bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3062: ROMO1 mediates mitochondrial function links to neuroendocrine differentiation of prostate cancer enhancement by the nuclear PKLR/MYCN/MAX complex
Authors: Liu Yen-Nien , Chen Wei-Yu , Dung Phan Vu Thuy , Yeh Hsiu-Lien , Chen Zi-Qing , Wang Wan-Hsin , Wen Yu-Ching
Published: April 2023

Conventional treatment of prostate cancer (PCa) uses androgen deprivation therapy (ADT) to inhibit androgen receptor (AR) signaling-driven tumor progression. ADT-induced PCa recurrence may progress to AR-negative phenotype with neuroendocrine (NE) features on histology, which is associated with metabolic disturbances and poor prognosis. However, the association between the metabolic pathways that regulate mitochondrial function and promote NE differentiation (NED) in PCa remains unclear. Here, we show a regulatory mechanism in PCa pathogenesis whereby overexpression of pyruvate kinase L/R (PKLR) mediates mitochondrial disorder and oxidative stress through upregulation of reactive oxygen species modulator 1 (ROMO1), thereby promoting NED-related aggressiveness. ADT mediates the nuclear translocation of PKLR, which binds to the MYCN/MAX complex to upregulate ROMO1 and NE-related genes, leading to mitochondrial dysfunction and NED in PCa. Targeting nuclear PKLR/MYCN using bromodomain and extra-terminal motif (BET) inhibitors has the potential to reduce PKLR/MYCN-driven NED. Abundant ROMO1 in serum samples may provide prognostic information in patients with ADT-induced NED. Our findings link ADT resistance-driven metabolic reprogramming to NED of PCa through upregulation of PKLR/MYCN/ROMO1 signaling, and further explored the clinical significance of targeting the nuclear PKLR/MYCN complex and evaluated serum ROMO1 as a potential biomarker for aggressive NE-like PCa.

Citation Format: Yen-Nien Liu, Wei-Yu Chen, Phan Vu Thuy Dung, Hsiu-Lien Yeh, Zi-Qing Chen, Wan-Hsin Wang, Yu-Ching Wen. ROMO1 mediates mitochondrial function links to neuroendocrine differentiation of prostate cancer enhancement by the nuclear PKLR/MYCN/MAX complex [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3062.
Abstract 3230: Tumor microenvironment modulation by immunotherapy sensitizes solid tumors to radiation
Authors: Kang Minjeong , Lee DaeYong , Wang Yifan , Kim Betty , Jiang Wen
Published: April 2023

Radiation therapy (RT) has been prevalently implemented to treat cancer in clinical settings by inducing DNA damage in tumor cells. However, tumor microenvironmental factors such as severe hypoxic conditions and vascular anomalies attenuate the therapeutic efficacy of RT, thus resulting in unsatisfactory outcomes in solid tumors. Recently, Immune checkpoint blockade (ICB) has improved immune recognition to tumor cells and normalized tumor vasculature to diminish hypoxia in solid tumors. Herein, we suggest that RT combined with ICB synergistically alters tumor microenvironments, thus facilitating the recruitment of cytotoxic immune cells into tumors as well as sensitizing radiation effectiveness. RT in combination with ICB not only normalized tumor vasculature to reduce hypoxia, but also increased anti-tumor effects from immune-mediated toxicity and DNA damage-mediated apoptosis in several solid tumor-bearing mouse models to a greater extent than did either monotreatment of RT or ICB. This study will provide mechanistic insight into synergistic effects conferred by RT and ICB as well as a strong preclinical rationale for combining the two therapeutic modalities in human cancers.

Citation Format: Minjeong Kang, DaeYong Lee, Yifan Wang, Betty Kim, Wen Jiang. Tumor microenvironment modulation by immunotherapy sensitizes solid tumors to radiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3230.
Abstract 3249: Circulating cytokine associations with clinical outcomes in melanoma patients treated with checkpoint inhibitors
Authors: Chen Jiajia , Tarantino Giuseppe , Severgnini Mariano , Baginska Joanna , Giobbie-Hurder Anita , Manos Michael , Russell Janice D. , Weirather Jason L. , Pfaff Kathleen , Huang Amy Y. , Rodig Scott J. , Ranashinge Srinika , Liu David , Hodi F.Stephen
Published: April 2023

Background: aCTLA-4/aPD-1 combination therapy has significantly improved clinical outcomes in patients with metastatic melanoma, with 50%-60% of patients responding to treatment, but predictors of response are poorly characterized. In this study, we set out to evaluate the hypothesis that circulating cytokines and peripheral white blood cell composition may predict response to therapy.

Methods: We quantified cytokines and complete blood counts from 89 patients with advanced melanoma treated with combination therapy from three points in time: pre-treatment, one month and approximately three months after starting therapy. We evaluated the association between peripheral blood circulating features, cytokines and clinical labs values, and clinical outcome using predictive modeling, and investigated the prognostic values of these features using survival analysis.

