bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1999: The impact of the COVID-19 pandemic on 30-day survival in pediatric cancer patients in low, middle, and high-income countries: an international, observational cohort study
Authors: Elhadi Muhammed , Bandyopadhyay Soham , Peter Noel , Lakhoo Kokila ,   Global Children's NCDs Collaborative
Published: June 2022

Objectives: To conduct the first international cohort study to ascertain the short-term outcome for pediatric oncology patients who underwent treatment across 16 high-income countries (HICs) and 23 low- and- middle-income countries (LMICs) during the COVID-19 pandemic. The hypotheses being tested was that the COVID-19 pandemic had affected paediatric cancer care, and that the outcomes of children were worse in LMICs.

Design: A multicenter, international, collaborative cohort study.

Setting: 91 hospitals and cancer centers in 39 countries providing cancer treatment to pediatric patients between March and December 2020.

Participants: Patients were included if they were under the age of 18 years, and newly diagnosed with or undergoing active cancer treatment for Acute lymphoblastic leukemia, Non-Hodgkin lymphoma, Hodgkin lymphoma, Wilms Tumor, Sarcoma, Retinoblastoma, Gliomas, Medulloblastomas or Neuroblastomas, in keeping with the World Health Organization Global Initiative for Childhood Cancer.

Main outcome measure: All-cause mortality at 30 days and 90 days.

Results: 1660 patients were recruited. Over 30 days, 45 LMIC patients (4.3%; 95% CI: 3.1 to 5.5) and 2 HIC patients (0.4%; 95% CI: -0.1 to 0.9) died. 219 children had their treatments delayed, interrupted, or modified. LMIC patients had 11.7 (95% CI: 10.3 to 13.1) and 7.4 (95% CI: 6.5 to 8.3) times the risk of death at 30 days and 90 days respectively (p < 0.001). After adjusting for confounders, pediatric cancer patients in LMICs had 35.7 times the odds of death at 30 days (p < 0.001).

Conclusions: The COVID-19 pandemic has affected pediatric oncology service provision. It has disproportionately affected patients in LMICs, highlighting and compounding existing disparities in healthcare systems globally that need addressing urgently. However, most pediatric cancer patients continued to receive their normal standard of care. This speaks to the adaptability and resilience of health-care systems and healthcare workers globally.

Citation Format: Muhammed Elhadi, Soham Bandyopadhyay, Noel Peter, Kokila Lakhoo, Global Children's NCDs Collaborative. The impact of the COVID-19 pandemic on 30-day survival in pediatric cancer patients in low, middle, and high-income countries: an international, observational cohort study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1999.
Abstract 219: Towards targeted-gold nanoparticle enhanced chemoradiation therapy of pancreatic cancer
Authors: Papineni Rao V. , Adhikary Amitava , Bhattacharya Santanu
Published: June 2022

The goal of this project is to develop a unique pancreatic cancer treatment platform where, X-ray excited Targeted-Gold nanoparticles (T-GNPs) trigger and enhance drug action (I-RaGAZ, Integrated radiation combined with T-GNP and 2-Azido-2-deoxy-D-glucose (2-AZ-2-DG)). Herein, we plan to utilize a novel three stage nanotechnology-chemoradiation combinatorial approach for enhancing catastrophic cell death in pancreatic tumors and reducing normal tissue toxicity. T-GNP has cytotoxic effect at tumor site and can specifically sensitize tumor tissue for radiation. In this work, we used our GNP-based pancreatic ductal adenocarcinoma (PDAC) -specific drug delivery platform using a plectin-1-targeting peptide (PTP) (patent: US2018/0333432 A1) to demonstrate the cytotoxicity of the nanocomplex in vitro. In PANC-1 cancer cell model, using traditional gold-standard clonogenic assay, we show 2-AZ-2-DG has radiation-mediated cytotoxicity which is further augmented by T-GNP. These results align with our hypothesis and the electron spin resonance (ESR) spectroscopy electron-induced aminyl radical production data presented here. This radiation-produced auger electrons from T-GNP likely increased formation of RNH• which result in enhanced cell kill. This via H-atom abstraction reactions that might cause DNA-strand breaks and/or crosslink formation leading to lesions that could induce apoptosis of cancer cells.

