical validation of [11C]AZ3391: A first in class blood-brain barrier permeable, subtype selective PARP-1 PET radioligand [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5977.
Abstract 5992: Clonal evolutionary molecular mechanism of lymphatic metastasis in lung cancer
Authors: Guo Nanan , Zhao Hongmei
Published: June 2022

Background: Lung cancer approximately accounts for more than a third of cancers, approximately 787,000 in China annually, which the death rate is greatest and most lethal in all cancers. However, clonal evolutionary molecular mechanism of lymphatic metastasis in lung cancer remains largely unknown according to previously reported.

Methods: In this study, we systematically analyzed clonal evolution of 21 primary patients paired with lymphatic metastasis based on 500 gene-panel target sequencing and clinical characters. The two pipelines were performed to identify the subclones between primary and lymphatic metastasis on each patient, one is Pyclone, citup and Timespace, another is sciClone and clonevol. Furthermore, the genes associated with clonal evolutions were verified to be differentially mutated genes between primary and lymphatic metastasis in MSK-IMPACT 2017 cohort by fisher-test (P < 0.05).

Results: The clonal clusters were explored in 3 of 21 patients (P08, P52, P57) with lung cancer by first pipeline, and 4 (P02, P08, P14 and P84) by second pipeline. The 2 genes TET2 and BRCA2 were identified to be associated with lymphatic metastasis. The mutation frequency of TET2 and BRCA2 were 4.2% and 7.1% in patients with lymphatic metastasis in validation cohort. Conclusions: We described molecular features of lung cancer patients with lymphatic metastasis. TET2 and BRCA2 mutations promoted development and evolution of tumor cells, which were insightful for developing novel therapeutic strategies to inhibit the progress of tumors.

Keywords: Clonal Evolutionary, Molecular Mechanism, Lymphatic Metastasis, Lung cancer

Citation Format: Nanan Guo, Hongmei Zhao. Clonal evolutionary molecular mechanism of lymphatic metastasis in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5992.
Abstract 6010: Mouse hind limb tumor metastatic model to evaluate prophylaxis and treatment
Authors: Shuai Yao , Wang Chaomeng , Tong Ling , Liu Qiao , Xu Jie , Ji Qunsheng
Published: June 2022

We have established and optimized a robust tumor bone metastasis mouse model, which recapitulates the tumor metastasis progression from circulation to bone colonization, and provides windows for both prophylaxis and treatment. Specifically, we inoculated two breast cancer cell lines, JIMT-1 and MDA-MB-231, each with high metastatic potential, into mouse intra-caudal artery. By detection of steadily incremental bioluminescence signal, prophylaxis was observed to precede metastases, which were found exclusively on mouse hind limbs. These mouse models exhibited swollen hind limbs and even paralysis at late stage. Gross necropsy and pathology analysis confirmed the exclusive hind limb localization, the invasion and spreading of tumor cells to muscles and bone marrows, and the lesions in tibias and femurs. Our robust bone metastases models thus imitate the multiple stages of tumor metastasis, leading to exclusion bone metastasis and opening up windows for prophylaxis and treatment.

Citation Format: Yao Shuai, Chaomeng Wang, Ling Tong, Qiao Liu, Jie Xu, Qunsheng Ji. Mouse hind limb tumor metastatic model to evaluate prophylaxis and treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6010.
Abstract 6034: Characterization of a novel syngeneic cSCC mouse model to advance immunotherapy treatments
Authors: Rosario Alanis Enid Rodriguez , Rangel Roberto , Myers Jeffrey
Published: June 2022

