bluuebunny/crossref_metadata_2025 huggingface dataset:
Data from Functional Analysis of Somatic Mutations Affecting Receptor Tyrosine Kinase Family in Metastatic Colorectal Cancer
Authors: Duplaquet Leslie , Figeac Martin , Leprêtre Frédéric , Frandemiche Charline , Villenet Céline , Sebda Shéhérazade , Sarafan-Vasseur Nasrin , Bénozène Mélanie , Vinchent Audrey , Goormachtigh Gautier , Wicquart Laurence , Rousseau Nathalie , Beaussire Ludivine , Truant Stéphanie , Michel Pierre , Sabourin Jean-Christophe , Galateau-Sallé Françoise , Copin Marie-Christine , Zalcman Gérard , De Launoit Yvan , Fafeur Véronique , Tulasne David
Published: April 2023

Abstract
Besides the detection of somatic receptor tyrosine kinases (RTK) mutations in tumor samples, the current challenge is to interpret their biological relevance to give patients effective targeted treatment. By high-throughput sequencing of the 58 RTK exons of healthy tissues, colorectal tumors, and hepatic metastases from 30 patients, 38 different somatic mutations in RTKs were identified. The mutations in the kinase domains and present in both tumors and metastases were reconstituted to perform an unbiased functional study. Among eight variants found in seven RTKs (EPHA4-Met726Ile, EPHB2-Val621Ile, ERBB4-Thr731Met, FGFR4-Ala585Thr, VEGFR3-Leu1014Phe, KIT-Pro875Leu, TRKB-Leu584Val, and NTRK2-Lys618Thr), none displayed significantly increased tyrosine kinase activity. Consistently, none of them induced transformation of NIH3T3 fibroblasts. On the contrary, two RTK variants (FGFR4-Ala585Thr and FLT4-Leu1014Phe) caused drastic inhibition of their kinase activity. These findings indicate that these RTK variants are not suitable targets and highlight the importance of functional studies to validate RTK mutations as potential therapeutic targets.
Data from A Novel Anti-HER2 Antibody–Drug Conjugate XMT-1522 for HER2-Positive Breast and Gastric Cancers Resistant to Trastuzumab Emtansine
Authors: Le Joncour Vadim , Martins Ana , Puhka Maija , Isola Jorma , Salmikangas Marko , Laakkonen Pirjo , Joensuu Heikki , Barok Mark
Published: April 2023

Abstract
Most patients with HER2-positive breast or gastric cancer exhibit primary or acquired resistance to trastuzumab emtansine (T-DM1), and such patients may have limited therapeutic options. XMT-1522 is a novel anti-HER2 antibody–drug conjugate. We compared XMT-1522 to T-DM1 in preclinical models. The effects of XMT-1522 and T-DM1 on cell survival and apoptosis were compared in six HER2-positive breast cancer or gastric cancer cell lines, of which three lines were T-DM1–sensitive (N-87, OE-19, JIMT-1) and three T-DM1–resistant (RN-87, ROE-19, SNU-216). We compared these agents also in the HER2-negative breast cancer cell line MCF-7, and in mouse RN-87 and JIMT-1 xenograft models. Cell survival was assessed using the AlamarBlue method and apoptosis with the Caspase-Glo 3/7 method. XMT-1522 inhibited the growth of all six HER2-positive cell lines. The proportions of cells that survived XMT-1522 treatment were smaller as compared with T-DM1, particularly in the T-DM1–resistant cell lines. XMT-1522 induced more cell apoptosis compared with T-DM1. While RN-87 and JIMT-1 xenograft tumors progressed on T-DM1 treatment, all tumors responded to XMT-1522, and all but one tumor disappeared during the XMT-1522 treatment. XMT-1522 had a strong antitumor effect on RN-87 and JIMT-1 xenografts that progressed on T-DM1. We conclude that XMT-1522 was effective in HER2-positive breast cancer and gastric cancer cell lines resistant to T-DM1, and in xenograft models resistant to T-DM1. The results support the testing of XMT-1522 in clinical trials in patients with HER2-positive cancer.
Data from A Novel Therapeutic Strategy for Pancreatic Cancer: Targeting Cell Surface Glycan Using rBC2LC-N Lectin–Drug Conjugate (LDC)
Authors: Shimomura Osamu , Oda Tatsuya , Tateno Hiroaki , Ozawa Yusuke , Kimura Sota , Sakashita Shingo , Noguchi Masayuki , Hirabayashi Jun , Asashima Makoto , Ohkohchi Nobuhiro
Published: April 2023

