bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3947: S100A8/A9-RAGE pathway and chronic airway inflammation in cigarette smoke-induced lung carcinogenesis
Authors: Kang Hye Seon , Kim In Kyoung , Yeo Chang Dong , Kim Jin Woo , Lee Sang Haak
Published: April 2023

Background: Cigarette smoke-induced chronic airway inflammation increases the risk of lung cancer, and plays a multifaceted role in lung cancer initiation and progression, and RAGE-ligand axis (HMGB1, S100A8/A9) contributes Cigarette smoke-induced persistent and progressive inflammation. The aim of the present study was to determine the inflammatory effect on S100A8/A9-RAGE pathway in smoke-induced carcinogenesis in lung cancer.

Methods: Human alveolar adenocarcinoma cells (A549) and normal bronchial epithelial cells (BEAS-2B) were co-cultured for 24 h with the PBMC and then, cells were incubated in the presence or absence of 1.5% cigarette smoke extract (CSE) for 24 h. The cells transfected with siRNA targeting S100A8/9 or RAGE. The cell viability, colony-forming ability, migration, invasion, metastasis and morphological changes were assessed. Female A/J mice were given a benzo(a)pyrene (100mg/kg) in 0.1 ml corn oil via oral gavage at once a week for 3 weeks, and administrated CSE (1.25 ul/g) via intratracheal (IT) at twice a week for 4 weeks. Tumor load was determined by averaging the total tumor volume in each group. BAL differential cells were counted to analyze inflammatory cells.

Results: Cellular growth and colony-forming were promoted by exposure to CSE, and significantly inhibited by S100A8/9 and RAGE-siRNA transfection, especially in A549 cells than BEAS-2B. Cell migration, invasion, MMP-2/9 activities, and the mRNA and protein expression of TLR4, NF-κB, RAGE, S100A8/9 and HMBG1 were higher in CSE exposed A549 cells. S100A8/9 and RAGE-siRNA transfection attenuated those expression, significantly. B(a)P induced an average tumor volume 7.28 mm3 per mouse, and CSE significantly increased tumor volume (8.10 mm3) compared to the B(a)P group. The total cell number and lymphocytes in BAL fluids tended to increase after CSE administration in lung cancer mouse group. Moreover, CSE significantly induced the levels of S100A8/9 and RAGE, p38 and p-ERK, and decreased JNK in serum and tumors with adjacent lung tissues (p < 0.05).

Conclusions: We identified cigarette smoke-induced S100A8/A9-RAGE pathway promoted airway inflammation and lung carcinogenesis in vitro and in vivo. Although S100A8/A9 protein contributes to many types of disease, this study will help us gain a better understanding of the complexities of cancer progression. Further studies are needed for therapeutic target of cigarette smoke-induced inflammation and immunosuppression in tumor microenviornment.

Citation Format: Hye Seon Kang, In Kyoung Kim, Chang Dong Yeo, Jin Woo Kim, Sang Haak Lee. S100A8/A9-RAGE pathway and chronic airway inflammation in cigarette smoke-induced lung carcinogenesis. [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 3947.
Abstract 3963: Mass spectrometry reveals a downregulation of calreticulin after neoadjuvant chemotherapy in pancreatic cancer
Authors: Stoffel Michael C. , Dingendorf Elisabeth BH , Kittel Dahlia L. , Kristiansen Glen , Lerbs Tristan
Published: April 2023

Introduction: Pancreatic ductal adenocarcinomas are one of the deadliest cancer types with limited response to current therapies. Thus, this study examined changes in the proteome of epithelial cells in pancreatic cancer after neoadjuvant chemotherapy with gemcitabine, to identify new therapeutic targets.

Methods: Upon receiving patient consent, fresh pancreatic cancer tissue was received from surgical tumor resections at the University Hospital of Bonn. We mechanically and enzymatically digested and purified the tissue to create single-cell suspensions. After staining the cells against CD45 und CD326, we purified epithelial cells, blood cells, and stroma through FACS. To analyze their proteome, the sorted cells were lysed in TEAB buffer and the protein lysates were measured by the mass spectrometry core facility at the University of Bonn. We then analyzed the data using DAVID and Shiny GO.

Results: Gene enrichment analyses showed a significantly reduced expression of calreticulin after neoadjuvant chemotherapy in epithelial cells. A direct comparison between untreated and treated epithelium cancer cells using a heat map showed a decreased expression of actin and enzymes of glycolysis in treated cancer cells. Moreover, untreated epithelial cells showed an increase of proteins related to MHC class Ib protein complex assembly and antigen processing and presentation of endogenous peptide antigen via MHC class. In contrast, treated cells showed a higher presence of proteins for nutrient supply from the blood. Furthermore, untreated stroma cells exhibited an increased number of proteins responsible for mesenchymal migration and for actin filament fragmentation.

Conclusion: This study shows that neoadjuvant chemotherapy with gemcitabine leads to distinct changes in the proteome of tumor cells in pancreatic cancers. In particular, a decreased expression of the “eat-me-signal” calreticulin impairs macrophage-mediated cancer cell phagocytosis, thereby indicating a mechanism by which cancer cells protect themselves after chemotherapy.