This nanotool paraphrased as I-RaGAZ, Integrated radiation induced T-GNP - 2-AZ-2-DG anti-tumor action, will be very useful for treating deep-seated tumors with minimum collateral damage. This approach, integrated radiation-mediated azido tumor cure (I-RaGAZ), has vast global cancer treatment potential considering the dual mode of damaging the tumor tissues and protecting the normal tissues/organs.

Citation Format: Rao V. Papineni, Amitava Adhikary, Santanu Bhattacharya. Towards targeted-gold nanoparticle enhanced chemoradiation therapy of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 219.
Abstract 2194: Rapid interrogation of colon cancer driver genes in vivo using split Cre and CRISPR-Cas9
Authors: Zhang Daniel , Westcott Peter , Naranjo Santiago , Hauck Haley , Rideout William , Sacks Nate , Bhutkar Arjun , Patten Justin , Goto Norihiro , Sedlak Joseph , Jacks Tyler
Published: June 2022

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths worldwide, with a five-year survival rate of only 63% with current treatment regimens. While genetically engineered mouse models (GEMMs) have contributed widely to our understanding of the underlying genetics and molecular mechanisms of the disease, major limitations exist. Most models of CRC lack flexibility to initiate focal adenocarcinomas with different genetic perturbations within the native microenvironment of the distal colon. Specifically, no autochthonous model of metastatic CRC compatible with in vivo CRISPR-Cas9 genome engineering has been described. Therefore, we developed a novel split Cre-recombinase system restricting activation of oncogenic KrasG12D and deletion of the tumor suppressor gene (TSG) Trp53 to Lgr5+ stem cells of the colon crypts. Importantly, these events are not only critical drivers of CRC progression and metastasis in mice and humans but can also lead to non-specific transformation of the surrounding colonic stroma. By expressing one half of Cre from the endogenous Lgr5 locus and other half via colonoscopy-guided submucosal injection of lentivirus, we restricted Cre-mediated recombination to enterocytes. Lentivirus was also engineered to express a single guide RNA (sgRNA) against Apc, facilitating tumor initiation via deletion of this critical CRC TSG in mice expressing Cas9. A second sgRNA was included to facilitate rapid interrogation of putative cancer driver genes. As proof-of-principle, we targeted Smad4, a critical suppressor of metastasis in CRC, and observed adenocarcinoma formation and liver metastases with complete recombination and editing of all targeted genes. To prioritize candidate TSGs for future in vivo interrogation, we applied MutSigCV to large human sequencing datasets and identified genes significantly co-mutated with APC and KRAS in CRC. Altogether, this faithful and tractable model of CRC can be used for rapid functional interrogation of TSGs and should accelerate downstream mechanistic and preclinical studies.

Citation Format: Daniel Zhang, Peter Westcott, Santiago Naranjo, Haley Hauck, William Rideout, Nate Sacks, Arjun Bhutkar, Justin Patten, Norihiro Goto, Joseph Sedlak, Tyler Jacks. Rapid interrogation of colon cancer driver genes in vivo using split Cre and CRISPR-Cas9 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2194.
Abstract 2231: Surface-enhanced Raman scattering paltform of nanometal for exosome analysis in hepato-cellular carcinoma
Authors: Ahmed Ola Sayed , imam hisham , zekri abdel Rahman
Published: June 2022