Abstract
Various cancers, including pancreatic ductal adenocarcinoma (PDAC), remain intractable even with costly tumor-targeting antibody drugs. Because the outermost coatings of cancer cells are composed of cell-specific glycan layers (glycocalyx), lectins, proteins with glycan-binding potential, were evaluated for possible use as drug carriers in PDAC treatment. A human PDAC cell line with well-to-moderately differentiated properties (Capan-1) was subjected to lectin microarray analysis to identify specific lectin–glycan pairs. The selected lectin was fused with a bacterial exotoxin for the construction of a lectin–drug conjugate (LDC), and its safety and antitumor effects were evaluated. A specific affinity between a recombinant bacterial C-type lectin (rBC2LC-N) and Capan-1 was identified, and its positivity was confirmed in 69 human samples. In contrast to the belief that all lectins mediate harmful hemagglutination, rBC2LC-N did not cause hemagglutination with human erythrocytes and was safely administered to mice. The 50% inhibitory concentration of LDC to Capan-1 (1.04 pg/mL = 0.0195 pmol/L) was 1/1,000 lower than that reported for conventional immunotoxins. The intraperitoneal administration of LDC reduced the tumor weight from 390 to 130.8 mg (P < 0.01) in an orthotopic model and reduced the number of nodules from 48 to 3 (P < 0.001) and improved survival from 62 to 105 days in a peritoneal dissemination model (P < 0.0001). In addition, the effect of LDC was reproduced in nodules from patient-derived PDAC xenografts through intravenous injection. Herein, we show the concept of utilizing lectins as drug carriers to target glycans on the cancer cell surface, highlighting new insights into cancer treatments. Mol Cancer Ther; 17(1); 183–95. ©2017 AACR.
Data from Comparative Oncology Evaluation of Intravenous Recombinant Oncolytic Vesicular Stomatitis Virus Therapy in Spontaneous Canine Cancer
Authors: Naik Shruthi , Galyon Gina D. , Jenks Nathan J. , Steele Michael B. , Miller Amber C. , Allstadt Sara D. , Suksanpaisan Lukkana , Peng Kah Whye , Federspiel Mark J. , Russell Stephen J. , LeBlanc Amy K.
Published: April 2023

Abstract
Clinical translation of intravenous therapies to treat disseminated or metastatic cancer is imperative. Comparative oncology, the evaluation of novel cancer therapies in animals with spontaneous cancer, can be utilized to inform and accelerate clinical translation. Preclinical murine studies demonstrate that single-shot systemic therapy with a vesicular stomatitis virus (VSV)-IFNβ-NIS, a novel recombinant oncolytic VSV, can induce curative remission in tumor-bearing mice. Clinical translation of VSV-IFNβ-NIS therapy is dependent on comprehensive assessment of clinical toxicities, virus shedding, pharmacokinetics, and efficacy in clinically relevant models. Dogs spontaneously develop cancer with comparable etiology, clinical progression, and response to therapy as human malignancies. A comparative oncology study was carried out to investigate feasibility and tolerability of intravenous oncolytic VSV-IFNβ-NIS therapy in pet dogs with spontaneous cancer. Nine dogs with various malignancies were treated with a single intravenous dose of VSV-IFNβ-NIS. Two dogs with high-grade peripheral T-cell lymphoma had rapid but transient remission of disseminated disease and transient hepatotoxicity that resolved spontaneously. There was no shedding of infectious virus. Correlative pharmacokinetic studies revealed elevated levels of VSV RNA in blood in dogs with measurable disease remission. This is the first evaluation of intravenous oncolytic virus therapy for spontaneous canine cancer, demonstrating that VSV-IFNβ-NIS is well-tolerated and safe in dogs with advanced or metastatic disease. This approach has informed clinical translation, including dose and target indication selection, leading to a clinical investigation of intravenous VSV-IFNβ-NIS therapy, and provided preliminary evidence of clinical efficacy and potential biomarkers that correlate with therapeutic response. Mol Cancer Ther; 17(1); 316–26. ©2017 AACR.
Data from TAS4464, A Highly Potent and Selective Inhibitor of NEDD8-Activating Enzyme, Suppresses Neddylation and Shows Antitumor Activity in Diverse Cancer Models
Authors: Yoshimura Chihoko , Muraoka Hiromi , Ochiiwa Hiroaki , Tsuji Shingo , Hashimoto Akihiro , Kazuno Hiromi , Nakagawa Fumio , Komiya Yu , Suzuki Satoshi , Takenaka Toru , Kumazaki Masafumi , Fujita Naoya , Mizutani Takashi , Ohkubo Shuichi
Published: April 2023

Abstract
NEDD8-activating enzyme (NAE) is an essential E1 enzyme of the NEDD8 conjugation (neddylation) pathway, which controls cancer cell growth and survival through activation of cullin-RING ubiquitin ligase complexes (CRL). In this study, we describe the preclinical profile of a novel, highly potent, and selective NAE inhibitor, TAS4464. TAS4464 selectively inhibited NAE relative to the other E1s UAE and SAE. TAS4464 treatment inhibited cullin neddylation and subsequently induced the accumulation of CRL substrates such as CDT1, p27, and phosphorylated IκBα in human cancer cell lines. TAS4464 showed greater inhibitory effects than those of the known NAE inhibitor MLN4924 both in enzyme assay and in cells. Cytotoxicity profiling revealed that TAS4464 is highly potent with widespread antiproliferative activity not only for cancer cell lines, but also patient-derived tumor cells. TAS4464 showed prolonged target inhibition in human tumor xenograft mouse models; weekly or twice a week TAS4464 administration led to prominent antitumor activity in multiple human tumor xenograft mouse models including both hematologic and solid tumors without marked weight loss. As a conclusion, TAS4464 is the most potent and highly selective NAE inhibitor reported to date, showing superior antitumor activity with prolonged target inhibition. It is, therefore, a promising agent for the treatment of a variety of tumors including both hematologic and solid tumors. These results support the clinical evaluation of TAS4464 in hematologic and solid tumors.
Data from Precision Chemoradiotherapy for HER2 Tumors Using Antibody Conjugates of an Auristatin Derivative with Reduced Cell Permeability
Authors: Hingorani Dina V. , Doan Matthew K. , Camargo Maria F. , Aguilera Joseph , Song Seung M. , Pizzo Donald , Scanderbeg Daniel J. , Cohen Ezra E.W. , Lowy Andrew M. , Adams Stephen R. , Advani Sunil J.
Published: April 2023