Citation Format: Michael C. Stoffel, Elisabeth BH Dingendorf, Dahlia L. Kittel, Glen Kristiansen, Tristan Lerbs. Mass spectrometry reveals a downregulation of calreticulin after neoadjuvant chemotherapy in pancreatic 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 3963.
Abstract 3964: APE1 protects SMAD3 against ROC1 ubiquitin mediated degradation in esophageal adenocarcinoma cells
Authors: Ballout Farah , Lu Heng , Peng Dunfa , El-Rifai Wael
Published: April 2023

Background: The incidence of esophageal adenocarcinoma (EAC) has increased more than six-fold over the past three decades and continues to rise in the Western world. The 5-year survival rate for EAC patients is less than 20% which underscores the need to better understand the underlying mechanisms to identify new therapeutic approaches. This study aimed at investigating the potential role of APE1 in regulating SMAD3 and promoting EAC progression.

Methods and Results: Western blot data showed that APE1 and SMAD3 were highly expressed in EAC cell lines. APE1 silencing reduced SMAD3 nuclear expression and downregulated its downstream targets SERPINE1 and c-myc. These results were confirmed by immunofluorescence staining showing loss of nuclear accumulation of SMAD3 after APE1 knockdown. Mechanistically, immunoprecipitation and proximity ligation assays revealed a direct binding between APE1 and SMAD3 in the nucleus. Further investigation showed that APE1 binds to the C-terminal MH2 domain of SMAD3, and this binding protects SMAD3 from ubiquitin mediated proteasomal degradation by blocking its interaction with the RING finger protein, ROC1. Interestingly, APE1-redox-specific inhibition (APX2009) downregulated SMAD3 expression and the APE1 redox-deficient mutant (C65A) disrupted APE1-SMAD3 binding indicating that this regulation depends on APE1 redox activity.

Conclusion: Our findings establish a role of APE1 in regulating SMAD3 in EAC. These findings provide a potential therapeutic approach for the treatment of EAC by the pharmacological inhibition of APE1.

Citation Format: Farah Ballout, Heng Lu, Dunfa Peng, Wael El-Rifai. APE1 protects SMAD3 against ROC1 ubiquitin mediated degradation in esophageal adenocarcinoma 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 3964.
Abstract 3978: The growth inhibitory effect of MDV3100 on hormone receptor-positive, HER2-negative breast cancer cell lines with low DYRK2 expression
Authors: Imawari Yoshimi , Mimoto Rei , Yoshida Kiyotsugu
Published: April 2023

Introduction: Tumor progression is the main cause of death in patients with breast cancer. Accumulating evidence suggests that dual-specificity tyrosine-regulated kinase 2 (DYRK2) functions as a tumor suppressor by regulating cell survival, differentiation, proliferation and apoptosis. Recently, we reported for the first time that DYRK2 inhibits breast cancer stem cells and tumor cell proliferation through transcriptional downregulation of KLF4 and CDK14, respectively.Our previous study showed that Androgen receptor (AR) was a DYRK2-dependent transcriptional activator. In this context, we hypothesized that MDV3100, an inhibitor of AR, may be effective for breast cancer with low DYRK2 expression.

Methods: We established stable DYRK2-depleted cells. MCF-7 cells, hormone receptor-positive, HER2-negative breast cancer cell line, were transfected with pSuper-puro vector (pSuper control) or pSuper-puro DYRK2 shRNAs (shDYRK2) along with puromycin to isolate stable cell lines.These manipulated cells were compared with the control cells by various assays. We analyzed the activity of AR by Androgen receptor reporter assay. MTS assays were used to assess growth inhibition after treatment with MDV3100.

Results: Androgen receptor reporter assay showed that knocking down DYRK2 induced AR transcription activity in MCF-7 cells.In MCF-7 DYRK2-depleted cells, MTS assays revealed that the treatment by MDV3100 was more effective than that in MCF-7 control cells.

Conclusion: These findings demonstrate that reduced DYRK2 expression in hormone receptor-positive, HER2-negative breast cancer increases the activity of AR and significantly inhibits the growth by treatment with MDV3100.

Citation Format: Yoshimi Imawari, Rei Mimoto, Kiyotsugu Yoshida. The growth inhibitory effect of MDV3100 on hormone receptor-positive, HER2-negative breast cancer cell lines with low DYRK2 expression. [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 3978.
Abstract 3983: Defining the role of FANCA in breast cancer development and cell cycle progression
Authors: Luo Liang , Liu Wenjun , Palovcak Anna , Yuan Fenghua , Li Fang , Calkins Daniel , Li Yan , Briegel Karoline , Bilbao Daniel , Roberts Evan , Mason Christian , Liu Zhao-Jun , Daunert Sylvia , Zhang Yanbin
Published: April 2023