Exosomes could be a source of cancer biomarkers in the future. Exosomes derived from tumors could provide a non-invasive way to diagnose cancer, assess cancer progression, and track treatment responses. However, exosome analysis would be aided by new molecular methods, which would speed up basic and clinical exosome research. Exosome research is rapidly advancing, providing new insights into the physiological role of exosomes and their significant link to human health. Although tumor-derived exosomes have been shown to be promising biomarkers for cancer detection and disease progression due to their inherited biological contents from the parent cancer cells and unique roles in tumor metastasis and invasion, rapid and effective isolation from complex biological samples and high-precision real-time analysis remains a difficult task. In this work, we present a surface-enhanced Raman scattering (SERS) platform for integrating multiple hepatocellular carcinoma exosomes conventional method to read different peak positions and amplitudes of spectrum, whole SERS spectra of exosomes were analyzed by principal component analysis (PCA). In addition, label-free analysis of purified exosomes from normal cell and hepatocellular carcinoma were clearly distinguished from normal cell derived exosomes by 96.4% sensitivity and 98.2% specificity using surface-enhanced Raman spectroscopy (SERS). In this study, the possibility of using exosomes as biomarkers for the detection of hepatocellular carcinoma (HCC) was discussed. From this study, SERS can serve as a valuable tool for separating exogenous subpopulations and can be used to characterize exosomes for cancer diagnosis.

Citation Format: Ola Sayed Ahmed, hisham imam, abdel Rahman zekri. Surface-enhanced Raman scattering paltform of nanometal for exosome analysis in hepato-cellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2231.
Abstract 2239: Detecting cancer when it can be cured: The potential for cure across all stageable cancers
Authors: Hubbell Earl , Clarke Christina
Published: June 2022

Introduction: Early detection of cancer may reduce cancer mortality by providing access to treatments with the potential to cure cancer at early stages. A mixture cure model divides cancer cases into two populations: one where cancer is likely to severely impact mortality (not-cured) and one where long-term survival with low risk is possible (cured). Previous work on such models has concentrated on estimating cure for either many cancer types without regard to stage (public health arena), or single cancers by stage at diagnosis (screening arena). A gap in the current literature is an estimation of likelihood of cure in many cancer types at all stages.

Methods: We estimate a mixture-cure model for all stages simultaneously for stageable cancers using the fact that cancer is a progressive disease (worse by stage) as a constraint on model parameters. We applied this estimation procedure to 21 cancer classes with standard American Joint Committee on Cancer (AJCC) staging using 12 years of cancer-specific survival data in 50-79 year old individuals using Surveillance, Epidemiology and End Results (SEER) program data from 2006-2015 followed to 2018. For each stage, we recover the fraction of those cured (“cure fraction”, i.e., long-term survivors with minimal excess hazard) and not-cured (severe acute mortality due to cancer, modeled as a Weibull distribution).

Results: Cure fractions varied between cancer classes. Two important types of cancer behavior are illustrated by colorectal and gallbladder cancers. Colorectal cancer had a good potential for cure at any stage before metastasis, with a precipitous drop from 63% (95% CI: 62-64%) cure at stage III to 7% (6-7%) cure at stage IV. In contrast, gallbladder cancer exhibited a systematic decrease at each stage, with 47% (43-52%) cure fraction at stage I, 22% (20-24%) at stage II, 20% (17-22%) at stage III, and 2% (2-3%) at stage IV. Differences in 5-year survival between earlier stages and metastasis were highly correlated to differences in cure fraction (r^2=0.97), suggesting differences in 5-year survival are a proxy for differences in cure.

Conclusions: Long-term survivors were evident at early stages for all 21 cancer types examined. These survival fractions were all greatly reduced by the time cancer reached metastasis. This indicates early-stage cancers do not differ from late-stage cancers simply by lead time and provides statistical evidence that detection of cancer in early stages may result in long-term survival for many stageable cancer types.

Citation Format: Earl Hubbell, Christina Clarke. Detecting cancer when it can be cured: The potential for cure across all stageable cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2239.
Abstract 2316: Malignant potential of colorectal adenoma based on the telomere length
Authors: Tomasova Kristyna , Kroupa Michal , Siskova Anna , Horak Josef , Vycital Ondrej , Liska Vaclav , Vodickova Ludmila , Vodicka Pavel
Published: June 2022

Telomere attrition is a well-known hallmark of malignancy in cancer tissue. Our study analyzed the relative telomere length in adenomas, precursor lesions of colorectal adenocarcinoma. We investigated if the accelerated telomere length shortening occurs before potential malignant transformation to adenocarcinoma and corresponds with clinicopathological characteristics.