Abstract
The most successful therapeutic strategies for locally advanced cancers continue to combine decades-old classical radiosensitizing chemotherapies with radiotherapy. Molecular targeted radiosensitizers offer the potential to improve the therapeutic ratio by increasing tumor-specific kill while minimizing drug delivery and toxicity to surrounding normal tissue. Auristatins are a potent class of anti-tubulins that sensitize cells to ionizing radiation damage and are chemically amenable to antibody conjugation. To achieve tumor-selective radiosensitization, we synthesized and tested anti-HER2 antibody–drug conjugates of two auristatin derivatives with ionizing radiation. Monomethyl auristatin E (MMAE) and monomethyl auristatin F (MMAF) were attached to the anti-HER2 antibodies trastuzumab and pertuzumab through a cleavable linker. While MMAE is cell permeable, MMAF has limited cell permeability as free drug resulting in diminished cytotoxicity and radiosensitization. However, when attached to trastuzumab or pertuzumab, MMAF was as efficacious as MMAE in blocking HER2-expressing tumor cells in G2–M. Moreover, MMAF anti-HER2 conjugates selectively killed and radiosensitized HER2-rich tumor cells. Importantly, when conjugated to targeting antibody, MMAF had the advantage of decreased bystander and off-target effects compared with MMAE. In murine xenograft models, MMAF anti-HER2 antibody conjugates had less drug accumulated in the normal tissue surrounding tumors compared with MMAE. Therapeutically, systemically injected MMAF anti-HER2 conjugates combined with focal ionizing radiation increased tumor control and improved survival of mice with HER2-rich tumor xenografts. In summary, our results demonstrate the potential of cell-impermeable radiosensitizing warheads to improve the therapeutic ratio of radiotherapy by leveraging antibody–drug conjugate technology.
Abstract 1016: Prognostic factors for overall survival, recurrence-free survival, and metastasis-free survival of colon cancer in Morocco
Authors: Agy Fatima El , Chbani Laila
Published: April 2023

Introduction: Colon cancer is the third most common cancer in Morocco. Its incidence and mortality rate is constantly increasing in this country. This requires the determination of clinical and molecular prognostic factors of CC in our context. Studies indicate that mutations in the RAS genes (KRAS and NRAS) represent such a factor. The aim of this study is to evaluate the prognostic impact of KRAS and NRAS mutations on overall survival (OS), recurrence-free survival (RFS) and metastasis-free survival (MFS) of CC patients.

Methods This study involved a series of 210 patients with colon cancer. The clinical and pathological data were obtained from of each patient’s files at the molecular pathology department of the CHU HASSAN II of FES. For molecular data, full RAS status was determined by direct sequencing and Q24 pyrosequencing, after extraction of DNA from formalin-fixed and paraffin-embedded tissue. The evaluation of the relationship between OS, RFS, and MFS and the molecular and clinical factors of the patients was performed by the Kaplan-Meier method and the log-rank test.

Results The average age of our patients was 57.5 ± 14.6 years. A slight male predominance was found (52.7%). 39.5% of the cases had a mutation in one of the RAS genes. Mutation of exon 2 of the KRAS gene was the most frequent with a percentage of 36.6%. The NRAS gene was mutated in 2.8% of our patients. The exon 2 mutated tumors were significantly associated with female gender (p=0.04), and left location (p=0.04) compared to the other exons. The mean OS, RFS, and MFS were 100.0%, 92.0% and 92.1% respectively. OS was significantly longer in tumors with KRAS exon 4 mutations compared to tumors with other types of RAS mutations and was also associated with right colon tumors (P=0.001), localized stages I-II (P=0.001), tumors without metastatic nodes (P=0.03), and MSI tumors (P=0.01). For RFS and MFS, localized stage and tumors without metastatic nodes were good prognostic factors (p=0.002, p=0.000). However, the type of mutated exon had no prognostic impact on either.

Conclusion In our setting right location, localized stage, absence of metastatic nodes, MSI status and mutated status of exon 4 of the KRAS gene (compared to other exons) are all predictive factors of better overall survival of colon cancer in Morocco. The type of the mutated exon of the RAS genes has no impact on RFS and MFS.

Citation Format: Fatima El Agy, Laila Chbani. Prognostic factors for overall survival, recurrence-free survival, and metastasis-free survival of colon cancer in Morocco [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 1016.
Abstract 1020: Tissue-informed ctDNA MRD assay detects post-surgery minimal residual disease in HCC patients
Authors: Hu Jie , Tang Haoran , Xie Feng , Zhang Yue , Jia Shidong , Zhou Jian
Published: April 2023

Introduction: Residual disease detection is a significant clinical need in post-surgery treatment of resectable hepatocellular carcinoma (HCC) patients. Previous NGS-based studies targeting fixed genomic regions had reported that circulating tumor DNA (ctDNA) could provide evidence for prognosis prediction during HCC treatment. However, the sensitivity of methods was limited and could not reach the threshold of minimal residual disease (MRD) test. Here we reported a baseline tissue -informed ctDNA NGS assay, detecting MRD in HCC patients who received hepatectomy and predicted prognosis in MRD positive patients.