Cutaneous Squamous Cell Carcinoma (cSCC) is the second most common malignancy diagnosed in Caucasians with an incidence of about one million cases per year. TP53 and p16 are the most common mutated genes in cSCC. Previous studies of cSCC have shown that patients with both p53 and p16 mutations have lower survival rates. There are currently no syngeneic mouse preclinical models to accelerate the efficacy of immunotherapy combinations to treat cSCC. Our laboratory has generated several genetically engineered mouse models (GEMMs) containing tissue-specific p53 and p16 gene mutations. Recently, we have immortalized three murine cSCC cell lines (cSCC-5690, cSCC-8006, and cSCC-8271) derived from spontaneous skin tumors from GEMMs. Therefore, we hypothesized that the double mutant cell line cSCC-5690 enhances cell survival, tumor aggressiveness, and has lower sensitivity to antibody immunotherapy. Protein and RNA expression of the cell lines were validated by western blotting and qRT-PCR assays. Sphere and soft agar assays were used to evaluate the stem cell population and tumorigenicity of the cell lines. Then, we examined the chemosensitivity with cisplatin through clonogenic assays. To further study the tumorigenicity of cells, we evaluated in vivo tumor growth on C57BL/6J immunocompetent mice by injecting subcutaneously the tumor cell lines. The three cell lines expressed different levels of keratin 14 (K14), but cSCC 5690 and 8006 expressed high levels of genes related to oncogenic pathways as well as mesenchymal markers. The cell line cSCC-5690 expressing mutant p53 R172H with p16 gene deletion, showed an enhanced sphere, and colony tumor formation and both tumorigenic cell lines (cSCC-5690, 8006) showed different sensitivities to cisplatin treatment. A higher tumor growth rate was observed with cSCC-5690 compared to cSCC-8006, while cSCC-8271 showed no tumor growth. Immunohistochemistry studies demonstrated that both cSCC-5690 and 8006 tumors expressed vimentin but only cSCC-5690 expressed K14. The most aggressive syngeneic tumor model (cSCC-5690) was used to evaluate the anti-tumor efficiency of anti-PD1, anti-CD47 antibodies, and STING-agonist. Preliminary results revealed a significant tumor response only to STING agonist (c-di-GMP) which activates the innate and adaptive immune response. Therefore, we were able to characterize all three cell lines and established a new mouse model of cSCC as a relevant platform for immunotherapy studies.

Citation Format: Alanis Enid Rodriguez Rosario, Roberto Rangel, Jeffrey Myers. Characterization of a novel syngeneic cSCC mouse model to advance immunotherapy treatments [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6034.
Abstract 6092: The dual roles of C-X-C MOTIF chemokine LIGAND 10 in pediatric osteosarcoma
Authors: Chen Xiang , Clement Margaret , Hicks John , Man Tsz Kwong
Published: June 2022

Osteosarcoma is the most common malignant bone tumor in children and young adults. Despite the use of multi-agents chemotherapy and aggressive surgery, the survival of patients with osteosarcoma remains at 60% over the past three decades. In addition, the survival of patients who develop metastasis is dismal and often less than 25%. We have previously reported that serum C-X-C motif chemokine ligand 10 (CXCL10) is associated with survival in a large cohort of osteosarcoma patients. CXCL10 is an interferon-induced chemokine that binds to its cognate receptor CXCR3 to elicit its cellular effects via both autocrine and paracrine signaling. CXCL10 can promote tumor cell migration and metastasis via autocrine signaling, while it recruits CXCR3+ immune cells to exert a tumoricidal effect via paracrine signaling. However, little is known about the effect of circulating CXCL10 in cancer. To understand the role of CXCL10 in the pediatric bone tumor, we showed that a number of osteosarcoma cell lines expressed the CXCR3 protein. The addition of exogenous CXCL10 promoted tumor cell migration of three osteosarcoma cell lines but did not affect cell proliferation. However, the migratory effect of osteosarcoma cells is not specific to CXCL10, as other CXCR3-associated chemokines (CXCL4, CXCL9, and CXCL10) exerted similar effects. Using an orthotopic xenograft mouse model, we demonstrated intravenous injection of CXCL10 increased the development of pulmonary metastases of osteosarcoma. In contrast, expression profiling of osteosarcoma primary tumors showed that elevated expression of CXCL10 and CXCR3 was associated with a better prognosis, suggesting that a paracrine effect of CXCL10 by recruiting CXCR3+ immune cells to the tumor site. This notion is supported by our immune cell population deconvolution results, indicating that the predominant infiltrating immune cell types are macrophages and CD8+ T cells. Taken together, our results suggest a model in which CXCL10 may play both pro-tumor and anti-tumor roles in osteosarcoma. While a high level of exogenous or circulating CXCL10 may promote tumor cell migration and metastasis, the chemokine can also recruit CXCR3+ immune cells to exert a favorable prognostic effect. Further characterization of the sources of CXCL10 in the tumor site and circulation may lead to the development of a therapeutic approach that promotes CXCL10’s anti-tumor effects and minimizes its pro-tumor effects to improve the survival of osteosarcoma patients.