Relative telomere length was measured in 145 patients using multiplex qPCR. We have recorded significant shortening of telomeres in the adenoma tissue compared to adjacent mucosa (Wilcoxon Signed Rank test, p < 0.0001). Further, we have observed a weak inverse relationship between both adenoma and mucosa telomere length and age (Spearman's and Pearson's correlation, p = 0.021 and p = 0.027, respectively), contrary to our previous study of colorectal cancer patients (n = 696), where the inverse correlation of telomere length was detected only in the non-malignant mucosa (Pearson's correlation, p = 0.029). However, the telomere length ratio of adenoma to adjacent mucosa did not correspond with the degree of dysplasia according to the Vienna classification (Kruskal-Wallis test, p = 0.572). Also, there was no difference in the telomere length ratio among histological types of adenomas and no association with the adenoma localization in proximal and distal colon or rectum (Kruskal-Wallis test, p = 0.648 and p = 0.459, respectively).

Preliminary results indicate that telomere shortening occurs indeed as early as in a stage of adenomatous polyps, and it is less pronounced in adenomas compared to cancerous cells. However, the relative telomere length does not seem to correspond with adenoma progression towards malignancy. In planned experiments, we intend to assess mitochondrial DNA damage caused by 8-oxoguanine DNA adducts. These major oxidative base lesions can arise from endogenous reactive oxygen species and impact, among other things, the integrity of telomeres through double-strand breaks to DNA. Mitochondrial DNA is more prone to accumulation of oxidative damage than nuclear DNA and thus might be useful as an early, additional indicator of the precancerous character of adenoma. The study will reveal whether increased mitochondrial DNA damage and telomere loss are commonly present and linked together in adenomas.

This study was supported by the Charles University (project GA UK No. 120, UNCE/MED/006) and the Grant Agency of the Czech Republic (21-04607X, 21-27902S).

Citation Format: Kristyna Tomasova, Michal Kroupa, Anna Siskova, Josef Horak, Ondrej Vycital, Vaclav Liska, Ludmila Vodickova, Pavel Vodicka. Malignant potential of colorectal adenoma based on the telomere length [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2316.
Abstract 2416: Ras-mediated activation of mTORC2 drives breast cancer cell migration and invasion
Authors: Wiegand Mollie E. , Collins Shannon E. , Brown Isabella N. , Werner Alyssa N. , Mundo Isabelle M. , Mouneimne Ghassan , Charest Pascale G.
Published: June 2022

We previously identified the mechanistic Target of Rapamycin Complex 2 (mTORC2) as an effector of Ras for the control of directed cell migration in Dictyostelium. Recently, this relationship was found to be conserved in mammalian cells, and mTORC2 was shown to be an effector of oncogenic Ras. Deregulated mTORC2 signaling in cancer is mostly associated with increased survival and metabolic reprogramming, but mTORC2 has also been linked to cancer cell migration, particularly in breast cancer. Interestingly, although Ras is rarely mutated in breast cancer, it is often upregulated due to amplification and overexpression of growth factor receptors. Here, we investigated the role of Ras in promoting the migration and invasion of breast cancer cells through mTORC2. We observed that Ras and mTORC2 promote the migration of breast cancer cells, independently of the breast cancer molecular subtype. Using breast epithelial MCF10A cells transformed with HER2 or mutant Ras, we found that Ras promotes mTORC2 activation, and mTORC2-dependent migration and invasion. We further observed that while mutant Ras-transformed MCF10A cells display uncontrolled cell proliferation and an invasive phenotype, silencing of the mTORC2-dependent component Rictor mostly leads to loss of invasiveness. Together, our findings suggest that, whereas Ras activation of mTORC2 is likely to play a minor role in breast tumor formation, the Ras-mTORC2 pathway plays a key and general role in promoting the migration and invasion of breast cancer cells.