FANCA is one of the 22 known Fanconi anemia (FA) pathway genes and is indispensable for interstrand crosslink (ICL) repair. It is reported that FANCA is responsible for about 64% of FA cases and one important clinical characterization of FA patients is predisposition to cancer. However, analysis of TCGA database reveals that FANCA level is elevated in breast cancer patients, especially in ER- breast cancer, which is associated with the methylation status at the S-shore of CpG island ahead of FANCA gene. Moreover, the FANCA level is negatively correlated with the survival rate of breast cancer patients. To understand the implication of FANCA in breast cancer development, we have targeted FANCA by either CRISPR mediated knock-out (KO) or shRNA mediated knockdown (KD) in MDA-MB-231 breast cancer cells. Our data suggest that depletion of FANCA protein inhibits breast cancer growth both in vitro and in vivo. On the other hand, overexpressing FANCA in low-FANCA breast cancer cell line MDA-MB-468 promotes cancer cell proliferation. Cell cycle profiling reveals an inefficiency of MDA-MB-231 FANCA KO cells in G1 to S transition due to p21 and p27 upregulation in FANCA KO cells. In addition, the proliferation inefficiency in MDA-MB-231 FANCA KO cells is caused by Rb hypophosphorylation. Therefore, we conclude that FANCA contributes to the breast cancer development by promoting cell cycle progression through Rb/E2F signaling pathway. Our immunoprecipitation (IP) results indicate that in MDA-MB-231, FANCA is interacting with HES1, which is a transcription suppressor that binds to the promoter region of CDKN1A and CDKN1B to promote cell cycle progression, explaining how FANCA participates in cell cycle progression in breast cancer.

Citation Format: Liang Luo, Wenjun Liu, Anna Palovcak, Fenghua Yuan, Fang Li, Daniel Calkins, Yan Li, Karoline Briegel, Daniel Bilbao, Evan Roberts, Christian Mason, Zhao-Jun Liu, Sylvia Daunert, Yanbin Zhang. Defining the role of FANCA in breast cancer development and cell cycle 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 3983.
Abstract 3988: The role of HER3 mutations in the progression of colon cancer and modulation of drug sensitivity and resistance
Authors: Kilroy Mary K. , Mishra Rosalin , Stupecki Anastasia , Feroz Wasim , Alanazi Samar , Garrett Joan T.
Published: April 2023

Colorectal cancer is a disease of the colon and rectum that will claim about 52,580 American lives in2022. The most common treatment of colorectal cancer is surgery plus chemotherapy, although thereare some FDA approved targeted therapies such as regorafenib (targeting VEGF) or cetuximab (targetingEGFR). EGFR, along with HER2, HER3, and HER4 are members of the HER family of receptor tyrosinekinases that upon homo- or heterodimerization activate downstream signaling, growth, and survivalpathways. In recent years, more attention has been paid to HER2 and HER3’s roles in colorectal cancers,as they may cause resistance to targeted therapies. OWe are investigating naturally occurring mutantHER3’s role in colorectal cancer, as about 6% of all colorectal cancers contain a HER3 mutation. We arecurrently investigating how HER3 mutations may affect sensitivity to current therapies through HERfamily receptor dimerization and be involved in tumor metastasis. We are assessing the degree to whichmutant and wild-type HER3 have the ability to dimerize with EGFR, HER2, MET, and IGF1R the resultingeffects in colon cancer, as these receptor tyrosine kinases are HER3 binding partners. As HER3 bindingpartners are diverse, it may be that the pathway HER3 and its binding partner activate may influencetreatment strategy. It has been noted that mutant HER2 and HER3 could confer sensitivity to HER familyinhibitors, i.e. afatinib, in bladder cancer, and we have seen a difference in IC50 values of afatinibbetween cell lines containing wild-type or mutant HER3. If mutant HER3 is involved in therapeuticresistance or sensitivity and tumor progression, our findings may present a new biomarker for targetedtreatments in colorectal cancer with the eventual goal of increased overall patient survival.

Citation Format: Mary K. Kilroy, Rosalin Mishra, Anastasia Stupecki, Wasim Feroz, Samar Alanazi, Joan T. Garrett. The role of HER3 mutations in the progression of colon cancer and modulation of drug sensitivity and resistance. [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 3988.
Abstract 3990: Imaging molecular alterations during tucatinib response in preclinical models of HER2+ breast cancer
Authors: Song Patrick , Mansur Ameer , Sorace Anna
Published: April 2023

Introduction: Human epidermal growth factor receptor 2 (HER2) is overexpressed in 25% of breast cancers. Tucatinib, a small molecule HER2 inhibitor, was FDA approved for inoperable or metastatic HER2+ breast cancer. As many of these patients have inoperable tumors, there is a need to identify imaging metrics that can characterize response to tucatinib. Positron emission tomography (PET) imaging can quantify changes in the tumor microenvironment that precede changes in tumor size through imaging with radiopharmaceuticals that target proliferation (fluorothymidine, [18F]-FLT), hypoxia (fluoromisonidazole, [18F]-FMISO) and HER2 expression ([89Zr]-Pertuzumab). The goal of this study is to use advanced PET imaging to non-invasively monitor response to tucatinib in HER2+ primary breast cancer and quantify the subsequent tumor microenvironment modulation.

Methods: HER2+ cell line (BT474) and patient derived xenograft (BCM 3472) tumor models were engrafted and developed to 270 ± 166.4 mm3 before being enrolled into experiments. Mice were treated with 50 mg/kg tucatinib via PO and were imaged with [18F]-FLT PET (N = 8) on days 0, 3 and 7, [18F]-FMISO PET (N = 6) on days 0, 3 and 7, or [89Zr]-Pertuzumab PET (N = 5) on days 0 and 14. Intratumoral proliferation, hypoxia and HER2 expression were quantified with standardized uptake value (SUV). Following the final imaging timepoint, tumors were excised for immunohistochemistry against Ki-67 (proliferation), pimonidazole (hypoxia), and HER2. A non-parametric T-test was used to assess for significance.