Results: Pre-treatment, patients with durable clinical benefit (DCB) had higher IL23, lower CXCL6, and lower IL10 levels. The ratio of neutrophils to lymphocytes (NLR) pre-treatment was not associated with response, but higher NLR one month after starting therapy predicted poorer progression free survival (PFS) and overall survival (OS), primarily driven by an increase in absolute lymphocytes. A multivariate model demonstrated that baseline CXCL6, IL10, IL23 were independent predictors of therapy response, and the combined model has reached an area under the curve (AUC) of 0.8 in prediction of response to combination therapy. Low IL10 and CXCL6 identified an excellent prognosis group, while high CXCL6 and low IL23 characterized a very poor prognosis group of patients.

Conclusions: Baseline measurement of cytokines IL23, CXCL6, and IL10 can predict metastatic melanoma patients’ response to ipi/nivo combination treatment. Higher neutrophils-to-lymphocytes ratio approximately one month after therapy started associates with worse survival outcome. Our study presented circulating features from a relatively accessible and less costly avenue, peripheral blood, to be predictive of response to combination immune checkpoint blockade.

Citation Format: Jiajia Chen, Giuseppe Tarantino, Mariano Severgnini, Joanna Baginska, Anita Giobbie-Hurder, Michael Manos, Janice D. Russell, Jason L. Weirather, Kathleen Pfaff, Amy Y. Huang, Scott J. Rodig, Srinika Ranashinge, David Liu, F.Stephen Hodi. Circulating cytokine associations with clinical outcomes in melanoma patients treated with checkpoint inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3249.
Abstract 326: Identification of novel substrates of the UBR5 E3 ubiquitin ligase in ovarian cancer
Authors: Eblen Scott T. , Bethard Jennifer R. , Rogers D Ralph , Daoud Abdelkader , Madan Lalima K. , Ball Lauren E.
Published: April 2023

The ubiquitin proteasome system regulates the degradation of proteins in order to maintain cellular homeostasis, which is often disrupted in cancer cells. The UBR5 (Ubiquitin Protein Ligase E3 Component N-Recognin 5) protein is an E3 ubiquitin ligase that catalyzes the direct ubiquitination of protein substrates, most often resulting in substrate protein degradation. UBR5 is the human homolog of the Drosophilia hyperplastic discs (hyd) tumor suppressor gene. However, ubr5 is amplified and over-expressed in human ovarian and breast cancers, among others, and thought to play more of a tumor-promoting role in cancer. We previously demonstrated that targeting UBR5 expression in human ovarian cancer xenografts in mice with liposomal delivery of UBR5 siRNA inhibited tumor growth and sensitized tumors to the DNA-damaging agent cisplatin. To identify novel substrates of UBR5-stimulated degradation, we transduced OVCAR5 cells with doxycycline-inducible shRNA targeting UBR5 or a control shRNA and metabolically labelled proteins with heavy or light lysine and arginine using Stable Isotope Labeling with Amino acids in Cell culture (SILAC). Doxycycline was added for the last 48 hr and cisplatin for the last 12 hrs before cell harvest. Equal amounts of protein from both cultures were mixed, digested with trypsin and analyzed by LC-MS/MS (Orbitrap Elite MS) in three independent experiments including a label swap control. Peptides and proteins were identified and quantified using MaxQuant (FDR<0.01). To identify proteins differentially expressed after UBR5 knockdown, relative protein abundance was compared by two-sided t-test with permutation-based correction of the p-value. We identified multiple proteins with increased expression following UBR5 knockdown, suggesting these proteins are regulated by ubiquitin-mediated degradation by UBR5. We validated UBR5-mediated regulation of several proteins by Western blotting, including an oncogene involved in breast and ovarian cancer progression, which we further characterized. In summary, we were able to identify several novel UBR5 candidate substrates whose expression is increased following UBR5 knockdown. Further characterization of these substrates will allow us to identify novel mechanisms of UBR5 regulation of tumor progression.

Citation Format: Scott T. Eblen, Jennifer R. Bethard, D Ralph Rogers, Abdelkader Daoud, Lalima K. Madan, Lauren E. Ball. Identification of novel substrates of the UBR5 E3 ubiquitin ligase in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 326.
Abstract 3304: Immune repertoire sequencing enables accurate clonality determination
Authors: Song Chen , Erijman Ariel , Lund Sean , Langhorst Bradley W. , Liu Pingfang
Published: April 2023

The study of complex immunological diseases and tumor microenvironments has advanced through recent developments in sequencing of the immune repertoire. Using this approach, the interrogation of disease progression is facilitated through analysis of millions of V(D)J combinations from B cell receptors (BCRs) and T cell receptors (TCRs). One major challenge of immune repertoire sequencing is to accurately capture the structural and sequence complexities of antibodies and TCR genes during both library preparation and bioinformatic analysis. In addition, the lack of standard reference controls makes assessing sequencing accuracy and sensitivity challenging. Here, we present a method for accurate sequencing of full-length immune gene repertoires of B cells and T cells.

RNA extracted from peripheral blood mononuclear cells (PBMCs) or tissues were used for reverse transcription, during which unique molecular identifiers (UMIs) were added to discretely barcode each mRNA molecule. BCR- and TCR-specific PCR primers were used to enrich full-length BCR and TCR sequences. We have implemented a data analysis pipeline to assemble the full length BCR/TCR transcripts and to collapse PCR copies of each mRNA fragment into a single consensus sequence using UMIs. UMI incorporation enables the absolute quantification of input RNA molecules and accurate ranking of antibody/TCR clone abundance. Furthermore, we developed an RNA control pool that contains a large dynamic range of BCR and TCR sequences with a variety of V(D)J combinations, which enables sensitivity and accuracy evaluation of BCR and TCR sequencing.