Citation Format: Mollie E. Wiegand, Shannon E. Collins, Isabella N. Brown, Alyssa N. Werner, Isabelle M. Mundo, Ghassan Mouneimne, Pascale G. Charest. Ras-mediated activation of mTORC2 drives breast cancer cell migration and invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2416.
Abstract 2418: Markers for metastasis evaluation of the ErbB family and GDF15 inratosteosarcoma
Authors: Ishiyama Shun , West Bailey , Han Hyo-Jeong , Guo Xin , Ito Tomoaki , Saeki Harumi , Brock Malcolm V. , Gabrielson Kathleen
Published: June 2022

The most recent advance in the treatment of osteosarcoma (OS) occurred in the 1980s when multi-agent chemotherapy was shown to improve overall survival compared to surgery alone. Animal model investigations can accelerate the understanding of human diseases and since few models are available for OS, we refined a rat model to ask questions regarding pathways of metastasis and metastasis biomarker predictiveness. The outbred Sprague-Dawley (SD) rat model is an immunocompetent, syngeneic model, as we implant UMR106 OS cells (ATCC), which were originally isolated from a SD OS. We developed a reproducible method to orthotopically implant cells into the tibia microenvironment and to amputate the limb for longitudinal studies. Using young rats, we also modeled pediatric OS. We found that rats develop reproducible primary and metastatic pulmonary tumors which histologically resemble human primary and metastatic osteosarcoma. Using immunohistochemistry methods, we found that primary OS is infiltrated with numerous macrophages with relatively fewer CD3+ T cells. A protein expression survey of these OS cells revealed significant expression of ErbB family receptor tyrosine kinases (ErbB2, ErbB4), which have recently been shown to be anti-inflammatory against macrophages. We also discovered that rats with osteosarcoma have markedly high serum levels of Growth Differentiation Factor 15 (GDF15), also called Macrophage Inhibitory Cytokine 1, a member of the TGF beta family of proteins, which is also frequently expressed with ErbB2 in cancers. GDF15 was measured by ELISA. Western blot analysis showed that GDF15 precursor (pro-peptide) was expressed in primary and metastases. This model will enable studies with anti-ErbB2 or GDF15 pathway inhibition to assess the role of macrophage activation in the setting of pathway inhibition and chemotherapy, as well as how pathway inhibition modulates metastasis incidence. In vivo and invitro, systems will enable the assessment of the role of GDF15 as a potential biomarker of metastasis, compared to tibial radiographic patterns, serum alkaline phosphatase, and lung histopathology. In conclusion, we have an immunocompetent, syngeneic orthotopic osteosarcoma rat model which affectively resembles human osteosarcoma, and can be used for better understanding metastasis mechanisms and metastasis biomarkers.

Citation Format: Shun Ishiyama, Bailey West, Hyo-Jeong Han, Xin Guo, Tomoaki Ito, Harumi Saeki, Malcolm V. Brock, Kathleen Gabrielson. Markers for metastasis evaluation of the ErbB family and GDF15 inratosteosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2418.
Abstract 2420: Exploring associations between the IL4 pathway and colorectal cancer risk in in vivo models and clinical records
Authors: Beckstead Jacob , Petros Jack , Mehrotra Kunaal , Fingleton Barbara
Published: June 2022

We have previously demonstrated that inhibition of the IL4 pathway limits metastatic potential in colorectal cancers and have postulated that the type II IL4 receptor could serve as a potential target in risk assessment and treatment. Here we expand on these findings through in vivo mouse studies and retrospective patient analysis. To elucidate how colorectal cancers may use the IL4 pathway to grow and infiltrate surrounding tissue, we performed two in vivo studies independently targeting IL4ra in myeloid and intestinal epithelial tissues. Mice with a deleted allele of IL4ra and a floxed allele of IL4ra were crossed with either a line expressing a myeloid-specific Cre (LysM Cre) or an intestinal epithelial-specific Cre (Villin Cre). Our results indicate that myeloid-expressed IL4ra facilitates tumorigenesis but is not a significant contributor to later growth. In contrast, epithelial IL4ra significantly contributes to tumor number and size, but had little effect on the observed number of aberrant crypt foci, an early indicator of colon cancer.

