bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5935: Malignant risk assessment of clinical and histological subtypes of lichenoid mucositis with dysplasia
Authors: Lin Iris , Kim Oh Run , Rock Leigha , Zhang Lewei , Rosin Miriam , Guillaud Martial , Laronde Denise
Published: June 2022

Introduction: Oral lichen planus (OLP), a common, chronic inflammatory condition, is categorized as potentially malignant by the World Health Organization. However, some hypothesize that only lichenoid lesions with dysplasia (LLD) are at risk of progression to cancer. Others have proposed subtypes of LLD, with differing malignant risks. Objectives: To determine if subtypes of LLD exist, based on clinical and histological features, and to determine the proportion of malignant progression in these subtypes. Methods: A nested case control study was conducted using tissue samples and longitudinal data of patients enrolled in the Oral Cancer Prediction Longitudinal study. Inclusion criteria were a biopsy confirmed diagnosis of lichenoid mucositis with dysplasia, no history of head and neck cancer, at least 5 years of follow-up, and accessible clinical photographs and bio-banked tissue. A chart review was conducted, and photographs reviewed to assess for lesion features characteristic of OLP. Histological sections were examined for lichenoid inflammation, and immunohistochemical staining for collagen IV was used to evaluate the integrity of the basement membrane (BM). Two subtypes of LLD were proposed: 1) a primary lichenoid and secondary dysplastic (L1D2) type defined as a lesion with a reticular and bilateral clinical presentation, subepithelial inflammation, and evidence of BM degeneration, and 2) a primary dysplastic and secondary lichenoid type (D1L2), which may present unilaterally or bilaterally, clinically resemble OLP or leukoplakia, and exhibit subepithelial inflammation with or without evidence of BM degeneration. Association with outcome, which was defined as progression to severe dysplasia, carcinoma in situ, or squamous cell carcinoma, was tested using Fisher’s Exact test. Results: All progressing cases had a non-homogenous clinical presentation (P=0.005) and were more apt to change (in size, texture, and/or appearance) over time (P=0.002). A reticular, smooth appearance was more common in non-progressing cases (P=0.007, P=0.016, respectively). 30% of cases in the D1L2 subtype progressed to malignancy, compared to none in the L1D2 group, though this finding did not reach statistical significance (P=0.081). Conclusion: LLD with a non-homogenous, non-reticular clinical presentation are at greater risk of progression, and a high proportion of cases exhibiting primary dysplastic features progressed to malignancy, indicating that LLD should be under close clinical follow-up.

Citation Format: Iris Lin, Oh Run Kim, Leigha Rock, Lewei Zhang, Miriam Rosin, Martial Guillaud, Denise Laronde. Malignant risk assessment of clinical and histological subtypes of lichenoid mucositis with dysplasia [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 5935.
Abstract 5938: Administering colorectal cancer education via an inflatable colon with the underrepresented and underserved population of Arizona
Authors: Jones Desiree , Ruiz Gabriela , Contreras Juan , Herrera Rene D. , Lee Amanda A. , Yellowhair Monica , Hatcher Jennifer
Published: June 2022

Background: The state of Arizona has a diverse population, with the Latino (31.7%) and American Indian/Alaskan Natives (AI/AN) (5.3%) populations higher than the national average of 18.5% and 1.3%, respectively. The colorectal cancer (CRC) screening rates among Latinos is 54.8% and AI/AN is 35%, compared to the overall Arizona screening rate of 62.6%, potentially contributing to higher CRC mortality rates in these groups.

Objective: Utilizing an inflatable colon to measuring CRC knowledge and intent to screen among Arizona’s diverse populations by administering a pre/post test.

Methods: An inflatable colon with signs in both English and Spanish describes the progression of CRC. Spanish-speaking health educators and culturally tailored flip charts geared toward the AI/AN population are utilized at community events throughout Arizona. Survey distribution measured pre- and post-CRC knowledge and intent to screen following a walking tour of the inflatable colon. Additional efforts include the production of a virtual tour of the giant colon in English and Spanish as an educational tool easily accessible on the UArizona Cancer Center YouTube page.

Results: Convenience sampling is utilized, and participants are recruited at outreach events where the inflatable colon is set up (n=15). Preliminary findings show that individuals increased their knowledge about CRC (76.9% vs 100%), understanding the pathology of CRC (i.e., colon polyp, normal colon tissue vs. cancerous tissue, etc.) (53.8% vs 100%), and CRC screening recommendations (30.8% vs 91.7%).

Conclusions: Colorectal cancer education is reaching diverse populations in Arizona. CRC education is delivered in both English and Spanish, and additional CRC educational tools such as the culturally tailored colon health flipbook for AI/AN population are utilize. The use of the inflatable colon has increased CRC knowledge, CRC pathology, and CRC screening options.

Citation Format: Desiree Jones, Gabriela Ruiz, Juan Contreras, Rene D. Herrera, Amanda A. Lee, Monica Yellowhair, Jennifer Hatcher. Administering colorectal cancer education via an inflatable colon with the underrepresented and underserved population of Arizona [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 5938.
Abstract 5942: Vitamin D, calcium supplement use and the risk of breast cancer in BRCA1 and BRCA2 mutation carriers: A case-control study
Authors: Guyonnet Emma , Kim Shana J. , Zhang Cindy X. , McCuaig Jeanna , Armel Susan , Narod Steven A. , Kotsopoulos Joanne
Published: June 2022

Background: Use of vitamin D and calcium-containing supplements are associated with a decreased risk of breast cancer among women in the general population. Whether this association exists among women at a high-risk due to a BRCA1 or BRCA2 mutation is not known. Thus, we conducted a case-control study to evaluate the association between vitamin D and/or calcium supplement use and the risk of breast cancer among BRCA mutation carriers.