Our immune repertoire sequencing approach allows accurate clonal determination for both BCR and TCR. This technique is applicable for various applications including design of antibody chains for in vitro synthesis, investigation of T cell infiltration of tumor microenvironments, and monitoring of minimal residual disease in cancer patients.

Citation Format: Chen Song, Ariel Erijman, Sean Lund, Bradley W. Langhorst, Pingfang Liu. Immune repertoire sequencing enables accurate clonality determination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3304.
Abstract 3307: A novel targeted RNA sequencing approach for identification of aggressive prostate cancer
Authors: Fredsøe Jacob , Haldrup Christa , Nordentoft Iver , Ulhøi Benedicte , Borre Michael , Sørensen Karina D.
Published: April 2023

Introduction: Prostate cancer (PC) is a highly heterogeneous disease and can be either aggressive, requiring immediate treatment, or remain slow growing and even without treatment present no significant symptoms throughout the man’s life. Currently, there is an unmet need for new tools that can clearly distinguish indolent from aggressive PC in order to guide treatment decisions.PC is a highly heritable disease and so far, approximately 450 genomic regions associated with PC risk have been defined in genome-wide association studies. Most of these regions are however devoid of annotated genes and consequently the underlying mechanisms behind the associations are not easily elucidated. Accordingly, yet uncharacterized genes in PC-associated regions could be involved in PC development/progression.

Methods: We quantified gene expression levels by Kallisto on total RNAseq data from fresh frozen prostate tissue samples from 134 localized PCs, 33 matched adjacent normal (AN) areas as well as 15 PC cell lines. Furthermore, we performed targeted RNAseq covering approximately seven million base pairs divided among 101 PC-risk associated regions (GWAS hits) for in-depth transcriptomic analyses in the same set of samples. HISAT and StringTie were used to identify novel transcripts (genes) with the criteria that a given gene had to have at least 10 reads in more than 50% of all samples to be included. Patient genotyping was performed on Illumina’s global screening array.

Results: In totalRNA seq data we observed significant dysregulation of 10,256 genes (5,685 up- and 4,571 downregulated) between AN and localized PC, including upregulation of know PC-associated genes such as SIM2, PCAT7, TDRD1, and TFF3. Interestingly, we found a correlation between high transcription of TFF3 and high risk of biochemical recurrence (BCR) after radical prostatectomy and furthermore a trans-acting eQTL, the PC-risk associated SNP rs1938781. Here, heterozygote patients with the risk-allele had higher risk of BCR than patients homozygous for the reference allele, indicating a potential functional relationship between the PC risk-SNP and TFF3. Additionally, we detected 207 previously unannotated non-coding genes in the capture RNAseq data, where 148 (71.5%) were undetectable in the matched total RNAseq data indicating the existence of stably albeit lowly expressed genes in the genomic areas containing PC-risk SNPs.

Citation Format: Jacob Fredsøe, Christa Haldrup, Iver Nordentoft, Benedicte Ulhøi, Michael Borre, Karina D. Sørensen. A novel targeted RNA sequencing approach for identification of aggressive prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3307.
Abstract 3308: Predictive biomarkers of response to immune checkpoint inhibitors in metastatic renal cell carcinoma
Authors: Motoshima Takanobu , Miura Yuji , Amami Toshiki , Kitano Shigeyasu , Baba Masaya , Kamba Tomomi
Published: April 2023

Background: Since the introduction of Immune checkpoint inhibitors (ICIs) combination therapy to renal cancer treatment, it has revolutionized the treatment of renal cancer, with many reports of excellent therapeutic effects. However, it is not known which patients will benefit the most.

Materials and methods: PBMC(Peripheral Blood Mononuclear Cells)samples were collected prospectively from patients with advanced and metastatic renal cell carcinoma (RCC) prior to initiation of ICI treatment (baseline) and 2 or 3 weeks after the first cycle of ICI treatment (post-dose 1). We collected PBMCs from 24 patients with a cell count of more than 50,000 were analyzed using antibodies that distinguishCD4 and CD8 cells including Effector, Memory cells population, B cells, Monocyte, NK(Natural Killer) cells, MDSC(Myeloid-Derived Suppressor Cells ), DC(Dendritic cells), CD4 (Th1, Th2, Th17, Tfh), Immune checkpoint molecules including CD223 (LAG3), CD279 (PD1), TIM3, CD152 (CTLA4), CD278 (ICOS), FOXP3.