We hypothesized that effects of atypical IL4 activity could also be observed in the medical record. Atopic diseases such as asthma and eczema are characterized by an overabundance of IgE, a protein regulated by the IL4 pathway. Using Vanderbilt University Medical Center’s SD Discover program, we obtained de-identified records for patients diagnosed with at least stage II colorectal carcinoma between 2005 and 2015. We then stratified patients by metastatic progression. Patients who were diagnosed with asthma prior to age sixty were significantly less likely to develop an advanced form of colorectal cancer and at reduced risk of metastatic disease or recurrence. This may be a result of medication prescribed for atopic disease such as inhalers and corticosteroids and we are performing further analyses to test these hypotheses.

Citation Format: Jacob Beckstead, Jack Petros, Kunaal Mehrotra, Barbara Fingleton. Exploring associations between the IL4 pathway and colorectal cancer risk in in vivo models and clinical records [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2420.
Abstract 2423: A Cdh3 - Lam332 signaling axis by a unique subset of leader cells controls breast tumor organoid protrusion dynamics and directed collective migration
Authors: Cao Yanyang , Hwang Priscilla Y. , Clarke Maria , Almeida Jose , Pathak Amit , Longmore Gregory D.
Published: June 2022

Carcinoma dissemination can occur when heterogeneous tumor and tumor stromal cells clusters migrate together via collective migration. Cells at the front lead and direct collective migration, yet how these leader cells form and interact with the microenvironment to direct migration are not fully appreciated. From live videos of primary mouse and human breast tumor organoids in a 3D microfluidic system that mimics the native breast tumor microenvironment, we developed 3D computational models. These hypothesize that for leader cells to polarize to the leading edge and lead directed collective migration they need to generate high protrusive forces and overcome extracellular matrix (ECM) resistance. From scRNAseq of migrating primary tumor organoids, we identify significant K14+ “leader” cell heterogeneity that differs depending upon the environmental signal. We isolate a unique Cadherin-3 (Cdh3) positive leader cell subpopulation that is necessary and sufficient to lead collective migration. Cdh3 controls leader cell protrusion dynamics through controlling the local production of Laminin-332 which is required for integrin/focal adhesion function. Loss of Cdh3 expression in mouse and human primary breast tumor organoids and invasive breast tumor cell lines significantly impairs directed collective migration. In the absence of leader cell Cdh-Cdh3 intercellular adhesions there is reduced cellular β-catenin levels and Laminin 332 production. In syngeneic, orthotopic transplant mouse models, Cdh3 depletion in tumor cells resulted in decreased laminin 332 production at the invasive edge of primary tumors and decreased lung metastasis. Chromatin Immunoprecipitation experiments revealed that in contacted cells β-catenin and TCF4 are present on Lam α3, β3, and γ2 promoter regions. Laminin 332 production by leader cells was required for optimal Integrin/FA function and cellular protrusion stability. These results indicated that in leader cells local Cdh3-Cdh3/β-catenin regulated Laminin 332 production controls protrusion dynamics and overcomes resistance to ECM to lead directed collective tumor migration. Our findings highlight how a unique subset of leader cells in breast tumors interact with the microenvironment to direct collective migration.

Citation Format: Yanyang Cao, Priscilla Y. Hwang, Maria Clarke, Jose Almeida, Amit Pathak, Gregory D. Longmore. A Cdh3 - Lam332 signaling axis by a unique subset of leader cells controls breast tumor organoid protrusion dynamics and directed collective migration [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2423.
Abstract 2431: A novel role of MUC5AC/CD44v6/c-Met axis in breast cancer brain metastasis
Authors: Maurya Shailendra Kumar , Siddiqui Jawed A. , Gautama Shailendra K. , Kanchan Ranjana K. , Pothuraju Ramesh , Natarajan Gopalakrishnan , Atri Pranita , Venkata Ramakanth C. , Bhatia Rakesh , Khan Parvez , Rehmana Asad Ur , Chaudhary Sanjib , Perumal Naveenkumar , Mahapatra Sidharth , Chand Hitendra S. , Jain Maneesh , Santamaria-Barriab Juan A. , Cittelly Diana M. , Batra Surinder K. , Nasser Mohd W.
Published: June 2022