Methods: The study retrospectively enrolled 200 early-stage HCC patients who received surgical resections. Tumor and normal tissue samples were collected respectively through surgery. Plasma samples were collected 7 days or more post-surgery. In the completed pilot study, samples from 17 patients were tested by a baseline tissue-informed MRD assay (PredicineBEACON). Tumor-specific mutations were identified by whole exon sequencing on baseline tumor and normal tissue samples. For each patient, up to 50 mutations were selected through bioinformatics pipelines for personalized MRD panel design, in combination with a fixed panel of 500 tumor actionable hotspots. The post-surgery plasma samples were then tested for MRD with the panel through ultra-deep sequencing(100,000X). Afterward, genomic tumor fraction proportions were calculated.

Results: In the completed pilot study, 12 of 17 patients (70.6%) were called MRD positive based on tests of post-surgery plasma. Among MRD positive patients, 3 were identified as MRD risk high (proportion of tumor fraction≥0.1%) and MRD risk of others were moderate (proportion of tumor fraction<0.1%). The tumor fraction proportion of MRD positive patients varied from 0.013% to 17.84% (median 0.042%). The recurrence free survival (RFS) and overall survival (OS) probabilities were significantly correlated with MRD risk: the median RFS of two groups(risk-high vs. risk-moderate) was 10.1 months vs. 57.4 months (p-value<0.001), while the median OS of two groups (risk-high vs. risk-moderate) was 31.2 months vs. 57.4 months(p-value=0.028).

Conclusions: Baseline-informed ctDNA NGS assay showed ultra-sensitive capability of MRD detection on early-stage HCC patients, with outstanding positive rate through plasma samples collected 7 days post-surgery. The MRD risk provided significant prognostic evidence for patient survival and disease relapse.

Citation Format: Jie Hu, Haoran Tang, Feng Xie, Yue Zhang, Shidong Jia, Jian Zhou. Tissue-informed ctDNA MRD assay detects post-surgery minimal residual disease in HCC 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 1020.
Abstract 103: Dissecting cell state diversity during melanoma growth and metastasis
Authors: Karras Panagiotis , Bordeu Ignacio , Pozniak Joanna , Nowosad Ada , Pazzi Cecilia , Van Raemdonck Nina , Khoo Jia Hui , Pedri Dennis , Rustgi Anil , Bechter Oliver , Blanpain Cedric , Simons Benjamin , Rambow Florian , Marine Jean Christophe
Published: April 2023

Cellular plasticity endows cancer cells with capacity towards a phenotypic state. Except from the genetic origin of cellular plasticity, cancer cells can transit to reversible phenotypic states as a result of microenvironmental cues and are often driven by stochastic epigenetic and/or transcriptional fluctuations. Melanoma is a prime example of heterogeneous and plastic tumors where the origin and magnitude of cell state diversity remains poorly understood. Combining lineage tracing, 3D imaging and quantitative mathematical modelling in a clinically-relevant mouse model of melanoma, we demonstrated that tumors follow a hierarchical model of growth supported by a population of Melanoma Stem-like Cells (MSCs) that exhibit a transcriptomic signature of pre-migratory neural crest cells established transiently during embryonic development. ScRNA seq in mouse and drug naïve human biopsies revealed that MSCs are evolutionarily conserved and independent of the “genetic makeup” of the tumors. By deploying spatial transcriptomic approaches and multiplex imaging, we developed a spatially and temporally resolved map of the diversity of melanoma showing a non-random tumor organization. Multimodal analysis unraveled unique cell type and state interactions and importantly demonstrated that MSCs reside in perivascular niches favoring tumor growth. Endothelial Cells were found to have a key role in the acquisition of stemness properties. Co-culture assays led to melanoma dedifferentiation and proliferation advantage. Of note, supplementing melanoma cells with ECs accelerated the growth of tumors in vivo, where MSCs pool was increased. Consistent with a model in which only a fraction of cells are fated to fuel growth, temporal single-cell tracing of a population of melanoma cells harboring a mesenchymal-like state revealed that these cells do not contribute to primary tumor growth but, instead, constitutes a pool of metastatic-initiating cells that switch cell identity while disseminating to secondary organs. Taken together, these results will pave the way for the development of strategies that exploit the “chameleonic” nature of cancer cells and, ultimately, target MSC niche-dependent specification mechanisms.

Citation Format: Panagiotis Karras, Ignacio Bordeu, Joanna Pozniak, Ada Nowosad, Cecilia Pazzi, Nina Van Raemdonck, Jia Hui Khoo, Dennis Pedri, Anil Rustgi, Oliver Bechter, Cedric Blanpain, Benjamin Simons, Florian Rambow, Jean Christophe Marine. Dissecting cell state diversity during melanoma growth and metastasis [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 103.
Abstract 1070: The anti-tumor efficacy of combining oral ATR kinase inhibitor ceralasertib with TIC10/ONC201, an oral Akt/ERK inhibitor, TRAIL pathway and integrated stress response inducer, in prostate cancer treatment
Authors: Xia Yutong , Schwermann Maximilian P. , George Andrew , Ochsner Anna , Carneiro Benedito A. , El-Deiry Wafik S.
Published: April 2023