Citation Format: Xiang Chen, Margaret Clement, John Hicks, Tsz Kwong Man. The dual roles of C-X-C MOTIF chemokine LIGAND 10 in pediatric osteosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6092.
Abstract 6098: Analysis of the impact of ZEB1 expression on pathway enrichment and immune cell abundance in prostate cancer
Authors: Chaves Luiz Paulo , Caliari André Luiz , Lautert-Dutra William , Silva Francisco Cesar Souza , Melo Camila Morais , Saggioro Fabiano Pinto , Borges dos Reis Rodolfo , Squire Jeremy Andrew
Published: June 2022

Prostate cancer (PCa) reactivates a latent embryonic program called epithelial-mesenchymal transition during disease progression. The mesenchymal phenotype contributes to invasion, metastasis and therapy response variation. In this study, we characterize the downstream effects of ZEB1 expression based on cohort dichotomization of quartiles of expression comparing the 25% of tumors with high ZEB1 expression to the 75% of tumors with lower ZEB1 expression. Differential gene expression (DGE), gene set enrichment analysis (GSEA) and immune cell abundance prediction were accessed using PCa expression data from TCGA (n=488). ZEB1 high status based on gene expression was stratified based on ZEB1 expression values higher than the 3rd expression quartile value (n=122). Contingency analysis showed that PTEN loss is more frequent in the ZEB1 high group (Chi-square, p=0.0005) which supports previously presented data from our group that showed an inverse correlation between ZEB1 and PTEN protein expression but showed no difference between the groups regarding presence of the TMPRSS2-ERG fusion. DGE showed that more than 13,000 genes are differentially expressed in the ZEB1 high group while GSEA showed that signaling pathways such as HALLMARK TGF BETA, HALLMARK IL2 STAT5, HALLMARK INFLAMMATORY RESPONSE and HALLMARK NOTCH were upregulated in the ZEB1 high group compared to the low group. Yet inflammatory response related pathways were observed enriched in the ZEB1 high group, CIBERSORT analysis showed that effector immune cells such as CD8+ T cells and T helper cells (Holm-Sidak, p=000003 and p=0.000613, respectively) were enriched in the ZEB1 low group in comparison to the ZEB1 high group, which supports previous studies that suggest that ZEB1 cooperates with PTEN loss and TMPRSS2-ERG fusion to promote an immune evasive phenotype in PCa. Collectively, these findings demonstrate that gene expression changes associated with increased ZEB1 expression may involve clinically relevant pathways such as modulation of the tumor immune microenvironment and NOTCH-dependent tumor progression.

Citation Format: Luiz Paulo Chaves, André Luiz Caliari, William Lautert-Dutra, Francisco Cesar Souza Silva, Camila Morais Melo, Fabiano Pinto Saggioro, Rodolfo Borges dos Reis, Jeremy Andrew Squire. Analysis of the impact of ZEB1 expression on pathway enrichment and immune cell abundance in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6098.
Abstract 6101: K6-K14 keratin fusion variant (K6-K14/V7) in OSCC cells promotes tumor microenvironment remodeling through activating inflammatory CAFs
Authors: Kuo Brian Yu-Ting , Lin Yen-Ting , Ravichandran Senthilkumar , Sheu Jim Jinn-Chyuan
Published: June 2022