Methods: BRCA mutation carriers enrolled in a longitudinal study, that collected information on cancer history and lifestyle factors, were invited to complete a supplemental questionnaire on past supplement use. Vitamin D and calcium supplement use were categorized as never or ever use. Total average daily vitamin D and calcium use was stratified as never, moderate, or high intake based on tertiles. Unconditional logistic regression was used to estimate the odds ratio (OR) and 95% confidence intervals (CI) of invasive breast cancer associated with supplement use. Analyses were stratified by BRCA mutation type and menopausal status at breast cancer diagnosis.

Results: This study included 134 cases and 276 controls. Compared to never users, ever vitamin D or calcium-containing supplement use was associated with decreased odds of breast cancer (OR=0.54; 95%CI 0.31, 0.91; P=0.02 for vitamin D; OR=0.61; 95%CI 0.36, 1.05; P=0.07 for calcium). The protective effect of vitamin D and calcium was stronger for those with the highest intake of each supplement (OR=0.44; 95% CI 0.22, 0.89; P=0.02 for vitamin D; OR=0.55; 95% CI 0.28, 1.08; P=0.08 for calcium), compared to never use. For vitamin D use, findings were significant among premenopausal women while for calcium use, the inverse associations were stronger among postmenopausal women. Findings were also significant for BRCA1 mutation carriers only, however, stratified analyses were limited by small strata.

Conclusions: Although based on a relatively small sample, our findings suggest a potential protective effect of vitamin D and calcium-containing supplement use for breast cancer among high-risk women. Future studies with larger samples and prospective study design are warranted.

Citation Format: Emma Guyonnet, Shana J. Kim, Cindy X. Zhang, Jeanna McCuaig, Susan Armel, Steven A. Narod, Joanne Kotsopoulos. Vitamin D, calcium supplement use and the risk of breast cancer in BRCA1 and BRCA2 mutation carriers: A case-control study [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 5942.
Abstract 596: IOH-001, a novel CD47/PD-L1 bispecific antibody, enhances anti-tumor activity in solid tumors
Authors: Kim Jeong-kook , Jeon A-Ra , Park Jihyun , Choi Ji Yea , Park Ji Eun , Song Sun Kwang , Choi Ji Hye , Shin Heewook , Lee Ji Hye , Yun Ji Hye , Ahn Yoen Hee , Kim Heung Tae
Published: June 2022

Background: The survived cancer cells after cytotoxic chemotherapy are known to promote immune evasion phenotype by increasing the expression of PD-L1 and CD47. Targeting of CD47 and PD-L1 is predicted to overcome immune evasion by coordinately restoring each innate and adaptive immunity. IOH-001, a CD47/PD-L1 dual-targeting bispecific antibody, has been shown to activate immune cells in tumor microenvironment by blocking PD-1/PD-L1 signals that inhibit cytotoxic T-cells, while also blocking interaction with CD47/SIRPa between cancer cells and macrophages. We have investigated whether IOH-001 inhibits tumor growth via co-targeting tumor cells or not.

Methods: To assess the function of IOH-001, a series of in vitro functional assays including cell surface binding, antibody-dependent cellular cytotoxicity (ADCC), phagocytosis assays and mixed lymphocyte reaction (MLR) assay were performed compared with the parent antibody. In vivo efficacy of IOH-001 was tested in colon cancer mouse models with human genes KI.

Results: IOH-001 has been shown to be bound strongly to various types of PD-L1/CD47-expressing cancer cells including the ones in solid and hematological cancers. In most cancer cells, IOH-001 was demonstrated to have a lower EC50 than the parental antibodies and to be bound in a dose-dependent manner as well. Interestingly, IOH-001 also has shown the selectivity to bind only to cancer cells even under the conditions of co-culturing RBC and cancer cells. IOH-001 has induced phagocytosis of cancer cells by human blood CD14+ monocyte-derived macrophages. Since IOH-001 is an IgG1 type antibody, ADCC and IFN-r expression have been increased compared with the parental antibodies. Consistent with the in vitro data, IOH-001 has more strongly suppressed tumor growth than the combination treatment of the parental antibodies in a dose-dependent manner in syngeneic animal models. Moreover, the tumor of the CR mouse has been identified not to be re-generated in the re-challenge model.

Conclusion: IOH-001, dual-blockage of anti-CD47 and anti-PD-L1, has shown the benefits in treating some solid tumors. Bispecific antibody IOH-001 is more likely to work better in targeting tumor cells than the combination of anti-CD47 and anti-PD-L1. Preclinical efficacy results of IOH-001 provide a strong rationale for assessing therapeutic potential in clinical studies.

Citation Format: Jeong-kook Kim, A-Ra Jeon, Jihyun Park, Ji Yea Choi, Ji Eun Park, Sun Kwang Song, Ji Hye Choi, Heewook Shin, Ji Hye Lee, Ji Hye Yun, Yoen Hee Ahn, Heung Tae Kim. IOH-001, a novel CD47/PD-L1 bispecific antibody, enhances anti-tumor activity in solid tumors [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 596.
Abstract 5969: Intra-tumoral coagulation is significantly associated with EMT, angiogenesis, and worse patient survival in gastric cancer
Authors: Oshi Masanori , Sarkar Joy , Wu Rongrong , Yan Li , Matsuyama Ryusei , Endo Itaru , Takabe Kazuaki
Published: June 2022

Background: Bleeding is one of the most commonly observed clinical signs of gastrointestinal cancer. Coagulation regulates inflammation and angiogenesis in cancer, and is associated with tumor development and metastasis. To date, there has been no measures to quantify the state of intra-tumoral coagulation. We hypothesized that intra-tumoral coagulation as quantified using coagulation score is associated with angiogenesis and worse prognosis in gastric cancer.