Results: Tucatinib treated BT474 and BCM3472 tumors had a 2.07 and 2.63 fold decrease in tumor volume, respectively (p<0.01). Tucatinib treated BT474 and BCM3472 tumors had significantly decreased hypoxia and proliferation, relative to control tumors (p<0.05). Tucatinib treated BT474 tumors had significantly decreased HER2 expression (p<0.05); however, no significant change in HER2 expression was observed in tucatinib treated BCM3472 tumors.

Conclusion: Tucatinib significantly decreases tumor volume and decreases intratumoral proliferation and hypoxia in both cell-line and patient-derived xenograft models of HER2+ breast cancer. Our data suggests molecular imaging may drive understanding of and predict response to tucatinib therapy.

Acknowledgements: Tucatinib was provided by Seagen Inc. Bothell, Washington, USA.

Citation Format: Patrick Song, Ameer Mansur, Anna Sorace. Imaging molecular alterations during tucatinib response in preclinical models of HER2+ breast 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 3990.
Abstract 3993: In vivo efficacy of an anti-human IL11RA antibody in B-hIL11RA mice with APAP-induced liver damage
Authors: He Fang , Zhang Rufeng , Kuraguchi Mari , Zhao Lei , Yu Zhaoxue
Published: April 2023

Acetaminophen (N-acetyl-p-aminophenol; APAP) overdose is a common cause of drug-induced acute liver injury, thereby making APAP administration a frequently used experimental model due to its highly reproducible and dose-dependent hepatoxicity. APAP-induced liver injury (AILI) mouse models exhibited upregulated IL11 expression, indicating that targeting IL11 or its cognate receptor, IL11RA, could have therapeutic potential. To evaluate the in vivo efficacy of anti-human IL11RA antibodies in AILI mouse models, we first generated a human IL11RA knock-in mouse model (B-hIL11RA), in which human IL11RA replaced murine Il11ra, in situ. Human IL11RA mRNA and protein levels were detected in B-hIL11RA mice. We next examined whether therapeutic inhibition of IL11 signaling was effective in reducing AILI following administration of an anti-human IL11RA antibody. In vivo efficacy data showed that anti-human IL11RA antibody inhibited all aspects of AILI, including hepatocyte death (ALT, AST and TUNEL) and centrilobular necrosis. These data suggest that B-hIL11RA mice are an effective tool for preclinical in vivo efficacy evaluation of therapeutic anti-human IL11RA antibodies.

Citation Format: Fang He, Rufeng Zhang, Mari Kuraguchi, Lei Zhao, Zhaoxue Yu. In vivo efficacy of an anti-human IL11RA antibody in B-hIL11RA mice with APAP-induced liver damage. [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 3993.
Abstract 4032: Discovery of a potent, selective, and orally available SHP2 inhibitor that can penetrate blood-brain barrier
Authors: Kim Miyeon , Kim Dongsu , Yoon Kyeong Jin , Jeon Yeejin , Park Dohyun , Lee Mijung , Jeon Dahye , Jang Soyeon , Kim Jinhwan , Kim Eunji , Park Jihoon , Hong Victor , Lim Sang Kyun , Choi Sungpil
Published: April 2023

Src homology region 2-containing protein tyrosine phosphatase 2 (SHP2/PTPN11) is one of the key signaling proteins in many oncogenic receptor tyrosine kinase (RTK) pathways. It mediates RAS-driven downstream signaling and activates gene expression for tumorigenesis. Importantly, a critical role of SHP2 has been found during the process of temporal adaptation and ERK re-activation after the treatment of RAS/RAF/MEK/ERK inhibitors, which means that SHP2 could be a promising therapeutic target to shut down the oncogenic RAS pathway, especially in combination with RAS/RAF/MEK/ERK inhibitors. Here we present preclinical data of an allosteric small-molecule SHP2 inhibitor that is highly potent, selective, and brain-permeable. In vitro biochemical and cellular assays showed excellent potency against SHP2 protein. In addition, we confirmed exceptional in vivo activity in mouse xenograft models, and demonstrated combination effects with a KRAS G12C inhibitor. Most importantly, the BBB penetration and anti-tumor activity in brain was confirmed both in vitro and in vivo, supporting the great potential in treating brain-metastasized tumors. In summary, we have successfully discovered and developed a potent and blood-brain barrier permeable SHP2 inhibitor that provides a novel therapeutic option for diverse cancers associated in the RAS pathway.

Citation Format: Miyeon Kim, Dongsu Kim, Kyeong Jin Yoon, Yeejin Jeon, Dohyun Park, Mijung Lee, Dahye Jeon, Soyeon Jang, Jinhwan Kim, Eunji Kim, Jihoon Park, Victor Hong, Sang Kyun Lim, Sungpil Choi. Discovery of a potent, selective, and orally available SHP2 inhibitor that can penetrate blood-brain barrier. [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 4032.
Abstract 4036: NMS-0963 is a novel potent, selective and orally available Syk inhibitor with promising preclinical activity in diffuse large B-cell lymphoma
Authors: Saturno Grazia , Modugno Michele , Orsini Paolo , Cervi Giovanni , Buffa Laura , Motto Ilaria , Avanzi Nilla , Montemartini Marisa , Gasparri Fabio , Texido Gemma , Galvani Arturo , Isacchi Antonella
Published: April 2023