Keratin intermediate filament is one major cytoskeleton architecture in epithelial cells, providing necessary mechanical support to maintain cell shape and tissue integrity. In a previous study, we discovered the existence of a novel K6-K14 keratin fusion variant (K6-K14/V7) in oral squamous cell carcinoma (OSCC) cells which can promote cancer stemness and aggressiveness. However, the underlying mechanism remains to be elucidated. Here, we report that overexpression of K6-K14/V7 in OSCC cells can influence the mechanical properties of the cells, leading to cell protrusion and enhanced extracellular vesicle (EV) secretion. Consequently, the expression levels of inflammatory cytokines (e.g. IL-6, TGF-[[Unsupported Character - Symbol Font β]]) in the culture medium can be also increased. Normal oral fibroblast cells co-cultured with K6-K14/V7-expressing OSCC cells can be converted into a pro-inflammatory type of cancer-associated fibroblasts (iCAFs). Those findings suggest the ability of K6-K14/V7-expressing OSCC cells in remodeling tumor microenvironment. In this study, we will discuss how K6-K14/V7-expressing OSCC cells educate iCAFs to promote cancer cell migration and invasion via intercellular communication by EVs. The key signaling cascades and the associated regulatory networks will be also discussed. Our study, therefore, uncovers the pro-tumorigenic functionality of K6-K14/V7 by creating an inflammatory tumor microenvironment.

Citation Format: Brian Yu-Ting Kuo, Yen-Ting Lin, Senthilkumar Ravichandran, Jim Jinn-Chyuan Sheu. K6-K14 keratin fusion variant (K6-K14/V7) in OSCC cells promotes tumor microenvironment remodeling through activating inflammatory CAFs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6101.
Abstract 6116: Tumor immune environment in liver metastasis of colorectal cancer and its correlation with recurrence free survival
Authors: Zhou Jiamin , Xu Weiqi , Wang Miao , Zhang Siqi , Zhang Ding
Published: June 2022

Introduction: The liver is a common site of metastases of colorectal cancer. Tumor infiltrating lymphocytes (TILs) play an important role in the process of colorectal cancer liver metastasis (CRCLM). A comprehensive investigation into TILs will provide more prognostic information for patients with colorectal liver metastases after liver resection.

Methods: 15 CRC patients with liver metastases from our hospital were enrolled in the study. The density and percentage of the TILs [CD8+, CD3+, CD4+, CD20+, tumor associated macrophages (TAMs), dim CD56+NK, bright CD56+NK, PD-1+etc.] in the liver metastasis samples from operation were analyzed by multiplex immunofluorescence technique (3DMedcines Inc.). The level of the density and percentage of immune cells were determined by dichotomy method. The correlation between recurrence free survival (RFS) and immune cells were analyzed using the Kaplan-Meier curve and Log-rank test.

Results: The existence of dim CD56+ NK cells in the stroma was associated with significant improvement in RFS (p=0.0461). The median RFS was 26.1 months (95% CI, 14.9 to 33.5) with the existence of dim CD56+ NK cells in the stroma versus 1.7 months (95% CI, -18.0 to 32.4) without dim CD56+ NK cells in the stroma. Nevertheless, dim CD56+ NK cells in the tumor-enriched segments was not significantly associated with improved outcome. The existence of bright CD56+ NK cells in the stroma was associated with significant improvement in RFS (p=0.0307) but not in the tumor-enriched segments. The median RFS was 34.6 months (95% CI, 15.0 to 35.2) with the existence of bright CD56+ NK cells in the stroma versus 7.9 months (95% CI, -5.3 to 23.1) without bright CD56+ NK cells in the stroma. However, low FoxP3+ cells in the tumor-enriched segments (p=0.0037) was significantly associated with improved outcome but not in the stroma. The median RFS was 36.5 months (95% CI, 23.1 to 40.1) with low FoxP3+ cells versus 8.2 months (95% CI, 0.5 to 21.8) with high FoxP3+ cells in the tumor-enriched segments. Low CD3+CD4+FoxP3+ cells (p=0.074) and the existence of PD-L1+CD68+ cells (p=0.0524) in the tumor-enriched segments showed a trend towards better RFS. No significant correlation was achieved for CD8+, CD3+, CD3+CD4+, CD20+, CD68+CD163+, CD68+CD163-, PD-1, PD-1+CD8+ cells either in the tumor-enriched segments nor in the stroma.

Conclusion: We identified that CD56+ NK cells in stroma are associated with improved outcome in CRCLM. Nevertheless, FoxP3+ cells are associated with poor prognosis in CRCLM.