Methods: The coagulation score was determined by the Gene set variation analysis (GSVA) algorithm as GSVA score of the “HALLMARK_COAGULATION” in the MSigDB hallmark gene sets collection. Transcriptome data of 807 gastric cancer patients of TCGA_STAD and GSE84437 cohorts were used for the analysis.

Results: We found that high coagulation score was significantly associated with high AJCC T-category (p = 0.018), lymph node metastasis status (p = 0.036), AJCC stage (p = 0.006) in the TCGA cohort. The association of the score with staging were validated by GSE84437 cohort. Further, patients with high score were associated with worse disease-specific survival and overall survival (OS) (p = 0.019 and 0.011, respectively) in the TCGA cohort, and OS in GSE84437 cohort (p = 0.012). Although the coagulation score was not associated with cell proliferation-related nor immune-related signaling, gastric cancer with high coagulation score highly enriched epithelial mesenchymal transition (EMT), myogenesis, apical junction, transforming growth factor (TGF)-β signaling, and angiogenesis consistently in both cohorts (all false discovery rate<0.25). The coagulation score strongly correlated with angiogenesis score, and high coagulation was significantly associated with high infiltration of angiogenesis-related vascular cells, including endothelial cells, microvascular endothelial cells, lymphatic endothelial cells, and pericytes, calculated by xCell algorithm, as well as high expression of angiogenesis-related genes, including Endothelial cell marker, and vascular stability related genes.

Conclusion: Intra-tumoral coagulation assessed by coagulation score is associated with clinical outcomes of gastric cancer patients.

Citation Format: Masanori Oshi, Joy Sarkar, Rongrong Wu, Li Yan, Ryusei Matsuyama, Itaru Endo, Kazuaki Takabe. Intra-tumoral coagulation is significantly associated with EMT, angiogenesis, and worse patient survival in 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 5969.
Abstract 5970: Tumor vessel normalization by a novel anti-TIE2 antibody PMC-403 enhances the effectiveness of radiation therapy on cancer therapy
Authors: Lee Eun-Ah , Park Beom Yong , Kang Nuri , Park Cheon Ho , Lee Weon Sup
Published: June 2022

Purpose: The delivery of drugs and immune cells into the tumor microenvironment is limited due to abnormal tumor vessel which is characterized as low oxygen concentration, low perfusion rate and leakage. It leads to diminish the efficacy radiation therapy. Traditional anti-angiogenesis strategies attempt to reduce the tumor vascular supply, but their success is restricted by insufficient efficacy or development of resistance. To solve the problem, we try to using PMC-403, which can normalize the abnormal condition of tumor vessel.

Experimental procedures: Target selectivity: It was measured by Surface Plasmon Resonance (SPR) assay. Human TIE2 Cho-K1 cells or mouse TIE2 Cho-K1 cells were tested for species cross-reactive assay based on flow cytometry (FACS).In vitro studies: Human umbilical vein endothelial cells (HUVEC) were used for TIE2 signal pathway analysis with western blot assay and efficacy analysis with VEGF-induced vessel permeability assay.In vivo study: Glioblastoma model mice were employed for the assessment of tumor vessel normalizing activity of 1 mg/mouse PMC-403. CT26 colon cancer model mice were used to evaluate the anti-tumor efficacy of PMC403 (10 mg/kg) in combination with radiation therapy. Hypoxyprobe was used to measure the improvement of hypoxic conditions in tumor tissue.

Summary of data: PMC-403 has a nanomolar range of affinity for both human or mouse TIE2 expressing cells. This contributed to dose-dependent ANG1-like TIE2 and FOXO1 phosphorylation and inhibited VEGF-induced vascular leakage, thereby contributing to ANG1-like vascular normalization. Moreover, similar to ANG-1, p-VEGFR2 and p-VE-Cadherin levels were significantly reduced by PMC-403, suggesting that PMC-403 can normalize blood vessels. In addition, PMC-403 stabilized abnormal tumor blood vessels in the glioblastoma mouse model and significantly reduced tumor growth and tumor hypoxia in the combination treatment (PMC-403 + radiation therapy) compared to radiation therapy alone, although the antitumor effect of PMC-403 alone was not effective in the same condition model.

Conclusions: PMC-403, an anti-TIE2 monoclonal antibody, is a novel tumor vessel stabilizer. PMC-403 improved tumor microenvironment hypoxic condition that synergized with radiotherapy when it combined together for the anticancer therapy. These data strongly support the notion that PMC-403 can effectively treat tumors even with low doses of radiation by stabilizing tumor blood vessels.

Citation Format: Eun-Ah Lee, Beom Yong Park, Nuri Kang, Cheon Ho Park, Weon Sup Lee. Tumor vessel normalization by a novel anti-TIE2 antibody PMC-403 enhances the effectiveness of radiation therapy on cancer therapy [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 5970.
Abstract 5978: Real time pathologic diagnosis of digital biopsy during lung cancer surgery using confocal laser endomicroscopy
Authors: Kim Tae-Jung , Sa Young Jo , Kim Sung Ah , Hwang Kyungmin
Published: June 2022

Purpose: Portable hand held confocal laser endomicroscopy is a novel, noninvasive technology that provides real time imaging during biopsy procedure and could be used to perform rapid histopathological diagnosis. However, its ability of determining whether the ex vivo samples obtained during lung cancer surgery or biopsy are tumor or non tumor is yet to be evaluated. Methods: Real time imaging of 75 surgical samples was performed. In preanalytic process, the samples were stained with fluorescent dye. Benign and malignant histologic structures were classified by cell density, nuclear size variability, and nuclear arrangement, which are compared with HE images from the same section.