Results: First, we compared the Fold Change (FC) before and after drug administration in the group that did not respond to treatment (SD: Stable Disease or PD: Progressive disease) and the group that did respond to treatment (PR: Partial Response, CR: Complete Response). The results showed that CD4 positive cells (non-responder Median 0.79 vs responder Median 1.64, p=0.0317, respectively), monocytes: CD56-HLADR+ (non-responder Median 0.87 vs responder Median 1.20, p=0.0159, respectively), but not in other cell populations.Next, we compared the groups that did and did not respond to treatment before and after treatment. We could not identify any biomarker cell population before treatment, the number of CD4 Effector cells (CD4+CD45RO-CD197-) was significantly higher in the post-treatment (PD1) group (PR or CR) than in the non-treatment group (SD or PD). (p=0.0303, respectively)

Conclusion: Elevation of CD4 positive cells and monocytes (CD56-HLADR+) before and after treatment and the number of CD4 Effector cells (CD4+CD45RO-CD197-) after treatment may be biomarkers of treatment response.

Citation Format: Takanobu Motoshima, Yuji Miura, Toshiki Amami, Shigeyasu Kitano, Masaya Baba, Tomomi Kamba. Predictive biomarkers of response to immune checkpoint inhibitors in metastatic renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3308.
Abstract 3352: Development and validation of a serum microRNA-based diagnostic model for early detection of multiple cancers
Authors: Zhou Grace , Hu Hai
Published: April 2023

Introduction: Detecting cancer early improves patients’ survival rate. We previously developed a serum 4-microRNA (miRNA) diagnostic model using lung cancer patient samples, which detected 12 cancer types with high accuracy (Zhang and Hu 2022 Cancers). In this study, we aimed to develop a new diagnostic model based on samples from ovarian cancer patients to evaluate whether model performance depends on the initial cancer type used.

Patients and Methods: Our previous work assembled a serum miRNA microarray dataset from 4 individual datasets from GEO, including 3604 patients across 13 cancer types and 3932 non-cancer controls. In this study, we composed a training set of 191 ovarian cancer patients and 191 non-cancer controls matched by age and gender. Limma analysis was performed to rank differentially expressed miRNAs, and 10-fold cross-validation was used to determine the optimal number of top miRNAs for the final diagnostic model. Model performance was evaluated by area under the curve (AUC) of Receiver Operating Characteristic (ROC) curve analysis, and sensitivity and specificity, using samples not utilized in current or previous model development.

Results: Using this training set, a 10-miRNA model was developed which showed an improved AUC 0.994 as compared to 0.973 from our previous model for ovarian cancer in the testing set. AUC was also substantially improved on sarcoma and breast cancers. With 99.1% specificity, the current model demonstrated at least 70% sensitivity for 10 of the 13 cancer types, comparable to the previous model with 99.3% specificity (Table 1).

Conclusion: Our study showed that blood miRNA-based diagnostic models can be developed for detecting multiple cancers with high accuracy, regardless of initial cancer types used in model development. These results suggested that tumors of different cancers shed common types of miRNAs into the bloodstream, supporting the use of circulating miRNAs as a biomarker for multi-cancer early detection.

ABLE 1. Diagnostic Model Performance Cancer Type N Current Model AUC Current Model Sensitivity Previous Model AUC Previous Model Sensitivity Ovarian 142 0.994 0.80 0.973 0.69 Lung 1358 0.998 0.93 0.999 0.99 Pancreatic 149 0.992 0.76 0.983 0.83 Prostate 40 0.998 0.95 0.996 0.92 Sarcoma 132 0.973 0.52 0.876 0.72 Esophageal 124 0.995 0.90 0.990 0.85 Gastric 150 0.998 0.93 0.999 1.00 Glioma 40 0.985 0.68 0.996 0.88 Liver 348 0.988 0.70 0.979 0.84 Biliary Tract 40 0.996 0.85 0.998 0.98 Bladder 392 0.996 0.95 0.998 0.98 Breast 135 0.946 0.10 0.378 0.00 Colorectal 155 0.99 0.83 0.955 0.86

Citation Format: Grace Zhou, Hai Hu. Development and validation of a serum microRNA-based diagnostic model for early detection of multiple cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3352.
Abstract 338: Regulation of gap junction intercellular communication in pancreatic cancer cells
Authors: Nguyen Annelise , Doan My
Published: April 2023

According to the American Cancer Society, approximately 62,210 people (32,970 men and 29,240 women) in the United States are estimated to be diagnosed with pancreatic cancer by the end of 2022. An estimated 80% of these patients will die as a result of pancreatic cancer. This type of cancer has been challenging to treat due to the complexity of tumor cells, tumor aggressiveness, current drug availability, and drug delivery. A new class of substituted quinolines, known as PQ compounds, acts as gap junction enhancer, having the ability to increase the gap junctional intercellular communication cancerous cells. The goal of this project is to provide a new approach to treat pancreatic cancer via restoration of gap junctional intercellular communication and combining treatment with antineoplastic drugs. One approach was to increase cell communication of pancreatic cancer cells using small molecules, PQs, and hence allowing antineoplastic drugs to effectively travel from cell to cell. Pancreatic cancer cells were treated with gap junction enhancers, PQs, in the presence and absence of antineoplastic drugs. Interestingly, 5-FU significantly decreased the expression of gap junction protein, connexin 43, compared to control and PQ1 treatments. PQ1 has demonstrated to increase Cx43 expression and increase 4.5-fold of gap junction activity. Furthermore, cells treated with nilotinib have a significant effect on PANC-1 cells. Current studies focus on functional assays to determine if PQ1 can increase cell-cell communication, allowing a reduced concentration of antineoplastic drugs to be used. Overall, the findings provide an initial insight to a new approach in treating pancreatic cancer.