Citation Format: Jiamin Zhou, Weiqi Xu, Miao Wang, Siqi Zhang, Ding Zhang. Tumor immune environment in liver metastasis of colorectal cancer and its correlation with recurrence free survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6116.
Abstract 6135: Microbiome profiling through the various gastrointestinal environment of gastric cancer
Authors: Lee Jieun , Shin Sunguk , Lee Seung-been , Yoo Yieri , Lee Sangjun , Kang So Hyun , Lee Eunju , Park Young Suk , Ahn Sang-Hoon , Park Kyoung Un , Kim Hyung-Ho , Kwon Nak Jung , Suh Yun-Suhk
Published: June 2022

The association between cancer and microbiome dysbiosis across anatomically related multiple body sites has not been comprehensively investigated. The purpose of our study is to profile microbial diversity and composition through the various gastrointestinal environment of gastric cancer (GC). We performed V3-V4 16S rRNA gene sequencing analysis for matched samples of gastric tumor, normal gastric mucosa, gastric juice and stool from 30 GC patients. Amplicon sequence variant (ASV) profile was compared among the four body sites at genus level. In this study, we found that mean alpha diversity was lowest in normal gastric mucosa and stool exhibited the largest amount of alpha diversity compared with others. Beta-diversity analysis showed significant differences in microbiota composition for each sample. The microbiome dysbiosis was significantly independent in gastrointestinal environment of gastric cancer. Helicobacter abundance in tumor tissue was significantly lower than in matched normal tissue and gastric juice while the trend was opposite for Lactobacillus. Additionally, the level of Helicobacter was considerably lower in patients with lymphatic invasion. The bacterial community that significantly correlated with tumor samples compared to normal mucosa, gastric juice, and stool were 49, 27, and 11 genus, respectively. Lactobacillus and Delftia had higher abundance and Rothia and Collnsella had lower abundance in tumor tissue compare with normal mucosa. Especially, Delftia was seen only in the tumor tissue not normal gastric mucosa, gastric juice and stool. Pentose phosphate pathway was significantly enriched in tumor tissue and normal mucosa. There is a unique microbiome pattern through the various gastrointestinal environment of gastric cancer. Our analysis shows enriched Delftia abundance only in the tumor tissue except other sample type.

Citation Format: Jieun Lee, Sunguk Shin, Seung-been Lee, Yieri Yoo, Sangjun Lee, So Hyun Kang, Eunju Lee, Young Suk Park, Sang-Hoon Ahn, Kyoung Un Park, Hyung-Ho Kim, Nak Jung Kwon, Yun-Suhk Suh. Microbiome profiling through the various gastrointestinal environment of gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6135.
Abstract 615A: HuGEMM࣪ modelling toxicology of species-specific immune checkpoint inhibitors
Authors: He Daniel XF
Published: June 2022

Background: Immune checkpoint inhibitors, including anti-PD-1 and anti-CTLA-4 monoclonal antibodies, have led to impressive clinical outcomes in treating certain cancers. However, the overall success rate of immunotherapeutic drugs remains low, due to poor efficacy (cancer immunotherapeutic effect [CITE]) or unexpected safety risks such as immune-related adverse events (irAEs), underlining the importance of predictive preclinical animal models. Notably, most immunotherapies cannot be modeled in xenograft tumor models, due to their immunodeficient phenotype, or syngeneic models, as most biologics are usually species-specific. Given these limitations, genetically engineered humanized mouse models (HuGEMM࣪) and syngeneic cell lines (HuCELL࣪), where human drug target gene(s) replace the original mouse ortholog(s), have been broadly used to assess species-restricted biologics for CITE. Since most irAEs in biologics are on-target toxicities, it is potentially possible to model the irAEs of biologics using non-tumor bearing HuGEMMs. In this study, the PD-1/CTLA-4 double knock-in (dKI) HuGEMM is used as an example to provide more insights into irAE evaluation in preclinical models.

Methods: The PD-1 and CTLA-4 HuGEMM in BALB/c backgrounds were developed by substituting the mouse extracellular domains with their human counterparts, followed by crossbreeding to create dKI mice for both targets. The juvenile dKI mice (age 4-5 weeks) were treated with the vehicle control, anti-human PD-1, and a combination therapy of anti-human PD-1/anti-human CTLA-4. The models were monitored for toxicity parameters, including general welfare, body weight, mortality, gross pathology (arthritis, skin inflammation, and colitis scoring), and histopathology of different organs at study termination.