Results: We defined the confocal laser endomicroscopy atypia classification from the findings of the cells and made a comprehensive atlas of digital biopsy images for pleural surface, bronchus, and lung parenchyma. A total of 3,734 digital images were correlated with 75 frozen lung specimens. Three pathologists assessed digital biopsy images for defining cancer versus non cancer. Overall accuracy was 92.2%, The sensitivity for malignancy was 100% and the specificity was 84.4%. Both inter observer (k = 0.51) and intra observer (k = 0.52) agreement confirmed moderate agreement. Conclusion: Portable confocal laser endomicroscopy was useful to differentiate malignant from benign, to assess the histology of pleural surface and bronchial margin and might be substitute for rapid on site evaluation of pathologic diagnosis during surgical or biopsy procedure.

Citation Format: Tae-Jung Kim, Young Jo Sa, Sung Ah Kim, Kyungmin Hwang. Real time pathologic diagnosis of digital biopsy during lung cancer surgery using confocal laser endomicroscopy [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 5978.
Abstract 5983: Gene profile analysis of breast cancer lung metastasis, liver metastasis, brain metastasis and primary tumor
Authors: Fang Yanman , Zhang Hushan , Zou Jiali , Zhou Fulin , Tang Xuejie , Yang Yi , Luo Li , Ding Yu
Published: June 2022

Background: The incidence of male breast cancer (MBC) is lower than female breast cancer (FBC), representing 1% of all breast cancer worldwide. The difference with FBC may affect the treatment strategy of MBC, but the necessary precondition is that we should know more about the difference between FBC and MBC. However, Because of its low incidence that the relevant data about MBC is relatively scarce. Here we report some data about gene mutation of MBC, and compared it with FBC.

Methods: Blood or tumor Tissue samples of breast cancer were collected and subjected to NGS in a College of American Pathologists (CAP)-certified and Clinical Laboratory Improvement Amendments (CLIA)-accredited lab for gene mutation analysis (3D Medicines Inc.,Shanghai, China). Statistical analysis was performed using GraphPad Prism (version 7.01) and SPSS version 21.0 (SPSS,Inc.).

Results: Approximately 182 samples were analyzed, among which, exist 82 samples were from primary tissue, 66 samples were breast cancer liver metastases, 20 were lung metastases and 14 were brain metastases. High genomic heterogeneity was found among primary breast cancer tissue, breast cancer liver metastases, lung metastases, brain metastases, that is at least 20 gene mutations were found in these BC samples, while different gene mutations were showed among primary and different metastases tissues. The top five mutated gene in primary tumor tissue were TP53(80.49%), PIK3CA(53.66%), FGF19(30.49%), ERBB2(26.83%), MYC(23.17%); in liver metastases tissues were TP53(54.55%), FGF19(39.39%), PIK3CA(37.88%), ERBB2(21.21%), CCND1(19.70%); in lung metastases tissues were TP53(70.00%), PIK3CA(55.00%), ERBB2,MCL1(25.00%), CDKN2A/2B(20.00%), CCND1, NF1, CDK12(15.00%), CCND2, MYC, PTEN, GNAS, FGF19, PTK2, FGFR1(10.00%); in brain metastases tissues were TP53(42.86%), PIK3CA(28.57%), ZNF703, PREX2, JAK3, MYC, FGFR1(14.29%). Besides, levels of TMB were also different among primary tissue, liver, lung and brain metastases tissues. Higher TMB level were showed in liver and brain metastases tissue than primary and lung metastases tissues (p<0.001).

Conclusions: Different metastases of breast cancer were different in gene mutation landscape, this imply these patients should be managed differently.

Citation Format: Yanman Fang, Hushan Zhang, Jiali Zou, Fulin Zhou, Xuejie Tang, Yi Yang, Li Luo, Yu Ding. Gene profile analysis of breast cancer lung metastasis, liver metastasis, brain metastasis and primary tumor [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 5983.
Abstract 5986: WAVE2/EGF signaling and miR29 regulates WAVE2 to promote tumor growth and metastasis in triple negative breast cancer
Authors: Rana Priyanka S. , Wang Wei , Alkrekshi Akram , Markovic Vesna , Sossey-Alaoui Khalid
Published: June 2022

Breast cancer is the most frequently diagnosed malignancy in women and is one of the leading causes of death due to cancer invasion, metastasis, and resistance to therapies. Among its variants, triple-negative breast cancer (TNBC) is considered the most aggressive due to its early age of diagnosis, invasive and metastatic properties with poor prognosis. TNBC is a type of breast cancer that lacks expression of the receptors that are commonly found in this type of cancer (estrogen, progesterone, and HER2), which makes it a more difficult target for treatment. Cancer invasiveness and metastasis are known to be promoted by increased cell motility and upregulation of the Wiskott-Aldrich syndrome protein (WASP) and WASP family verprolin-homologous protein (WAVE) - WAVE1, WAVE2, and WAVE3. While the contribution of WAVE2 to cancer progression is well documented, the WAVE2-mediated regulation of TNBC oncogenic properties is still under investigated. In this study, we show that WAVE2 plays a significant role in TNBC development, progression, and metastasis. To investigate this notion, we used CRISPR/Cas9 gene editing to successfully inhibit WAVE2 expression in several TNBC cell lines. In vitro, loss of WAVE2 expression inhibited 3D tumorsphere formation and invasion of TNBC cells. In vivo, in mouse models for TNBC, loss of WAVE2 inhibited tumor growth and metastasis. Mechanistically, we show that the WAVE2-mediated modulation of the oncogenic behavior of TNBC cells is in part due to its phosphorylation downstream of EGF. At the post transcriptional level, we found WAVE2 expression to be regulated by miR29. Together, our findings identify a novel post-translational WAVE2/EGF oncogenic signaling axis, along with a post-transcriptional regulatory axis downstream of miR29, which together regulate the oncogenic activity of WAVE2 in TNBC and open a new therapeutic strategy for the treatment of TNBC.