Prostate cancer is the most common cancer and the second leading cause of cancer death among men in the United States. There are 268,490 estimated new cases and 34,500 estimated deaths in 2022 according to the NIH. Patients with metastatic castration resistant prostate cancer (mCRPC) have a poor prognosis and the current treatment options show limited efficacy. From the TRAP trial, 20% of mCRPC patients are reported to bear DNA repair defects. Ceralasertib, formerly known as AZD6738, is a potent and selective orally bioavailable inhibitor of the ataxia tenlangiectasia and Rad3-related (ATR) kinase, which is involved in DNA repair in response to DNA damage and replication stress. Ceralasertib’s antitumor activity as a monotherapy in treating prostate cancer is moderate. Thus, we investigated a combination therapy of ceralasertib with ONC201, a potent and cytotoxic orally bioavailable inhibitor of the serine/threonine protein kinase Akt and extracellular signal-regulated kinase (ERK). Upon administration, ONC201 can induce tumor cell apoptosis mediated by tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and activate integrated stress response (IRS). Preliminary results demonstrate synergistic activity with the combination therapy in vitro using CellTiterGLO viability assay 72 hours in a 96 well plate post treatment. Ongoing studies using western blotting and cytokine profiling are intended to illustrate the mechanism behind the proven synergy. Our results aim to solve the overarching need in developing novel therapeutic strategies to overcome resistance in current prostate cancer treatments.

Citation Format: Yutong Xia, Maximilian P. Schwermann, Andrew George, Anna Ochsner, Benedito A. Carneiro, Wafik S. El-Deiry. The anti-tumor efficacy of combining oral ATR kinase inhibitor ceralasertib with TIC10/ONC201, an oral Akt/ERK inhibitor, TRAIL pathway and integrated stress response inducer, in prostate cancer treatment [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 1070.
Abstract 1099: Combined treatment with direct hydrogen peroxide administration and irradiation promotes an abscopal effect in mouse models
Authors: Kemmotsu Naoya , zhu Li , Ueda Youki , Dansako Hiromichi , Toyooka Shinichi , Nagasaki Joji , Togashi Yosuke
Published: April 2023

Background: Hydrogen peroxide (H2O2) causes oxidative stress and cytotoxicity and may be used in cancer therapy in combination with radiotherapy (RT). Indeed, H2O2 has been clinically developed as a radiosensitizer (development code: KRC-01). However, effects of H2O2 on antitumor immunity remain unclear. In this study, we investigated the effects, especially an abscopal effect, using mouse models.

Material and Method: We used MC-38 (mouse colon cancer) and B16F10 (mouse melanoma) cell lines. Tumor cells were implanted simultaneously in two locations; hind legs and back of mice. A self-made radiation shielding device was used to prevent irradiation of the tumors on the back. Only the hind legs were directly injected with a mixture of H2O2 and hyaluronic acid (KRC-01), and then irradiated. Immune cells in tumors or tumor draining lymph nodes were analyzed with flow cytometry.

Result: We compared tumor growth among four groups: control, irradiation alone, H2O2 alone, and combination groups. Compared to the other groups, the combination significantly reduced non-irradiated tumor growth (abscopal effect). In immunodeficient SCID mice, this effect was not observed. Flow cytometric analyses of tumor draining lymph nodes showed that dendritic cells were maturated in the combination groups compared to the others. In addition, CD8+ T cells were activated in the non-irradiated tumors from the combination group compared to the others.

Conclusion: Intratumoral H2O2 administration enhanced the abscopal effect through activating antitumor immunity by RT.

Citation Format: Naoya Kemmotsu, Li zhu, Youki Ueda, Hiromichi Dansako, Shinichi Toyooka, Joji Nagasaki, Yosuke Togashi. Combined treatment with direct hydrogen peroxide administration and irradiation promotes an abscopal effect in mouse 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 1099.
Abstract 112: Multi-omics analysis of molecular landscape and heterogeneity in fetal adenocarcinoma of the lung
Authors: Guo Wei , Sun Li , Chen Xiaoxi , Zhou Haitao , Yan Shi , Zhao Gang , Tang Haimeng , Bao Hua , Wu Xue , Shao Yang , Lin Lin
Published: April 2023

Background: Fetal adenocarcinoma of the lung (FLAC) is a rare form of lung adenocarcinoma and was divided into high-grade (H-FLAC) and low-grade (L-FLAC) subtypes. Until now, much of the knowledge regarding FLAC is from case reports or small case series and there was no comprehensive multi-omics study of FLAC.

Methods: A total of 20 FLAC patients who underwent tumor resection were retrospectively recruited in this study. Three regions from each of the 20 tumor specimens and matched normal tissue specimens were collected. Whole-exome sequencing, targeted bisulfite sequencing and whole transcriptome sequencing were performed on multi-regional tumor samples and normal tissue samples.

Results: We compared our FLAC cohort with a conventional East Asian-ancestry lung adenocarcinoma (LUAD) and revealed lower frequencies of altered driver genes on both somatic mutations and CNV levels, including EGFR, KRAS and ERRB2, in FLAC. Mutational signature analysis showed an enrichment of age-related mutational signatures in FLAC and tobacco-associated and defective DNA mismatch repair-related signatures were more enriched in H-FLAC and in L-FLAC, respectively. Significant differences between H-FLAC and L-FLAC were observed in the genomic and epigenomic landscape. In contrast, the transcriptomic landscape of H-FLAC closely dissembled L-FLAC by pathway enrichment analysis of differentially expressed genes. Furthermore, although genetic and epigenetic changes showed congruent evolutionary trajectories, the relationships between multi-level intratumor heterogeneity (ITH) were distinct in H-FLAC and L-FLAC. The genetic and transcriptional ITH showed a significant negative correlation in H-FLAC but a significant positive correlation in L-FLAC. Moreover, the lower genetic ITH was significantly related to a worse recurrence-free survival and overall survival in FLAC patients whereas lower methylation ITH H-FLAC patients tended to have better survival.