The B-cell receptor (BCR) is a key survival molecule for normal B cells and for most B-cell malignancies, such as Chronic Lymphocytic Leukaemia (CLL) and Non-Hodgkin’s Lymphomas (NHL) including Diffuse Large B-cell Lymphomas (DLBCL). Small molecule inhibitors of key signaling kinases involved in the BCR pathway, such as the Btk inhibitor ibrutinib and the PI3Kdelta inhibitor idelalisib, have already demonstrated significant clinical activity. Spleen tyrosine kinase (Syk) is a non-receptor cytoplasmic tyrosine kinase that plays a fundamental role in BCR signaling initiation thus representing an additional potential therapeutic target for the inhibition of the BCR pathway. Here, we report the discovery and characterization of NMS-0963, a novel potent, selective and orally available Syk inhibitor, identified through an integrated medicinal chemistry and rational design approach. NMS-0963 showed potent inhibitory effect on Syk in an in vitro biochemical assay and displayed a good selectivity profile on a broad panel of kinases. On-target potency was confirmed in a cell-based assay using BaF3 cells engineered to express constitutively activated Syk, showing potent inhibition of Syk activity resulting in antiproliferative effect with IC50s in the low nanomolar range. Effective inhibition of BCR-mediated signaling pathway was observed in DLBCL-derived cell lines. NMS-0963 showed favourable ADME profile and good in vivo pharmacokinetic profiles after oral administration in mouse, rat and dog. Furthermore, NMS-963 demonstrated striking in vivo efficacy in tumor models of CD79/Myd88 mutated DLBCL, superior to competitors. Together with a permissive preclinical safety profile, these results support a rational for clinical development of NMS-0963 in DLBCL.

Citation Format: Grazia Saturno, Michele Modugno, Paolo Orsini, Giovanni Cervi, Laura Buffa, Ilaria Motto, Nilla Avanzi, Marisa Montemartini, Fabio Gasparri, Gemma Texido, Arturo Galvani, Antonella Isacchi. NMS-0963 is a novel potent, selective and orally available Syk inhibitor with promising preclinical activity in diffuse large B-cell lymphoma. [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 4036.
Abstract 4037: Doxazosin exerts anti-metastatic potential in triple-negative breast cancer via impairment of cancer stem-like features
Authors: Kim Seongjae , Ko Dongmi , Seo Juyeon , Park Soeun , Park Minsu , Nam Kee Dal , koo Kang Yong , Seock So Ra , Park Jaeyoun , Oh Eunhye , Jung Eunsun , Kim Yoon-Jae , Kim Ji Young , Seo Jae Hong
Published: April 2023

Background and Purpose: Triple-negative breast cancer (TNBC) tumors typically harbor a high cancer stem-like population leading to chemo-resistance, recurrence, and metastasis. Tumor metastasis is associated with 90% of cancer-related deaths, highlighting the urgent clinical unmet need. Doxazosin is known to inhibit cell migration and invasion in several cancer cell types; however, the precise mechanisms underlying doxazosin’s anticancer effects in TNBC have not been fully elucidated. In the present study, we sought to investigate the mechanism of action of doxazosin responsible for its effects on apoptosis, cancer stem cell (CSC)-like properties, cell migration, and metastasis in TNBC.

Experimental designs: Doxazosin on TNBC cell lines [MDA-MB-231, BT549, and 4T1] in vitro was evaluated in cell viability, apoptosis, cell migration, CD44/CD24 staining, ALDH1 activity, and mammosphere formation. The effect of doxazosin on tumor growth, angiogenesis, and metastasis was evaluated in an orthotopic allograft mice model with CSC-enriched population.

Results: Doxazosin significantly reduced cell viability and induced apoptosis in MDA-MB-231 and BT549 cells via activation of caspase-3/-7 and cleavage of PARP. Doxazosin significantly suppressed cell migratory capability, concomitant with disrupting cytoskeletal proteins, including vimentin and F-actin expression in TNBC cells. An impairment of BCSC-like properties was associated with reduction of ALDH1 activity and the CD44+/CD24- population, concomitant with suppression of mammosphere-forming ability. Doxazosin administration reduced tumor growth and lung metastasis, as evidenced by a sharp decline in bioluminescence signal intensity. Inhibitory effect of tumor growth was accompanied by a significant decrease of Ki-67 and enhancement of apoptosis with DNA fragmentation and increased cleaved-caspase-3 expression. The latter phenomenon was associated with the impediment of JAK2/STAT3 signaling pathway and CSC-like properties. Furthermore, no toxic effects of doxazosin were found in liver and kidney function in animals.

Conclusion: Taken together, our findings highlight doxazosin as a promising candidate for drug repurposing in suppressing metastatic TNBC.