Citation Format: Priyanka S. Rana, Wei Wang, Akram Alkrekshi, Vesna Markovic, Khalid Sossey-Alaoui. WAVE2/EGF signaling and miR29 regulates WAVE2 to promote tumor growth and metastasis in triple negative breast 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 5986.
Abstract 5989: Brain metastasis mouse models for the evaluation of multikine inhibitors on ROS1-fusion-positive lung cancer
Authors: Qi Hui , Xiong Xia , Liu Huanhuan , Yang Yuying , Dai Chunlei , Zhao Haoxia , Han Bingrui , Tang Xuzhen , Ji Qunsheng
Published: June 2022

Lung cancer is the leading cause of cancer-related death worldwide. Nearly 80% of lung cancers are non-small cell lung cancer (NSCLC) and 60% of them are diagnosed at the metastatic stage. Brain metastases affect more than 20% of NSCLC patients with poor prognosis and disabling symptoms. However, few therapies have been approved for the treatment of lung cancer brain metastases. A panel of rapid, predictive and clinically-relevant animal models are urgently needed to study the biology of brain metastases and to identify effective therapeutic approaches.Considering both progression and treatment of lung cancer brain metastases will be limited by the blood-brain barrier (BBB), here we describe methods for efficient establishment of brain metastases mouse models via different injection route (intracranial, intracardiac or intracarotid), to identify a model that permits tumor cell growth in the unique brain microenvironment without compromising BBB. Meanwhile, we established a novel ROS1 positive patient-derived xenograft (PDX) model together with its PDC sub-line, in which the exon 2 of SDC4 was fused to the exon 32 of ROS1 (SDC4ex2-ROS1ex32). The cell has been transduced with firefly luciferase expression vector for in vivo imaging detection. Three ROS1 inhibitors were tested on these brain metastasis and subcutaneous models to compare their efficacies. In summary, we established brain metastases lung cancer PDX models as reliable platforms for interrogation of targets and kinetics related mechanism and for drug screening. The use of such techniques will increase our knowledge of the metastatic process and help identify new targets of cancer metastasis.

Citation Format: Hui Qi, Xia Xiong, Huanhuan Liu, Yuying Yang, Chunlei Dai, Haoxia Zhao, Bingrui Han, Xuzhen Tang, Qunsheng Ji. Brain metastasis mouse models for the evaluation of multikine inhibitors on ROS1-fusion-positive 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 5989.
Abstract 5996: Single tube PCR-based NGS assay for detection of multiple gene fusions from cell free total nucleic acid
Authors: Modak Tejashree , Li Jilong , Polvino Sean , Ke Yue , Wang Zhaohui , Kotlar Alex V.
Published: June 2022

Introduction: Recently, liquid biopsy has begun to be adopted for treatment selection and drug-resistance assessment for cancers. However, currently liquid biopsy is mostly focused on analyzing cell-free DNA (cfDNA). Though analyzing RNA provides more accurate analysis for functional gene fusions, little is known about the stability and availability of cfRNA (cell free RNA). Here we report our one-tube robust cfTNA (cell free total nucleic acids) assay to analyze ctDNA and ctRNA simultaneously.

Methods: We assessed Pillar’s cell free TNA panel performance on 2 confirmed fusion positive and 5 fusion negative samples that were serially diluted to obtain a total of 88 samples including replicates. An input of 10ng cfTNA was used for fusion positive samples with fusion frequency ranging from 0.2% to 20%. A range of 5 to 30ng input was used for fusion negative samples. The amount of input cfRNA was estimated as equal to the cfDNA input. Total nucleic acids were extracted and cfRNA was reverse transcribed. Amplicons targeted the specific breakpoints of 36 known fusion transcripts. Fusion positive samples were diluted for library prep to achieve fusion frequency of 0.5%, 1%, 4%, 5% and 20%. Novaseq was used for sequencing with an average depth of 200,000X and downsampled by 25%. Pillar Variant Analysis Toolkit (PiVAT®) was used to map and call fusions.

Results: Among dilutions of >=4%, PiVAT achieved >=99% sensitivity and >=99.9% specificity in calling RNA fusions from 10ng input. Even at extremely low dilutions of <=1%, PiVAT achieved >= 50% sensitivity.

Conclusion: Pillar’s cell free TNA panel offers a simple, rapid, and reliable workflow that enables detection of a broad range of clinically relevant gene fusions. Detection of fusion events in cell-free samples from noninvasive liquid biopsies represents an important step forward in cancer diagnosis and monitoring.