Citation Format: Annelise Nguyen, My Doan. Regulation of gap junction intercellular communication in pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 338.
Abstract 3382: Detection of CTCs using a micro-channel combined fluid dynamics and electrophoresis in colorectal cancer
Authors: Ohta Ryo , Yamada Takeshi , Taniai Nobuhiko , Yoshida Hiroshi
Published: April 2023

Background: Circulating tumor cells (CTCs) are tumor cells that can be isolated from peripheral blood. Investigation of CTCs might allow comprehensive studies at the DNA, RNA, and protein. However, fewer CTCs generally can be collected from patients with colorectal cancer, limiting the possibility of clinical applications of CTC analysis despite the development of many different methods. We have developed a micro-channel device that integrates hydrodynamic filtration (HDF) capable of size separation and dielectrophoresis (DEP) capable of electrical property separation, using peripheral blood from colorectal cancer patients.

Method: A separation test was performed using a whole peripheral blood sample obtained from colorectal cancer patients with KRASmutation in the tumor tissue. The sample blood was diluted 2-fold with PBS, and red blood cells and plasma were removed using HDF. After that, Leukocytes and CTCs were separated in the same channel by DEP. DNA was extracted from CTCs (port 1) and leucocyte fraction (port 2), and KRAS mutations were detected using droplet digital PCR. In order to evaluate the separated fractions, microwell slide was used for evaluation by nuclear staining, CD45 and EpCAM antibody staining.

Results: 52 colorectal cancer patients were participated. The median age of patients was 71 years, 38 (73%) were male. Twenty-eight (53.9%) had colon cancers. Four stage III and 48 stage IV patients were included. The quality of DNA extracted from all patients were suitable for molecular genetic analysis. The DNA concentration of port 1 was significantly lower than that of port 2 (371 ng/mL vs 36667 ng/mL, P<0.001). Using droplet digital PCR, KRAS mutations were detected in the port 1 of 8 (15.4%) patients and in port 2 of 15 (28.8%) patients. In 35.5% of patients, KRAS mutations were detected in either port 1 or port 2.

Conclusion: This device does not require pretreatment and is easy to extract CTCs. Since CTCs are contaminated with the leukocyte fraction, it is necessary to improve the accuracy of CTCs collection by adjusting the conditions of DEP.

Citation Format: Ryo Ohta, Takeshi Yamada, Nobuhiko Taniai, Hiroshi Yoshida. Detection of CTCs using a micro-channel combined fluid dynamics and electrophoresis in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3382.
Abstract 341: Critical role of defensins in respiratory viral infection induced breast cancer progression
Authors: Ali Umer , Tiriveedhi Venkataswarup
Published: April 2023

Respiratory viral infections (RVI) such as influenza and COVID19 impact the host systemic immune system along with causing deleterious chronic inflammatory responses and respiratory distress. While the role of chronic inflammation in cancer is well-established, the role of RVI on tumorigenesis is poorly defined. To study the role of RVI on breast cancer, we first infected murine respiratory epithelial cells (mRES) with murine sendai virus (mSV), an analog for human parainfluenza virus. These infected mRES were co-cultured with 4T1 murine breast cancer cells in 1:1 dilution on a single 2D plate and also in trans-well format. Both in co-culture and transwell culture we saw a 40-80% (p<0.05) increased proliferation of breast cancer cells. Similarly, when 4T1 cells were treated with the supernatant collected from infected mRES cells in 1:5 dilution, also demonstrated a 2.3 fold increased breast cancer cell proliferation. The cytokine analysis from the supernatant collected from infected mRES cells demonstrated a 17-23 fold enhanced secretion of α/β-defensins. Direct treatment of α-defensin (cyptidin-4, 10 pg/mL) and β-defensin-3 (mBD3, 20 pg/mL) on 4T1 cells demonstrated enhanced expression of chemokine metastatic receptor, CXCR4 (4.3 fold), angiogenic factor, VEGF (12.8 fold) and cell division favoring factor, CDK2 (8.1 fold). Further, analysis of infected mRES cells demonstrated upregulation of toll-like receptor 2 (TLR2) and NOD-like receptor protein 3 (NLRP3) expression. Interesting, co-cultured of infected mRES with syngeneic murine CD4 T cells induced exhaustion phenotype (PD1+ and CTLA4+) differentiation of CD4 T cells. Taken together, these data suggest that respiratory viral infections through induction of cancer cell proliferation and inhibiting anti-tumor adaptive immune responses promote breast cancer proliferation.