Conclusions: In this study, we first depicted the multi-omics landscapes of this rare lung cancer type and compared the two subtypes in a multi-omics and heterogeneity aspect. Different levels of ITH showed different relationships with survival, and non-genetic ITH, instead of genetic ITH, may contribute to the prognosis in FLAC patients.

Citation Format: Wei Guo, Li Sun, Xiaoxi Chen, Haitao Zhou, Shi Yan, Gang Zhao, Haimeng Tang, Hua Bao, Xue Wu, Yang Shao, Lin Lin. Multi-omics analysis of molecular landscape and heterogeneity in fetal adenocarcinoma of the lung [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 112.
Abstract 1138: Novel SMARCA2 degrading bifunctional molecules as therapeutics in SMARCA4 mutant lung cancer
Authors: Kotagiri Sasi , Blazanin Nicholas , Xi Yuanxin , Wang Jing , Lissanu Yonathan
Published: April 2023

Lung cancer is the top cause of cancer mortality. Despite recent advances, the majority of patients with lung cancer still lack effective therapeutic options, underscoring the dire need for additional treatment approaches. Genomic studies have identified frequent mutations in subunits of the SWI/SNF chromatin remodeling complex including SMARCA4 and ARID1A in non-small cell lung cancer with a frequency of up to 33% in advanced stage disease, making it the most frequently mutated complex in lung cancer. Recent reports, and our own data, have identified the paralogue SMARCA2 to be synthetic lethal to SMARCA4 suggesting SMARCA2 is a valuable therapeutic target. However, the discovery of selective inhibitors of SMARCA2 has been challenging. To overcome this hurdle, we have utilized iterative structure-activity relationship (SAR) studies to develop novel, potent and selective SMARCA2 degrading small molecules based on proteolysis targeting chimera (PROTAC) technology. We demonstrated that YD23, our lead SMARCA2 PROTAC, potently and selectively induces degradation of SMARCA2. Using global proteomic analysis and quantification of more than 8000 proteins, we showed that our SMARCA2 PROTAC is highly selective for SMARCA2. Importantly, we showed that YD23 selectively inhibits growth of SMARCA4 mutant lung cancer cells in vitro. Mechanistically, we demonstrated that YD23 reduces chromatin accessibility only in SMARCA4 deficient cells. In particular, YD23 profoundly decreased chromatin accessibility at enhancers of a number of genes including cell cycle and cell growth regulatory genes. Gene expression profiling and pathway analysis indicated that various cell cycle genes were downregulated by YD23 consistent with the reduced chromatin accessibility at their regulatory regions. In conclusion, our study provides a potent chemical probe for studying the synthetic lethal interaction between SMARCA2 and SMARCA4, dissect the chromatin and epigenetic landscape alterations and lay the foundation for future preclinical and clinical development of SMARCA2 degraders as therapeutics.

Citation Format: Sasi Kotagiri, Nicholas Blazanin, Yuanxin Xi, Jing Wang, Yonathan Lissanu. Novel SMARCA2 degrading bifunctional molecules as therapeutics in SMARCA4 mutant lung 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 1138.
Abstract 1160: A machine learning based method for in-vivo metabolic flux analysis of patient tumors
Authors: Mittal Anjali , Scott Andrew , Achreja Abhinav , Tarnal Vijay , Al-Holou Wajd , Lyssiotis Costas , Wahl Daniel , Nagrath Deepak
Published: April 2023

Metabolic fluxes likely control cancer phenotypes and treatment responses but cannot be measured in cancers in human patients. Machine learning has become a popular tool to study cancer biology and can identify complex patterns in the data which may not be decipherable by traditional analyses. We propose an approach to combine 13C-tracer analysis with machine learning to quantify the metabolism of purines in glioblastoma patients. In recent years, multiple studies have been conducted on isotope tracing in cancer patients. While the data for the enrichment of metabolites in human tumors has improved our understanding of cancer metabolism, we lack methods to estimate metabolic fluxes from these enrichment values. A significant challenge in estimating the fluxes in human subjects is that we are limited to enrichment data from a single in-vivo measurement at isotopic non-steady state. We overcome this challenge by combining INST-MFA (Isotopic Non-Stationary-Metabolic Flux Analysis) with machine learning methods. We use the enrichment of circulating metabolites as an input and simulate the time-dependent metabolite enrichment profiles for randomized flux vectors. We use this data to train a convolutional neural network (CNN) model to predict the flux ratios. Apart from being able to predict the fluxes from a single time-point, a machine learning model drastically reduces the time required to fit a traditional INST-MFA model. We demonstrate the real-world validity of the model by predicting flux ratios in mice models and comparing them to results from traditional INST-MFA models. We apply the model to decipher the heterogeneity of the purine pathway in eight patients with brain tumors.