Citation Format: Tejashree Modak, Jilong Li, Sean Polvino, Yue Ke, Zhaohui Wang, Alex V. Kotlar. Single tube PCR-based NGS assay for detection of multiple gene fusions from cell free total nucleic acid [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 5996.
Abstract 5997: The role of neutrophil extracellular trap-CCDC25-integrin linked kinase complex in pancreatic cancer progression and metastasis
Authors: McDonald Paul C. , Brown Wells S. , Gerbec Zack , Awrey Shannon , Karasinska Joanna , Schaeffer David , Renouf Daniel , Stanger Ben , Dedhar Shoukat
Published: June 2022

Neutrophils and neutrophil extracellular traps (NETs) contribute to the hypoxic, immunosuppressive pancreatic ductal adenocarcinoma (PDAC) tumour microenvironment (TME) which promotes treatment resistance, increased invasion and metastasis. While recent studies have demonstrated a critical role of tumour cell CCDC25-ILK complex in promoting NET-DNA-induced tumour metastasis in breast cancer, here we have investigated the presence and function of NETosis-induced CCDC25-ILK signaling in the context of pancreatic cancer. We demonstrate that CCDC25 and ILK are expressed in human PDAC cell lines, patient-derived xenograft (PDX) cell lines and in cell lines derived from the KPCY genetic model of PDAC. Importantly, co-immunoprecipitation analyses showed that endogenous CCDC25 and ILK interact in PDAC cells, and the addition of neutrophil-derived NET-DNA resulted in a greater amount of ILK associated with CCDC25 and in increased wound-induced migration. Time-lapse imaging demonstrated a dramatic effect of inhibition of ILK activity on NET-DNA-induced migration by tumour cells in vitro. In vivo, co-localization of CCDC25 and ILK was observed in tumours and liver metastases from both the KPCY GEMM and the MIA PaCa-2 human PDAC xenograft models that were stained by multispectral immunofluorescence for these markers and analyzed by confocal microscopy. Similarly, both tumours and liver metastases showed the presence of infiltrating neutrophils and NETs as indicated by staining for myeloperoxidase and citrullinated histone H3. Collectively, these data identify the neutrophil extracellular trap-induced CCDC25-ILK interaction as an important contributor to PDAC progression and suggest the potential for targeting this axis to prevent NET-induced PDAC progression and metastasis.

Citation Format: Paul C. McDonald, Wells S. Brown, Zack Gerbec, Shannon Awrey, Joanna Karasinska, David Schaeffer, Daniel Renouf, Ben Stanger, Shoukat Dedhar. The role of neutrophil extracellular trap-CCDC25-integrin linked kinase complex in pancreatic cancer progression and metastasis [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 5997.
Abstract 6063: FOXK2 promotes stemness in ovarian cancer by regulating unfolded protein response signaling
Authors: Zhang Yaqi , Wang Yinu , Zhao Guangyuan , Tanner Edward J. , Adli Mazhar , Matei Daniela E.
Published: June 2022

Introduction: Cancer stem cells (CSCs), a rare cell population within tumors that possesses self-renewal and tumor initiation capacity, have been implicated in cancer relapse and resistance to chemotherapy. In ovarian cancer (OC), CSCs are characterized by a high expression of aldehyde dehydrogenase (ALDH). Understanding the regulatory programs promoting stemness could identify new treatments. We identified FOXK2 as a key transcription factor (TF) that promotes stemness in OC.

Methods: We generated FOXK2 knockdown OC models by stable transduction of shRNA targeting this TF and measured the population of ALDH+ CSCs, expression of stemness-associated genes, and in vivo tumor initiation capacities. Transcriptomic and ChIP-seq analyses were used to identify key FOXK2 targets. CRISPR-dCas9 genome editing was used to verify the regulatory function of FOXK2. Rescue experiments validated the roles of a newly discovered target gene of FOXK2.

Results: Through gene expression analyses in CSCs vs. non-CSCs, we identified FOXK2 as a highly expressed stemness-specific TF in OC. Genetic depletion of FOXK2 diminished stemness features and reduced in vivo tumor initiation capacity. Through ChIP-sequencing we discovered that FOXK2 directly regulates IRE1α (ERN1 gene) expression, a key sensor for the unfolded protein response (UPR). ChIP-sequencing analysis further revealed that FOXK2 binds to an intronic regulatory element of ERN1. Blocking FOXK2 from binding to this enhancer by using a CRISPR/dCas9 diminished IRE1α transcription. At the molecular level, FOXK2-driven upregulation of IRE1α led to alternative XBP1 mRNA splicing and activation of stemness pathways, while genetic or pharmacological blockade of IRE1α inhibited CSCs in OC models.

Conclusion: Our data establish a new function for FOXK2 as a key transcriptional regulator of CSCs and a mediator of the UPR, providing insight into potentially targetable new pathways in ovarian CSCs.

Citation Format: Yaqi Zhang, Yinu Wang, Guangyuan Zhao, Edward J. Tanner, Mazhar Adli, Daniela E. Matei. FOXK2 promotes stemness in ovarian cancer by regulating unfolded protein response signaling [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 6063.
Abstract 6069: HOXA9 overexpression contributes to stem cell overpopulation that drives development and growth of colorectal cancer
Authors: Osmond Brian T. , Facey Caroline O. , Opdenaker Lynn M. , Boman Bruce M.
Published: June 2022

HOX genes encode master regulatory transcription factors that regulate stem cells (SCs), but their role in carcinogenesis is not fully understood. SC overpopulation drives colorectal cancer (CRC) development, which is also linked with the aberrant expression of specific HOX genes. One such gene is HOXA9. Our goal is to find out how HOXA9-regulatory mechanisms play a role in the SC origin of CRC. Previous research in our lab reported that HOXA9 as well as other HOX proteins (e.g. HOXA4 & HOXD10) are selectively expressed in human colonic cancer SCs (CSCs). The SC marker aldehyde dehydrogenase (ALDH) and other retinoic acid (RA) pathway components are also selectively expressed in CSCs. Moreover, HOXA9 expression is positively correlated with expression of ALDH genes such as ALDH1A1. These findings suggest that RA signaling regulates HOXA9 gene expression in colonic ALDH+ SCs.