Citation Format: Umer Ali, Venkataswarup Tiriveedhi. Critical role of defensins in respiratory viral infection induced breast cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 341.
Abstract 3410: AVATASCAN®, a pioneer of functional precision medicine in guiding clinical decision-making
Authors: Her Nam-Gu , Lee Gi Ju , Hyun Seung Yoon , Park San Ha , Ahn Jae Woo , Kang Ji Soo , Toh Hong Boon , Nam Do-Hyun
Published: April 2023

Precision medicine refers to the tailoring of individual therapeutics based on each patient’s genetic, phenotypic, and clinical characteristics, therefore seeking the most effective treatment for the patient. With recent advances in genome sequencing technology, it was anticipated that identifying specific genetic alterations would contribute greatly to the realization of precision medicine. However, most cancer patients do not benefit from genomic precision medicine, as shown in recent Next Generation Sequencing (NGS)-driven clinical trials. Functional precision medicine directly uses patient tumor cells to test their ex vivo responses to diverse drugs to predict the most effective drugs. Functional precision medicine is emerging because it provides immediate translatable information to select drugs among clinically available therapeutics. AVATASCAN®, developed by Samsung Medical Center and AimedBio Inc., is a robust and accurate high throughput functional precision medicine platform with more than 1,500 historical sample data. AVATASCAN® has tested more than 1,500 cancer patient samples across 14 different tumor types, including glioblastoma, lung, colorectal, stomach and breast cancers. We achieved a tissue culture success rate and a drug screening success rate of more than 90% in most tumor types. A retrospective analysis of the clinical outcomes of patients given matched drugs demonstrated actual complete/partial response in 85% of AVATASCAN® screening responders. Recognizing its potential for clinical application, AVATASCAN® was selected as one of the datasets referenced by the pediatric tumor board at Seoul National University Hospital for clinical decision-making. Furthermore, AVATASCAN® is now available in Singapore and Thailand as an early-access, premium precision medicine service. Overall, we present compelling evidence that AVATASCAN® is a valuable platform for personalized cancer therapy. Therefore, we are moving forward to adopt and expand this platform in clinical applications.

Citation Format: Nam-Gu Her, Gi Ju Lee, Seung Yoon Hyun, San Ha Park, Jae Woo Ahn, Ji Soo Kang, Hong Boon Toh, Do-Hyun Nam. AVATASCAN®, a pioneer of functional precision medicine in guiding clinical decision-making [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3410.
Abstract 3411: Proof of concept study of next generation drug screening platform for glioma patients
Authors: Her Nam-Gu , El-Heliebi Amin , Lee Gi Ju , Park San Ha , Prietl Barbara , Ahn Stephanie S. , Toh Hong Boon , Pieber Thomas R. , Nam Do-Hyun
Published: April 2023

Precision medicine endeavors to match patients to therapies mostly on the basis of the genomic alterations in their tumors. However, it has been shown that ‘genomics only’ approach is often insufficient for best drug selection. Functional precision medicine has been recently emerging as it provides directly translatable information on which drug to choose among available drugs. Traditionally, cancer drugs are tested in cancer cell line models, but cell lines cannot represent individual clinical patients and are too biologically pronounced to be useful for drug screening purposes. Our patient-derived cell (PDC) drug screening system overcomes these points and can generally also correlate with genome changes. AVATAMED and CBmed, together with AimedBio, are collaborating in a Precision Cancer Project, set out next generation drug screening platform to screen Austrian glioma patients for proof-of-concept study. Fresh native tissue from both low- and high-grade glioma are collected. Tumor tissue is used for PDC isolation using mechanical and enzymatic tissue dissociation. To preserve the original tumor similarity, tissue is short term cultured under two weeks, and PDCs are seeded and treated with a panel of clinical- and preclinical drugs followed by viability assessment. We have cultured 65 PDCs from both low- and high-grade glioma. Tissue culture success rate and drug screening success rate was >85% suggesting that specimen logistics and tissue processing are well-established and conducted. PDCs were treated with 79 drugs that are commonly used in clinical setting. In order to evaluate the quality of the results, the screening was conducted at two different sites, AimedBio and CBmed, and the data were compared. Overall, high accuracy and reproducibility were confirmed by low Coefficient of Variation (CV) between replicates and high Z-factor between negative and positive controls. In addition, data generated from both sites showed high correlations. Collectively, our next-generation drug screening platform shows the potential to give the best therapeutic options to glioma patients. To this end, we are moving forward use this platform in clinical study.

Citation Format: Nam-Gu Her, Amin El-Heliebi, Gi Ju Lee, San Ha Park, Barbara Prietl, Stephanie S. Ahn, Hong Boon Toh, Thomas R. Pieber, Do-Hyun Nam. Proof of concept study of next generation drug screening platform for glioma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3411.
Abstract 3426: Catalytic degraders effectively address kinase site mutations in EML-ALK oncogenic fusions
Authors: Gao Yang , Jiang Baishan , Kim Hellen , Che Jianwei , Donovan Katherine , Hatcher John , Huerta Fidel , Kwiatkowski Nicholas , Liu Yingpeng , Liuni Peter , Metivier Rebecca J. , Murali Vineeth , Nowak Radosław , Zhang Tinghu , Fischer Eric , Gray Nathanael , Jones Lyn
Published: April 2023