Citation Format: Anjali Mittal, Andrew Scott, Abhinav Achreja, Vijay Tarnal, Wajd Al-Holou, Costas Lyssiotis, Daniel Wahl, Deepak Nagrath. A machine learning based method for in-vivo metabolic flux analysis of patient tumors [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 1160.
Abstract 125: Archival single cell sequencing reveals persistent subclones over years to decades of DCIS progression
Authors: Wang Kaile , Kumar Tapsi , wang Junke , Minussi Darlan , Sei Emi , li Jianzhuo , Tran Tuan , Thennavan Aatish , Hu Min , Casasent Anna , Xiao Zhenna , Bai Shanshan , Zhao Yuehui , Zurita Amado , Aparicio Ana , Chapin Brian , ye Jie , Zhang Jianjun , Gibbons Don , Futreal Andrew , King Lorraine , Marks Jeffrey , Hwang E. Shelley , Shah Vandna , Sawyer Ellinor , Kristel Petra , Wesseling Jelle , Lips Esther H. , Navin Nicholas
Published: April 2023

Ductal carcinoma in situ (DCIS) is a common precursor of invasive breast cancer (IBC), yet the genomic progression to recurrent disease remains poorly understood. A main contributor to this gap in knowledge arises from technical challenges with genomic profiling of formalin-fixed paraffin-embedded (FFPE) materials. To address this challenge, we developed Arc-well, the first high-throughput method that can perform single cell DNA sequencing of thousands of cells from FFPE materials and frozen tissues. Using Arc-well, we profiled genomic copy number in 27,851 single cells from 26 archival FFPE tissues that were stored for 3-31 years. Analysis of genomic evolution in 10 patients with matched DCIS and recurrent cancers (DCIS or IBC) separated by 2-16 years showed that many primary DCIS lesions had already undergone whole-genome-doubling and had extensive clonal diversity, similar to the paired recurrence. The data from most patients (8/10) suggest an evolutionary bottleneck model of progression, in which a single subclone persisted during the progression to the recurrent disease, revealing copy number aberrations associated with invasion and recurrence.

Citation Format: Kaile Wang, Tapsi Kumar, Junke wang, Darlan Minussi, Emi Sei, Jianzhuo li, Tuan Tran, Aatish Thennavan, Min Hu, Anna Casasent, Zhenna Xiao, Shanshan Bai, Yuehui Zhao, Amado Zurita, Ana Aparicio, Brian Chapin, Jie ye, Jianjun Zhang, Don Gibbons, Andrew Futreal, Lorraine King, Jeffrey Marks, E. Shelley Hwang, Vandna Shah, Ellinor Sawyer, Petra Kristel, Jelle Wesseling, Esther H. Lips, Nicholas Navin. Archival single cell sequencing reveals persistent subclones over years to decades of DCIS 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 125.
Abstract 1253: Dietary tryptophan catabolite released by intratumoralLactobacillus reuterifacilitates aPD-L1 therapy
Authors: Bender Mackenzie , McPherson Alex , Phelps Catherine , Davar Diwakar , Vignali Dario A. , Zarour Hassane , Meisel Marlies
Published: April 2023

The use of probiotics by cancer patients is increasing, including amongst those undergoing immune checkpoint inhibitor (ICI) therapy. Here, we elucidate a critical microbial-host crosstalk between probiotic-released aryl hydrocarbon receptor (AhR) agonist indole-3-aldehyde (I3A) and CD8 T cells within the tumor microenvironment that potently enhances spontaneous antitumor immunity and facilitates ICI therapy in preclinical melanoma. Our study reveals that probiotic Lactobacillus reuteri (L.reuteri) translocates to, colonizes, and persists within melanoma, where via its released dietary tryptophan catabolite I3A, it locally promotes interferon-γ-producing CD8 T cells, thereby facilitating ICI therapy efficacy. Moreover, L.reuteri secreted I3A was both necessary and sufficient for antitumor immunity, and loss of AhR signaling within CD8 T cells abrogated L.reuteri’s ability to restrain melanoma growth. Further, a tryptophan-enriched diet potentiated both L.reuteri- and ICI-induced antitumor immunity. Finally, we provide evidence for a potential role of I3A in promoting ICI therapy efficacy and survival in advanced melanoma patients.

Citation Format: Mackenzie Bender, Alex McPherson, Catherine Phelps, Diwakar Davar, Dario A. Vignali, Hassane Zarour, Marlies Meisel. Dietary tryptophan catabolite released by intratumoralLactobacillus reuterifacilitates aPD-L1 therapy [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 1253.
Abstract 13: A non-surgical approach to develop an orthotopic colorectal cancer mouse model
Authors: Sharma Samantha , Van der Jeught Kevin , Zhou Zhuolong , Zhang Xinna , Lu Xiongbin
Published: April 2023

Recapitulating physiologically relevant tumor microenvironment is an important yet a significant challenge in modeling colorectal cancer (CRC) for basic and translational studies. Cecal-wall injection is most widely used for orthotopic animal models of CRC. However, this procedure involves complicated surgical procedures, and its ectotopic nature is a major issue for therapeutic development. Here, we report a non-surgical procedure for establishing orthotopic CRC mouse models, reflecting the appropriate tumor microenvironment to better model the disease development and facilitate translational studies, including drug identification and validation. For this procedure, the mice are fasted overnight. The following day, anesthesia is induced in the study mice (Balb/c or C57BL/6) while placed on a thermostat-controlled heating blanket. Next, a specialized catheter is inserted and distended into the mouse colon to gently block the colon and ensure the localization for tumor development. Hereafter, the colonic epithelium is treated with proteases and mechanical abrasion to cause mild disruption. Following the local gut inflammation, the targeted colon region is incubated with tumor cells, leading to the adhesion of tumor cells (CT26 or MC38) to the colonic epithelium. Based on the animal background and the cell line used, the tumor is developed within 4-6 weeks. Pathological assessment on hematoxylin and eosin stained tumor tissues confirmed the stages of tumor development. Additional adaptations of this procedure include i) tumor development in different mouse strains, ii) local delivery of adeno-Cre in transgenic mice for CRC, and iii) transplantation of tumor organoids in the mouse colon. Overall, our non-surgical procedure overcomes the technical difficulties of surgery and offers several advantages like ease of handling, cost-effectiveness, pain-free with nearly no procedure-associated mortality, and adaptability to several strains. To conclude, this non-surgical procedure is a novel and promising alternative to conventional orthotopic CRC mouse models for translational studies.