Hypothesis: Dysregulated HOXA9 expression leads to ALDH+ SC overpopulation, which contributes to CRC growth and development. We used immunofluorescent (IF) mapping to determine the histologic location of HOXA9+ cells in relation to colonic SCs and other differentiated colonic cell types in normal and neoplastic human colonic tissues. We also measured the effects of all-trans retinoic acid (ATRA) and the ALDH enzyme inhibitor diethylamino benzaldehyde (DEAB) on HOXA9 expression, cell proliferation and differentiation using CRC cell lines. Immunofluorescence mapping indicated that not only is HOXA9 selectively expressed in ALDH-positive SCs, but that HOXA9 expression is increased in colon carcinoma tissue compared to normal colon epithelium. Our results also showed that both ATRA and DEAB treatment decreased the proliferation of HT29 and SW480 CRC cell lines. Western blot analysis showed that HOXA9 has reduced expression in ATRA-treated HT29 CRC cells. Additionally, our independent bioinformatics analyses show that HOXA9 is predominantly expressed in both stem cells and neuroendocrine cells (NECs) in normal human colonic epithelium. Taken together, these findings indicate that regulation of HOXA9 gene expression by RA signaling controls differentiation of ALDH+ SCs along the NEC lineage in human colon tissue. Thus, our results show that HOXA9 is regulated by RA signaling and when dysregulated, contributes to CRC development and growth. These findings provide a mechanism that may be targeted in the design of novel treatments against SC overpopulation in CRC.

Citation Format: Brian T. Osmond, Caroline O. Facey, Lynn M. Opdenaker, Bruce M. Boman. HOXA9 overexpression contributes to stem cell overpopulation that drives development and growth of colorectal 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 6069.
Abstract 6071: Activation of NOTCH signaling via DLL1 is mediated by APE1-redox-dependent NF-κB activation in esophageal adenocarcinoma
Authors: Chen Lei , Lu Heng , Peng Dunfa , Cao Longlong , Chen Zheng , Bhat Ajaz , Zaika Alexander , Zhang Shutian , El-Rifai Wael
Published: June 2022

Esophageal adenocarcinoma (EAC) is one of the most lethal of all human malignancies. EAC arises in the setting of Barrett’s esophagus, which is an intestinal metaplastic precursor lesion that develops from chronic reflux of gastrointestinal contents (especially acidic bile salts). Here we report that NOTCH signaling is activated in the esophagus through a unique pathway during exposure to acidic bile salts and progression to EAC. Using a combination of public databases, EAC cell line models, transgenic mice, and patient tissue samples, we detected significant upregulation of several NOTCH signaling components in EAC. Activated NOTCH signaling was confirmed by nuclear accumulation of NOTCH1 cleaved fragment (NICD) with corresponding up-regulation of NOTCH targets in EAC cells in response to acidic bile salts. Moreover, we identified DLL1 as the predominant ligand contributing NOTCH1 activation. Remarkably, DLL1 was regulated by direct cross talk between redox and inflammatory pathways that are activated during both reflux and malignant transformation. Mechanistically, the APE1 redox function transcriptionally up-regulated NF-κB in response to bile salts. This licensed NF-κB to transcriptionally up-regulate DLL1 to activate and stimulate downstream NOTCH1 signaling, thereby defining a novel APE1-NF-kB-NOTCH pro-tumorigenic pathway. This pathway was important for maintaining tumor initiating (cancer “stem cell-like”) properties in vitro, recurrently detected in genetically engineered mouse models of EAC and in EAC patient samples in vivo, and portended an overall poor prognosis. Collectively, these findings indicate that progression from chronic injury to malignancy in the esophagus is driven by a unique mechanism that links redox balance, inflammation, embryonic development (NOTCH) together into a common pro-tumorigenic pathway that is intrinsic to EAC cells.

Citation Format: Lei Chen, Heng Lu, Dunfa Peng, Longlong Cao, Zheng Chen, Ajaz Bhat, Alexander Zaika, Shutian Zhang, Wael El-Rifai. Activation of NOTCH signaling via DLL1 is mediated by APE1-redox-dependent NF-κB activation in esophageal adenocarcinoma [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 6071.
Abstract 6075: ROS-Nrf2-ABCB1 axis accompanied with pFyn nuclear accumulation plays pivotal roles in vinorelbine resistance in non-small cell lung cancer
Authors: Tamari Shigeyuki , Menju Toshi , Toyazaki Toshiya , Miyamoto Hideaki , Chiba Naohisa , Noguchi Misa , Date Hiroshi
Published: June 2022

Introduction: Adjuvant cisplatin-vinorelbine is standard therapy for stage II/III lung cancer. Poor survival in lung cancer can be attributed to vinorelbine resistance arising from ABC sub-family B member 1 (ABCB1) and phosphorylated Fyn (pFyn) overexpression. However, the underlying mechanism remains unclear. Nrf2 regulates the cell-protective ABC family and activates nuclear transport of Fyn. Here, we aimed to evaluate the ROS-Nrf2-ABCB1 axis in both vinorelbine-resistant (VR) cells and clinical samples.

Methods: We established VR cells by exposing H1299 cells to vinorelbine, and determined intracellular reactive oxygen species (ROS) level of H1299 cells induced by vinorelbine using DCFH-DA. We evaluated the total and subcellular expression of Nrf2, ABCB1, and pFyn in VR cells and performed immunofluorescence of VR cells to confirm the precise subcellular localization of pFyn. Cell viability assay was used to examine whether the sensitivity of VR cells to vinorelbine depends on Nrf2 activity. For immunohistochemistry, we used 104 tissue samples from lung cancer patients who underwent surgery followed by cisplatin-vinorelbine treatment between December 2006 and June 2018.