Results: No body weight loss was observed throughout the study in all treatment groups. No mortality was observed in models treated with a monotherapy, while in the combination group (anti-human PD-1/anti-human CTLA-4), five mice displayed lethargy and tachypnea symptoms, and four mice experienced severe adverse effects. Per gross pathology analysis, combination treatment caused significant arthritis in the feet of the mice at study end. Histopathology analysis of the combination treatment group samples demonstrated colon epithelia damage and heart lymphocyte infiltration at termination. These observations for toxicity or irAE profiles are in line with those observed in patients undergoing the same treatment regimen.

Conclusions: HuGEMM engineered to express human targets could be used in preclinical studies to test for assessing irAE of biologics.

Citation Format: Daniel XF He. HuGEMM࣪ modelling toxicology of species-specific immune checkpoint inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 615A.
Abstract 6160: Tie2-mediated AMPK activation by ferritin-based protein C nanoparticles inhibits advanced prostate cancer development through induction of vasodilation
Authors: Lee You Mie , Jeong Ji-Hak , Jang Hyunha , You Jihye
Published: June 2022

Prostate cancer (PCa) is one of the most common malignancies and the second-leading cause of cancer-related mortality in men in Western countries. Androgen deprivation therapy (ADT) is an initial systemic therapy for advanced PCa, but almost all cancer eventually becomes castration resistant. Recently, we introduced ferritin-based protein C nanoparticles (PCNs), known as TFG and TFMG, which enhance normalization of the tumor vasculature. However, the exact underlying mechanism how PCNs induces AMP-activated protein kinase (AMPK) activation and anti-tumor activity in castration-resistant prostate cancer (CRPC) remains unknown. Here, we investigated the role of PCNs for CRPC development in endothelial cells. We found that TFMG inhibits prostate cancer castration resistance by activating AMPK through the Tie2-mediated signaling pathway. TFMG treatment increased AMPK phosphorylation at Thr172 and acetyl-CoA carboxylase (ACC) at Ser79 in EA.hy926 cells. TFMG treatment also significantly increased the phosphorylation of endothelial nitric oxide synthase (eNOS) with inducing nitric oxide (NO) production in EA.hy926 cells. In addition, TFMG treatment induced the vasodilatory effect in isolated rat mesenteric resistance arteries (MRAs), which was blocked by NO synthase inhibitor L-NAME. Interestingly, these effects were all abolished by pretreatment with Compound C (an AMPK inhibitor), rebastinib (selective Tie2 Inhibitor), AMPK knockdown using siRNA, or dominant negative AMPKα recombinant adenovirus. Finally, we found that TFMG functionally abrogates CRPC development in mouse model. In summary, TFMG exerted vasodilatation through activating Tie2/AMPK/eNOS signaling in endothelial cells, which leads to the overcoming vasoconstriction induced ADT. This study provides the potential value of TFMG in vasodilation of blood vessels leading suppression of CRPC development.

Citation Format: You Mie Lee, Ji-Hak Jeong, Hyunha Jang, Jihye You. Tie2-mediated AMPK activation by ferritin-based protein C nanoparticles inhibits advanced prostate cancer development through induction of vasodilation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6160.
Abstract 6192: ERK/AKT pathways may be the wrong effectors for KRAS signaling inhibition in colorectal cancers
Authors: Cheah Peh Yean , Lam Kuen Kuen , Tang Choong Leong , Tan Emile , Wong Siew Heng
Published: June 2022