Citation Format: Seongjae Kim, Dongmi Ko, Juyeon Seo, Soeun Park, Minsu Park, Kee Dal Nam, Yong koo Kang, So Ra Seock, Jaeyoun Park, Eunhye Oh, Eunsun Jung, Yoon-Jae Kim, Ji Young Kim, Jae Hong Seo. Doxazosin exerts anti-metastatic potential in triple-negative breast cancer via impairment of cancer stem-like features. [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 4037.
Abstract 4106: Non-autonomous enhancement of gPDL1 CAR-T annihilates TNBC development
Authors: Li Chia-Wei , Wang Shih-Han , Lai Yun-Ju , Wang Jyun , Kuo Chun-Tse , Hung Shih-Duo Hsu , Liu Shou-Hou
Published: April 2023

Triple-negative breast cancer (TNBC) is the most aggressive and challenging breast cancer subtype, which does not respond to traditional endocrine and anti-HER2-targeted therapies. PD-L1 is highly enriched in TNBC and has been considered a therapeutic target. Despite the excellent anti-cancer activity, the atezolizumab-based chimeric antigen receptor (CAR) T cells showed a robust off-target effect. In addition, the treatment of solid tumors with CAR-T is limited by abnormal glycosylation in malignant tumors. Targeting glycosylated PD-L1 (gPD-L1) provided tissue specificity against TNBC, implying it can prevent antigen escape and off-target effect. In this study, we generated gPD-L1 CAR-T cells using lentiviral vectors expressing the scFv regions of the anti-gPD-L1 antibody. The gPD-L1 CAR-T cells exhibited antigen-specific activation, cytokine production, and cytolytic activity against TNBCs in vitro and in the xenograft tumors model. CyTOF and single-cell RNA sequencing (scRNA-seq) showed distinct IFNγ-positive cell types. Mechanistically, IFNγ crosstalked with EGFR signaling through Src activation and, in turn, triggered B3GNT3-mediated PD-L1 glycosylation. Inhibition of Src resulted in reduced gPD-L1 expression in TNBC. CRISPR/Cas9 knockout of B3GNT3 in TNBC cells impaired gPD-L1 CAR-T response. As a result of nonautonomous gPDL1 amplification in TNBCs, gPD-L1 CAR-T cells continued to annihilate TNBCs. Additionally, since gPD-L1 CAR-T cells provided higher specificity on TNBC, they had lower normal tissue toxicity. Overall, gPD-L1 CAR-T exhibits excellent anti-tumor activity against TNBCs, and it could be a promising immunotherapy tool to treat TNBCs in clinic. Furthermore, targeting glycosylation moiety on the tumor antigen is a novel approach to lessen CAR-T toxicity in patients.

Citation Format: Chia-Wei Li, Shih-Han Wang, Yun-Ju Lai, Jyun Wang, Chun-Tse Kuo, Shih-Duo Hsu Hung, Shou-Hou Liu. Non-autonomous enhancement of gPDL1 CAR-T annihilates TNBC development. [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 4106.
Abstract 4160: Characterization of cytotoxic resilient T cells in patients with advanced cancers in response to salvage radiation therapy or chemotherapy
Authors: Gicobi Joanina K. , Yan Yiyi , Zeng Hu , Park Sean , Dong Haidong
Published: April 2023

Although cancer burden in patients with advanced diseases results in T cell exhaustion and failed prior therapies, some of them achieved durable clinical responses to salvage therapy (chemotherapy or radiation therapy) implying a presence of tumor-reactive resilient T cells in them. However, the phenotype and functional features of resilient T cells have not been clearly defined. We found that CX3CR1+ CD8+ T cells with low mitochondrial membrane potential demonstrated features of resilient T cells: (a) their cytotoxic T cell function (degranulation/CD107a expression) recovered quickly after tumor burden has been reduced by radiation therapy or chemotherapy; (b) they maintain a low profile of glycolysis but produce optimal levels of ATP; (c) they express lower levels of PD-1 and TOX; (4) they share a feature of stem-like T cells (TCF1+PD-1+) that can be responsive to immunotherapy; (5) they avoid accumulation of a harmful level of cytosolic reactive oxygen species (ROS) that may cause T cell exhaustion; (6) they upregulate malic enzyme 1 (ME1) to balance glycolysis and ROS, and to expand cytotoxic effector T cells. These unique features of resilient T cells may not only be used as T cell biomarker to design rational combination therapy to control advanced cancers but can also be used to convey a resilient functional state to T cells in patients who lack them.

Citation Format: Joanina K. Gicobi, Yiyi Yan, Hu Zeng, Sean Park, Haidong Dong. Characterization of cytotoxic resilient T cells in patients with advanced cancers in response to salvage radiation therapy or chemotherapy. [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 4160.
Abstract 4162: T-cell metabolic profiling with bioluminescent assays
Authors: Bach Maggie , Karassina Natasha , Gilden Julia , Leippe Donna , Valley Mike , Sylvester Kayla , Vidugiriene Jolanta
Published: April 2023

Cellular energy metabolism is crucial for cell survival and the balance of metabolic pathways determines the health of the cell. Determining and understanding metabolic profiles of cells can aid in overall insight of cell health and improvement of cell therapeutics, especially for immune interventions. T-cells actively regulate their metabolic profile from quiescent to a highly metabolically active state after activation. Activation rewires the cell to drive glycolysis and oxidative phosphorylation for production of reducing power and ATP for cell growth and proliferation. A small population of these cells then transition to a more quiescent state and persist as memory cells to aid in secondary responses. Although the trend of T-cell metabolism after activation has been studied, the method of investigation is cumbersome and lacks depth of information of overall cell health. To address this, we developed a bioluminescence NAD(P)H core technology and applied it to build a set of cell-based assays covering diverse metabolic pathways including glucose, glycogen, and amino acid metabolism, and the TCA cycle. Here we utilize those assays to investigate changes in metabolic profiles of naïve, effector, and memory T-cells as they transition from a state of oxidative phosphorylation to increased glycolytic rates, dependence on different fuel consumption and storage, including an increase in fatty acid oxidation and glycogen accumulation. We demonstrate flexibility of the assays for detecting multiple metabolites from the same sample and monitoring metabolite changes over time by assaying multiple samples. We show that the assays are amenable to various volumes (96-,384-well plates) and low cell numbers. For example, activity of dehydrogenases involved in glycolysis, pentose phosphate and TCA cycles can be detected with less than 400 cells/well. In conclusion, understanding the processes governing dynamic metabolic changes of T-cells during their transition from naïve to activated to memory phenotypes is required for next-generation cell therapy development. The developed bioluminescence cell-based assays provide the sensitivity, throughput and robustness required for rapid evaluation of such changes.