Citation Format: Samantha Sharma, Kevin Van der Jeught, Zhuolong Zhou, Xinna Zhang, Xiongbin Lu. A non-surgical approach to develop an orthotopic colorectal cancer mouse model [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 13.
Abstract 1317: Exercise training prevents exosome mediated pre-metastatic niche formation in the liver
Authors: Yang Ruiqi , Haykal Tony , Tohme Celine , Wang Ronghua , Kaltenmeier Christof , Geller David , Yazdani Hamza , Tohme Samer
Published: April 2023

Introduction: Primary tumor cells release exosomes facilitating pre-metastatic niche formation in distant organs, initiating the process of metastasis. As metastasis is the main prognostic factor for patients, it is essential to understand the pre-metastatic niches and develop therapies to target them. It has been shown that Kupffer cells play an crucial role in the formation of pre-metastatic niche in the liver. We have recently shown that exercise training (ExT) alters the phenotype of Kupffer cells and attenuates tumor metastasis after surgery. Here, we hypothesize that ExT can interfere with exosome mediated pre-metastatic niches, hence decreasing metastasis.

Methods: Eight-week-old male mice were randomized into sedentary and ExT group. Mice were pre-educated with cancer exosomes for 4 weeks to induce pre-metastatic niche and intra-splenically injected with murine colorectal cancer (MC38) cells. Tumor metastasis and growth was followed by 3 weeks.

Results: Compared to the sedentary group, there was a significant decrease in metastatic nodules and hepatic tumor replacement in the ExT mice liver. To further study the effects of ExT, livers from both groups were harvested 4 weeks after exosome pre-education. We observed decrease in the uptake of tumor exosome, immune cells infiltration and fibronectin deposition in the ExT livers, which demonstrates less pre-metastatic niche formation. Bulk RNA sequencing was performed on Kupffer cells isolated from both groups. Ingenuity pathway analysis shows that Triggering Receptor Expressed on Myeloid cells 1 (TREM1) signaling pathway was downregulated in EXT group, indicating less activation of Kupffer cells.

Conclusions: These results indicate that exercise reduces pre-metastatic niche formation and tumor metastasis by affecting the interaction between cancer exosomes and Kupffer cells. In light of these findings, potential may exist to reduce exosome-induced liver pre-metastatic niches through the use of ExT as a nonpharmacologic therapy in patients with cancer.

Citation Format: Ruiqi Yang, Tony Haykal, Celine Tohme, Ronghua Wang, Christof Kaltenmeier, David Geller, Hamza Yazdani, Samer Tohme. Exercise training prevents exosome mediated pre-metastatic niche formation in the liver [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 1317.
Abstract 1344: myCAFs and iCAFs have similar lineage
Authors: Walter Thomas S. , Han Lu , Ostrowski Michael
Published: April 2023

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a dense stroma primarily composed of cancer associated fibroblasts (CAFs). These fibroblasts are a diverse population of cells; some groups of fibroblasts have been shown to play tumor supporting roles while others have been shown to be tumor suppressing. Two prolific and well-characterized CAF subtypes—myofibroblasts (myCAFs) and inflammatory CAFs (iCAFs)—could potentially have different regulatory effects on tumor growth. Understanding the tissue origin of these two groups of CAFs could assist in understanding the complex and heterogeneous roles they might play in PDAC development. Previous studies in our group showed that the splanchnic mesenchyme, a layer of mesenchyme adjacent to the fetal pancreatic epithelium, gives rise to the majority of the total CAF population. Here, I aim to test the hypothesis that both iCAFs and myCAFs originate from the splanchnic mesenchyme. myCAFs have been previously observed to be proximal to tumor cells and express αSMA at a higher level, while iCAFs have been observed to be distal to tumor cells and express αSMA at a lower level. Here, we used a genetically engineered mouse PDAC model carrying KrasG12D/+;p53Frt/+;Pdx1FlpO/+;Isl1cre/+;R26Tomato/+ alleles. In this model, epithelial specific FlpO expression leads to Kras activation and p53 deletion, causing tumorigenesis. Simultaneously, splanchnic specific Cre expression leads to permanent tomato expression in the splanchnic descendants. The pancreata of these mice were harvested, fixed, and mounted onto slides. Co-immunostaining with αSMA and tomato was then done. The stained slides were scanned, and the Inform software was used to segment and phenotype the cells. More than 95% of αSMA high cells were tomato positive, and more than 95% of αSMA low cells were also tomato positive. We thus conclude that both myCAFs and iCAFs originate from the splanchnic mesenchyme. Consistent with previous in vitro studies, this lineage tracing study suggests that different CAF subtypes are likely due to patterning influenced by different factors within the tumor microenvironment.

Citation Format: Thomas S. Walter, Lu Han, Michael Ostrowski. myCAFs and iCAFs have similar lineage [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 1344.