Colorectal cancer (CRC) is the third highest incident cancer worldwide and a major cause of cancer death. KRAS protein gain-of-function point mutations play major roles in colorectal tumorigenesis. However, direct inhibition of KRAS is challenging as its protein structure is inadmissible to inhibitor docking. Efforts have been diverted to inhibit downstream RAF/MEK/ERK and PI3K/Akt signaling. Nevertheless, none of these inhibitors has progressed to clinical use in CRC despite extensive trials over the past few decades. We examined levels of phospho-ERK1/2(T202/Y204) and phospho-Akt1/2/3(S473) in human colorectal tumor and matched mucosa at least 10 cm away with quantitative near infrared western blot and immunofluorescence imaging. Surprisingly, 75.5% (25/33) of tumors has lower or equivalent phospho-ERK1/2 and 96.9% (31/32) of tumors has lower phospho-AKT1/2/3 compared to matched mucosa, irrespective of KRAS mutation status and tumor stage. In contrast, we discovered KRAS-dependent SOX9 upregulation in 28 of the 31 (90.3%) tumors. CPTAC phosphoproteomics data similarly showed significantly lower phospho-ERK1/2 levels in tumor compared to matched mucosa. Analysis of transcriptome data from TCGA and GEO series GSE95132 also showed no activation of RAF/MEK/ERK signaling in CRC tumors. These data indicate that the pursue of targeted therapy inhibiting RAF/MEK/ERK and PI3K/Akt signaling is unlikely to be effective in sporadic CRC and possibly harmful as phospho-ERK1/2 are highly expressed in the matched mucosa suggesting a role in normal colonic homeostasis. The widespread use of RAF/MEK/ERK or PI3K/Akt signaling downregulation as a biomarker in drug testing on CRC cell lines needs to be re-evaluated. The data suggest further that SOX9 could be an alternative effector and biomarker of KRAS signaling.

Citation Format: Peh Yean Cheah, Kuen Kuen Lam, Choong Leong Tang, Emile Tan, Siew Heng Wong. ERK/AKT pathways may be the wrong effectors for KRAS signaling inhibition in colorectal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6192.
Abstract 6214: Detecting and characterizing extrachromosomal DNA in medulloblastoma at single-cell resolution
Authors: Chapman Owen , Sridhar Sunita , Larson Jon D. , Wechsler-Reya Robert J. , Mesirov Jill , Chavez Lukas
Published: June 2022

Extrachromosomal circular DNA (ecDNA) is an important driver of aggressive cancers, including medulloblastoma (MB), the most common malignant pediatric brain tumor. To assess the clinical import of ecDNA in MB, we applied computational methods to detect ecDNA to the genomes of a cohort of 468 MB patients and 31 MB model systems. Among patients, ecDNA was detected in 18% of tumors and carried threefold greater risk of mortality. Between sequential patient biopsies at initial diagnosis and subsequent relapse, we observed frequent structural variation at ecDNA loci and generation of new ecDNA sequences. Among model systems, ecDNA was found in 19 of 31 genomes (61%). Although ecDNA was far more prevalent among MB models than patients, the ecDNA genomic sequence was conserved between most patient-derived xenograft (PDX) models and their origin human tumors. EcDNA promotes high oncogene copy number, accelerated evolution of drug resistance and poor prognosis. Mechanistically, these behaviors are hypothesized to be consequences of intratumoral heterogeneity of ecDNA copy number; however, there are currently no methods to quantify the abundance and distribution of ecDNA across single cells in a tumor. We have previously identified a primary TP53-mutant SHH medulloblastoma tumor, which harbors multiple distinct lineages of ecDNA originating from different chromosomes. We have now simultaneously analyzed the accessible chromatin and gene expression in single cells of the human tumor and of a orthotopic mouse xenograft (PDX) derived for this tumor, using multiome single-cell ATAC-seq and gene expression (10X Genomics). By measuring scATAC-seq read depth at the ecDNA loci relative to the rest of the genome, we estimate ecDNA copy number in single cells. We observe substantial variation in the ecDNA copy number of cells in both the human tumor and the PDX, and strong association with transcription of ecDNA-amplified genes. We anticipate that this approach will illuminate mechanisms of ecDNA-mediated drug resistance, by revealing the sensitivity and population dynamics of ecDNA-containing cells under treatment pressure.

Citation Format: Owen Chapman, Sunita Sridhar, Jon D. Larson, Robert J. Wechsler-Reya, Jill Mesirov, Lukas Chavez. Detecting and characterizing extrachromosomal DNA in medulloblastoma at single-cell resolution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6214.