Citation Format: Maggie Bach, Natasha Karassina, Julia Gilden, Donna Leippe, Mike Valley, Kayla Sylvester, Jolanta Vidugiriene. T-cell metabolic profiling with bioluminescent assays. [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 4162.
Abstract 4193: N-NOSE technology based on C. elegans olfaction. Multi-cancer early detection with high sensitivity using urine. Pancreatic-cancer-specific early detection with high sensitivity using urine
Authors: Luccio Eric Di , Hirotsu Takaaki
Published: April 2023

N-NOSE (Nematode Nose) is a cancer screening test based on the chemotactic characteristics of the nematode C. elegans. C. elegans shows avoidance of the urine of healthy individuals while displaying a chemotactic attraction toward the urine of patients with 15 types of cancer (stomach, colon-rectum, lung, breast, pancreas, liver, prostate, uterus, esophagus, gallbladder, bile duct, kidney, urinary bladder, ovary, oropharynx) rendering N-NOSE a primary multi-cancer screening test. N-NOSE has a high sensitivity of 87.5% and 90.2% specificity, on average, from early stage 0-I. N-NOSE is non-invasive because it relies on a urine sample and is affordable (JPY 13,800, ~USD 100).N-NOSE by Hirotsu Bio Science has been available in Japan since late 2020 and is a commercial hit with over 250,000 screenings performed, rapidly increasing. In this presentation, we outline the N-NOSE technology and business in Japan. We review N-NOSE clinical research and critical findings compared to competitors that highlight its excellent performance as a non-invasive primary cancer screening for early detection. N-NOSE is a multi-cancer early detection test. Next, we introduce an evolution of N-NOSE for identifying the cancer type. Pancreatic cancer is the 12th most common cancer worldwide but remains one the deadliest cancer with a 5-year relative survival rate at only 11%, all stages combined. An affordable, non-invasive, and highly sensitive early-stage pancreatic cancer screening test is sorely needed. As an evolution of N-NOSE we genetically engineered C. elegans by knocking out in AWC olfactory neurons a specific olfactory receptor (GPCR), triggering a chemotactic response particular to the only pancreatic cancer urine specimen at an early stage. In this study, we outline, for the first time, the critical discovery of cr-4 GPCR in AWC, which is crucial for a specific chemotaxis response to the urine of patients with pancreatic cancer. Next, we briefly conclude by outlining the link between N-NOSE primary cancer screening followed by N-NOSE cancer type identification, effectively completing the whole cancer screening and identification process in a non-invasive way.

Citation Format: Eric Di Luccio, Takaaki Hirotsu. N-NOSE technology based on C. elegans olfaction. Multi-cancer early detection with high sensitivity using urine. Pancreatic-cancer-specific early detection with high sensitivity using urine. [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 4193.
Abstract 42: In vivo subcutaneous and orthotopic cancer xenograft modeling in the SRG immunodeficient rat
Authors: Begemann Diane , Javidan Aida , Dunn Cynthia , Johnston Nicolas , Walton R. Grace , Steffey Valeriya , Corbin Ian , Fulcher Niveen , De Oliveira Cleusa , Xu Hu , Uzelac Mila , Deweyert Andrew , Ronald John A. , Schmid Susanne , Hebb Matthew O. , Noto Fallon K.
Published: April 2023

Human cancer xenografts are a vital tool for understanding tumor biology, growth kinetics, and therapeutic efficacy using animal models. Historically, immunodeficient mice have been the standard rodent species for cancer xenograft modeling. However, an immunodeficient rat that supports a wide variety of human cancer cell types would provide a larger rodent strain for easier surgical manipulation, serial blood sampling, and provide a single model in which efficacy, pharmacokinetics, and toxicology can be performed. We have created a Sprague Dawley Rag2 -/-, Il2rg -/- rat (SRGTM OncoRat®) that provides a highly supportive environment for growing tumors of human origin. The SRG rat lacks B, T, and NK cells and readily supports the growth of multiple human cancer cell lines. The SRG rat is more immunodeficient than the Nude rat, suggesting it may be permissive to a wider variety of human cancer types. Here we demonstrate the utility of the SRG rat for both subcutaneous and orthotopic xenograft modeling. The SRG rat supports the growth of both lung and liver orthotopic cancers. In addition, the SRG rat supports the growth of orthoptic human glioblastoma multiforme in the brain. We use in vivo imaging to visualize tumor establishment and growth in subcutaneous, orthotopic, and metastatic models. Furthermore, our data show the ability of the SRG rat to support the growth of multiple different human cancer cell types subcutaneously in two different matrices, Matrigel® and VitroGel®. These data confirm that the SRG rat is an excellent host for studying human cancer. Our data demonstrate that the SRG rat has a high utility for studies using in vivo imaging, orthotopic tumor implantation, and standard subcutaneous tumor modeling. As the most immunodeficient rat commercially available, the SRG rat supports the growth of multiple human cancer types in a larger rodent strain relative to commercially available mouse models.