Heterobifunctional degraders, known as proteolysis targeting chimeras (PROTACs), is an emerging modality for drug discovery, and theoretically possess catalytic mode-of-action, yet few studies have either confirmed or exploited this potential advantage of event-driven pharmacology. Degraders of oncogenic EML4-ALK fusions were developed by conjugating ALK inhibitors to cereblon ligands guided by computational models. To study the sub-stoichiometric capacity of the degrader molecules, simultaneous optimization of pharmacology and compound properties using ternary complex modeling and physicochemical considerations yielded multiple catalytic degraders that were more resilient to clinically relevant ATP-binding site mutations than parental kinase inhibitor drugs. Using HiBiT assay to assess the target degradation and NanoBRET for target occupancy, we illustrated the concept of catalytic degradation of EML4-ALK. Our strategy culminated in the design of the orally bioavailable derivative CPD-1224 that avoided hemolysis (a feature of detergent-like PROTACs), degraded the otherwise recalcitrant compounded mutant L1196M/G1202R in vivo, and commensurately slowed tumor growth, while the third generation ALK inhibitor drug lorlatinib had no effect. These results validate our original therapeutic hypothesis by exemplifying opportunities for catalytic degraders to proactively address binding site resistant mutations in cancer.

Citation Format: Yang Gao, Baishan Jiang, Hellen Kim, Jianwei Che, Katherine Donovan, John Hatcher, Fidel Huerta, Nicholas Kwiatkowski, Yingpeng Liu, Peter Liuni, Rebecca J. Metivier, Vineeth Murali, Radosław Nowak, Tinghu Zhang, Eric Fischer, Nathanael Gray, Lyn Jones. Catalytic degraders effectively address kinase site mutations in EML-ALK oncogenic fusions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3426.
Abstract 3452: TNG908, a brain-penetrant MTA-cooperative PRMT5 inhibitor, is efficacious in preclinical glioblastoma models
Authors: Zhang Minjie , Tsai Alice , Cottrell Kevin M. , Haines Brian B. , Wilker Erik , DiBenedetto Heather , Weitzman Ron , Huang Alan , Davis Charles B. , Maxwell John P. , Briggs Kimberly J.
Published: April 2023

Glioblastoma multiforme (GBM) is one of the most aggressive primary malignant brain tumors in adults. The median overall survival (OS) of GBM patients is poor (1-2 years) on standard of care therapies, which include surgery, radiotherapy, and the alkylating agent temozolomide. Therefore, developing new treatments for GBM is an urgent unmet medical need. TNG908 is a clinical stage MTA-cooperative PRMT5 inhibitor that inhibits PRMT5 activity selectively in MTAP-null cells leveraging a synthetic lethal interaction. Approximately 40% of GBM tumors are homozygous null for MTAP [1, 2]. In preclinical in vitro studies, the antiproliferation activity of TNG908 was 15 times more potent in MTAP-null cancer cell lines than in MTAP WT cells. TNG908 has high passive permeability and is not a substrate for P-glycoprotein nor Breast Cancer Resistant Protein efflux transporters, attributes which are favorable for crossing the blood-brain barrier. TNG908 demonstrated in vivo brain penetration in multiple preclinical studies, including non-human primates and mice, suggesting that TNG908 may fulfill the significant unmet need in the treatment of GBM. Pharmacodynamic activity of TNG908 to inhibit PRMT5 was demonstrated by decreased SDMA-modified protein levels in a dose-dependent manner in a GBM xenograft model. TNG908 demonstrated dose-dependent antitumor activity in multiple hyper- and hypomethylated GBM subcutaneous models, including cell lines and patient-derived xenograft models. Despite reduced Kpuu in rodents compared to primates, oral administration of TNG908 drove near tumor stasis and increased median survival by 3-fold in a highly aggressive murine GBM orthotopic model. In summary, TNG908 is a potent, brain-penetrant small molecule PRMT5 inhibitor that is selective for MTAP-null tumor cells with good drug-like properties and strong antitumor activity in preclinical models of GBM. As such, TNG908 may provide a novel treatment strategy for MTAP-deleted GBM patients. *MZ and AT contributed equally to the work.

Reference: 1.Cerami et al., Cell, 2013. 2.Gao et al., Sci Signal, 2013

Citation Format: Minjie Zhang, Alice Tsai, Kevin M. Cottrell, Brian B. Haines, Erik Wilker, Heather DiBenedetto, Ron Weitzman, Alan Huang, Charles B. Davis, John P. Maxwell, Kimberly J. Briggs. TNG908, a brain-penetrant MTA-cooperative PRMT5 inhibitor, is efficacious in preclinical glioblastoma models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3452.
Abstract 3551: Single-cell characterization of pediatric T-cell acute lymphoblastic leukemia transcriptomes
Authors: Pushel Irina , Yoo Byunggil , Louiselle Daniel , Gibson Margaret , Farooqi Midhat S. , August Keith
Published: April 2023

Pediatric T-cell acute lymphoblastic leukemia (T-ALL) is a rare cancer with a diverse set of causative genomic abnormalities. The variety and limited characterization of genomic mechanisms underlying this disease presents a unique challenge for the development of improved treatments for these patients. Using the 10x Genomics Chromium system, we have generated single-cell gene expression (scRNA) and chromatin accessibility (scATAC) data for blood and/or bone marrow samples collected at diagnosis from eight pediatric T-ALL patients. We sequenced 21,669 high-confidence cells and performed normalization and downstream analysis using the Seurat v4.0.2 package in R 4.0.3. Our data show that patient cancer cells show highly individual transcriptomes and are often characterized by combinatorial expression patterns of genes including MEF2C, BCL11B, HOXA9, NKX2-1, and LMO2, which have been previously implicated for their roles in various T-ALL subtypes. Additionally, we observe that each patient has a unique ratio of cancer cells with a CD4+ T-cell, CD8+ T-cell, and hematopoietic stem cell-like identity, which may be linked to treatment response and patient outcome. This dataset represents one of the first forays into exploring the biology of pediatric T-ALL at the single-cell level. Our comparative analysis of patient cancer cells reveals novel insights into the transcriptional and chromatin-related for distinct subsets of pediatric T-ALL cases, paving the way for future treatment selection and drug development to improve outcomes for these patients.