Results: Persistent exposure to vinorelbine induced intracellular ROS formation of H1299 cells. Both ABCB1 and Nrf2 were overexpressed in VR cells, and ABCB1 expression was dependent on Nrf2 downstream activation. Subcellular fractionation and immunofluorescence revealed nuclear accumulation of pFyn in VR cells. Decreased expression of Nrf2 restored the sensitivity of VR cells to vinorelbine. In the 104 surgical samples, Nrf2 and ABCB1 were significantly associated with disease-free survival (p = 0.029 and 0.035, respectively), and pFyn was associated with overall survival (p = 0.040).

Conclusions: ROS-Nrf2 upregulates ABCB1 and accompanied by nuclear accumulation of pFyn, induces vinorelbine resistance. Our findings may facilitate drug resistance prevention strategies or new drug target development against non-small cell lung cancer.

Citation Format: Shigeyuki Tamari, Toshi Menju, Toshiya Toyazaki, Hideaki Miyamoto, Naohisa Chiba, Misa Noguchi, Hiroshi Date. ROS-Nrf2-ABCB1 axis accompanied with pFyn nuclear accumulation plays pivotal roles in vinorelbine resistance in non-small cell 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 6075.
Abstract 6081: The transcription factor SPDEF promotes the survival of mucinous pancreatic ductal adenocarcinoma of the classical subtype
Authors: Tonelli Claudia , Yordanov Georgi N. , Hao Yuan , Deschênes Astrid , Brosnan Erin , Doshi Abishek , Park Youngkyu , Preall Jonathan , Tuveson David A.
Published: June 2022

Pancreatic ductal adenocarcinoma (PDA) is characterized by intra-tumoral heterogeneity. Using single-cell RNA sequencing, we reveal multiple tumor subpopulations distinguished by their differentiation state and associated with different stages of tumor progression. We identify SPDEF as a factor required for tumorigenesis in pancreatic cancer cells of epithelial and mucinous nature. SPDEF levels correlate with glandular differentiation and high secretory activity, which are characteristic features of human PDA of the classical subtype. SPDEF can drive mucus production in pancreatic cancer cells and is essential for the growth of mucus-secreting tumors by regulating endoplasmic reticulum (ER) activity. These findings offer insights into the factors controlling differentiation in PDA and may inform new therapeutic strategies with improved efficacy.

Citation Format: Claudia Tonelli, Georgi N. Yordanov, Yuan Hao, Astrid Deschênes, Erin Brosnan, Abishek Doshi, Youngkyu Park, Jonathan Preall, David A. Tuveson. The transcription factor SPDEF promotes the survival of mucinous pancreatic ductal adenocarcinoma of the classical subtype [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 6081.
Abstract 6094: SRC inhibitor saracatinib enhances efficacy of PD-1/PD-L1 immune checkpoint blockade in non-small cell lung cancer
Authors: Lu Fei-Teng , Luo Fan , Qiu Miao-Zhen , Cao Jia-Xin , Luo Qiu-Yun , Yang Da-Jun , Zhao Hong-Yun
Published: June 2022

Background: Although immune checkpoint inhibitors (ICI) included PD-1/PD-L1 have demonstrated durable therapeutic responses in some cases, a significant portion of patients with cancer remain nonresponsive (e.g. only ~20% with anti-PD-1 in lung cancer). Substantial limitations in the rate of response underscore the need for suitable combination therapies. SRC family kinases is overexpressed or activated in multiple human malignancies, as a candidate target in regulating antitumor immunity. Saracatinib, an orally bioavailable multi-kinase inhibitor, has demonstrated clinical efficacy in CML. Here, we proved saracatinib as SRC inhibitor that were capable of increasing the antitumor immunity in non-small lung cancer.

Method: Cytotoxicity assays were used to determined antitumor activities of saracatinib in non-small lung cancer cell lines. The expression of PD-L1 expression in saracatinib 1 treated cell lines was investigated by western blots, quantitative PCR and flow cytometry. The effect of saracatinib in antitumor immunity alone or in combination with immunotherapy was also determined in vitro and in vivo. RNAi, overexpression, and gene expression analyses were used to define underlying mechanisms.

Result: We found that SRC inhibitor saracatinib robustly decreased expression of PD-L1 in non-small lung cancer cells and animal models. The observed inhibitory effect was time- and concentration-dependent. Furthermore, our results showed that saracatinib and anti-PD-L1 combined to improve T-cell-mediated killing of tumor cells in vitro and in vivo, which were associated with activation of CD8+ T cytotoxic cell included expression of IFN-γ and granzyme-B. Prolonged survival was also observed in mice treated with saracatinib in combination with PD-L1 blockade.

Conclusion: Our in vivo and in vitro data demonstrated that SRC inhibitor saracatinib enhance T-cell-mediated anti-tumor immune responses in non-small lung cancer. We also provide evidence that support the combination of saracatinib and anti-PD-1/PD-L1 as a potential combination therapeutic approach to increase the efficacy of immunotherapy in non-small lung cancer.

Citation Format: Fei-Teng Lu, Fan Luo, Miao-Zhen Qiu, Jia-Xin Cao, Qiu-Yun Luo, Da-Jun Yang, Hong-Yun Zhao. SRC inhibitor saracatinib enhances efficacy of PD-1/PD-L1 immune checkpoint blockade in non-small cell 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 6094.