Citation Format: Diane Begemann, Aida Javidan, Cynthia Dunn, Nicolas Johnston, R. Grace Walton, Valeriya Steffey, Ian Corbin, Niveen Fulcher, Cleusa De Oliveira, Hu Xu, Mila Uzelac, Andrew Deweyert, John A. Ronald, Susanne Schmid, Matthew O. Hebb, Fallon K. Noto. In vivo subcutaneous and orthotopic cancer xenograft modeling in the SRG immunodeficient rat [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 42.
Abstract 4246: Synergistic inhibition of prostate tumor growth and progression by the combination of curcumin and ursolic acid in HiMyc mice
Authors: Friedman Chelsea A. , Saha Achinto , DiGiovanni John
Published: April 2023

Prostate cancer (PCa) is the second leading cause of cancer death among men in the United States. According to the World Health Organization, approximately one-third of all cancer deaths could be prevented through an increased consumption of natural compounds. Through the use of a two-tiered screening method in PCa cells (depletion of cellular ATP level followed by glutamine uptake) of a phytochemical library, curcumin and ursolic acid were selected for further investigation. Twenty-four hour treatment of LNCaP, 22Rv1 and HMVP2 cells with the combination resulted in significant decreases in expression of members of the miR-17-92 and miR-106b-25 clusters, which are upregulated in PCa and have been shown to be involved in tumorigenesis, cell survival, and metastasis. In the HiMyc mouse model, where mice spontaneously develop primary PCa, supplementing diets with the combination of curcumin (1%) and ursolic acid (0.2%) resulted in significant inhibition of tumor growth and progression compared to diets containing either compound alone. There were no differences in body weight or food consumption in any of the diet groups during the tumor experiment. The current data suggests that the combination of curcumin and ursolic acid synergistically inhibits PCa growth and progression in HiMyc mice, which may be in part due to their effects on microRNA expression. Ongoing studies are further examining the role of specific miRNAs as well as oncogenic signaling pathways potentially involved in the mechanism of synergistic chemopreventive action of this combination. The current data suggest that this novel combination could be a valuable therapeutic option for prevention and/or treatment of PCa. Funding: This work was supported by NIH Grant CA228404.

Citation Format: Chelsea A. Friedman, Achinto Saha, John DiGiovanni. Synergistic inhibition of prostate tumor growth and progression by the combination of curcumin and ursolic acid in HiMyc mice. [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 4246.
Abstract 4252: CCNE1 amplification in high-grade serous ovarian cancer: analysis of the GENIE database
Authors: Al-Akhras Ashaar , Tashtoush Shaden , Ghazou Alina
Published: April 2023

Background: High-grade serous ovarian cancer (HGSOC) is the most common histological subtype of ovarian cancer as well as the most lethal. A primary oncogenic driver in a subset of HGSOC patients, CCNE1 amplification is associated with poorer outcomes and resistance to standard treatment. This study aims to investigate the copy number gain of CCNE1 and the genetic mutations associated with CCNE1 amplification using a large cohort of clinico-genomic data. Methods: We queried the American Association of Cancer Research-Genomics Evidence Neoplasia Information Exchange (GENIE) registry database v12.1 for patients with HGSOC. We studied CCNE1 amplification and the clinical, demographic, and genetic mutation associations. Results: A total of 3297 HGSOC samples were extracted from the registry. The mean age of patients at sequencing was 62 ± 10 years. Out of 2416 samples profiled for copy number alterations, CCNE1 amplification was found in 13.6%, being the most commonly amplified gene, followed by AGO2 (11.7%) and MYC (9.7%). Co-amplification between CCNE1 and AKT2 was demonstrated. Between the 397 samples with CCNE1 amplification and 2088 without, the genes that were more frequently mutated in the CCNE1 amplified group were AKT2 (20.4% vs 3.2%, p<0.001), KRAS (11.3% vs 4.3%, p<0.001), BRD4 (8.0% vs 4.4%, p = 0.006), and ERBB2 (7.9% vs 4.0%, p = 0.003). Other genes were less frequently mutated in the CCNE1 amplified cohort such as NF1 (7.6 % vs 12.9%, p = 0.009), BRCA1 (3.7% vs 10.0%, p<0.001), BRCA2 (2.7% vs 7.4%, p = 0.003), RB1 (2.7% vs 6.4%, p = 0.013), and PTEN (1.2% vs 4.0%, p = 0.02). Frequencies of MYC, PIK3CA, KMT2D, CDK12, and TSC2 mutations were similar between the two groups. TP53 was mutated in 100% of samples. There were no significant differences between the two groups in primary race or total mutation count, and mean age at sequencing was similar. Conclusion: Somatic sequencing is warranted to identify patients of this subset of HGSOC, and understanding the co-amplifications and co-mutations is essential so that they are targeted for optimal survival outcomes.

Citation Format: Ashaar Al-Akhras, Shaden Tashtoush, Alina Ghazou. CCNE1 amplification in high-grade serous ovarian cancer: analysis of the GENIE database. [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 4252.