Citation Format: Irina Pushel, Byunggil Yoo, Daniel Louiselle, Margaret Gibson, Midhat S. Farooqi, Keith August. Single-cell characterization of pediatric T-cell acute lymphoblastic leukemia transcriptomes. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3551.
Abstract 3569: An animal model of GLI2-amplified medulloblastoma
Authors: Pei Yanxin
Published: April 2023

Medulloblastoma (MB) is the most common malignant brain tumor affecting children. Among the known subtypes of MB, GLI2-amplified SHH-MB associated with P53 mutations has the worst prognosis, and a 5-year survival rate of less than 30%. Moreover, GLI2-amplified MBs are non-responsive to SMO inhibitors, the only targeted treatment option available for SHH-MB. To address this treatment gap, there is an urgent need to identify novel potential therapeutic targets, an effort that requires better understanding of key cellular and molecular mechanisms underlying GLI2-amplified MB. To that end, we generated a mouse model of GLI2-amplified MB and identified GL12 as the critical driver of tumorigenesis, and granule cell progenitors (GCPs) as the cells of tumor origin. We further found that GLI2 specifically drives embryonic, but not neonatal, Math1+ GCPs to form SHH-MB tumors. An scRNA-seq analysis revealed MAPK pathway activation specifically in embryonic Math1+ GCPs. The MAPK pathway was activated in mouse and human GLI2-driven MB tumors, and a MEK/ERK inhibitor significantly inhibited GLI2-driven MB in vivo. Based on these findings, we hypothesize that GLI2-driven MB originates from a specific population of Math1+ GCPs during a discrete temporal interval in cerebellar development, and that targeting MAPK/MEK/ERK pathway components represents a novel approach to treatment of GLI2-amplified MB.

Citation Format: Yanxin Pei. An animal model of GLI2-amplified medulloblastoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3569.
Abstract 3580: Noninvasive immuno-PET imaging of CD8+T cell behavior in tumor bearing mice models treated with SAR444245
Authors: d'Heilly Sebastion , Tirode Fabrice , Gaulin Maxime , Anne Dudzicki , Bluet Guillaume , Guerif Stéphane , Descamps Francis , Casteels Peter , Li Xiangming , Wang Rui , Meng Robin , Jouannot Erwan
Published: April 2023

SAR444245 (formerly THOR-707) is a site-specific pegylated recombinant human IL-2 molecule which blocks IL-2R alpha-binding while retains near-native affinity for beta/gamma IL-2 receptor subunits. SAR444245 reduces B16-F10 tumor proliferation in C57BL/6 mice and induces a short transient migration of CD8+ T and NK cells from peripheral blood to lymphoid organs before a strong proliferation (Ptacin et al, Nat Commun 2021). Here we report CD8+ T cells imaging post SAR444245 as monotherapy in naive and tumor bearing mice using nuclear imaging. An anti-murine CD8+ PET probe was injected in mice 24 hours post SAR444245 administration and nuclear imaging was performed at different times over 6 days. In C57BL/6 naive and in poorly immunogenic B16-F10 tumor bearing animals, images reveal a decreased Standardized Uptake Value (SUV) within blood in treated animals at 3mg/kg compared to control ones. In parallel, blood SUV Ratio (SUVR) increases within spleen, lymph nodes and thymus after SAR444245 injection compared to control mice. On tumor site, a stronger SUVR is measured at 144 hours post injection in SAR444245 treated group than in the control group. A dose escalation ranging from 1 to 6 mg/kg of SAR444245 was performed in highly immunogenic CT26 tumor bearing animal. Blood SUV decreases in the treated groups compared to the control group. In lymphoid organs, all SUVR increase at 144 hours post injection in the treated groups compared to the control one. In CT26 tumors, a slight increase of the SUVR is observed for groups treated at 3 and 6 mg/kg. In parallel, a pronounced reduction of tumor growth is observed in the treated group at 6mg/kg. These studies non-invasively confirm the mode of action of SAR444245 with a rapid relocation of CD8+ T cells from peripheral blood to lymphoid organs and tumor site.

Citation Format: Sebastion d'Heilly, Fabrice Tirode, Maxime Gaulin, Dudzicki Anne, Guillaume Bluet, Stéphane Guerif, Francis Descamps, Peter Casteels, Xiangming Li, Rui Wang, Robin Meng, Erwan Jouannot. Noninvasive immuno-PET imaging of CD8+T cell behavior in tumor bearing mice models treated with SAR444245. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3580.