Brain metastasis (BrM) is one of the leading causes of mortality in breast cancer (BC). Although BrM is associated with all BC subtypes, but it is more prevalent in triple-negative and HER2+ BC patients. Recent advancements in the multimodality treatment of primary BC have prolonged patient survival, which has in turn increased the incidence of BrM. Due to the lack of understanding of BC-BrM, there is no reliable biomarker and effective therapeutic strategy. In this regard, using publicly available databases and RNA-Seq analysis, we observed that secretory mucin MUC5AC is significantly upregulated in BC brain metastatic cell lines and tissues compared to their respective control. We validated these observations in brain-seeking BC (BSBC) cell lines and brain metastatic tissues at protein levels. Interestingly, we found that the MUC5AC levels were significantly increased in the serum of CNS metastatic patients relative to healthy donors and BC patients. Our functional studies revealed that silencing of MUC5AC in BSBC cell lines showed a significant decrease in cell adhesion, migration, brain metastatic potential, and increased survival of mice. Exploring the molecular mechanism further revealed that MUC5AC interacts with CD44v6 and c-Met and deletion of MUC5AC demonstrated a decrease in the expression of c-MET and CD44v6. Furthermore, pharmacological targeting of MUC5AC through c-Met inhibitor reduces the expression of CD44v6 and MUC5AC, suggesting that c-Met inhibitors could be used as a novel therapeutics for targeting MUC5AC and thereby attenuating BC brain metastasis. Altogether, our study demonstrates that MUC5AC/CD44v6/c-Met axis could be used as a novel approach to prevent breast cancer brain metastasis.

Citation Format: Shailendra Kumar Maurya, Jawed A. Siddiqui, Shailendra K. Gautama, Ranjana K. Kanchan, Ramesh Pothuraju, Gopalakrishnan Natarajan, Pranita Atri, Ramakanth C. Venkata, Rakesh Bhatia, Parvez Khan, Asad Ur Rehmana, Sanjib Chaudhary, Naveenkumar Perumal, Sidharth Mahapatra, Hitendra S. Chand, Maneesh Jain, Juan A. Santamaria-Barriab, Diana M. Cittelly, Surinder K. Batra, Mohd W. Nasser. A novel role of MUC5AC/CD44v6/c-Met axis in breast cancer brain metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2431.
Abstract 2460: Assessing the effectiveness of transarterial chemoembolization using super-resolution ultrasound imaging and a rat model of hepatocellular carcinoma
Authors: Li Junjie , Brown Katherine , Yociss Megan , Eisenbrey John , Hoyt Kenneth
Published: June 2022

Introduction: Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related deaths worldwide. Transarterial chemoembolization (TACE) treatment of HCC restricts the blood supply to the tumor. Conventional assessment of tumor response is performed by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) or computed tomography (DCE-CT) at 4 to 6 weeks after the TACE procedure. An earlier indication of treatment effectiveness would improve patient management and prognosis. Super-resolution ultrasound (SR-US) imaging allows a ten-fold improvement in spatial resolution compared to traditional ultrasound (US) and allows visualization of microvascular networks. The goal of this preclinical research study was to use SR-US imaging to assess the effectiveness of TACE treatment in a relatively brief time frame.

Methods: Eight male Sprague Dawley rats weighing around 300 g were injected with 5 million N1S1 cells in the upper left liver lobe and the tumors were allowed to grow for 14 days. The TACE procedure consisted of transhepatic arterial delivery of a mixture of 25 µL Lipiodol and 25 µL Doxorubicin (10 mg/kg) via a polyethylene microcatheter. After intravenous injection of a microbubble (MB) contrast agent, contrast-enhanced US images were acquired (N = 3000) with a preclinical system (Vevo 3100, FUJIFILM VisualSonics Inc) equipped with an MX201 linear array transducer. US images were processed to remove high motion frames, followed by tissue suppression filtering, and then MB localization and accumulation. Morphological filtering was used to enhance the vessel structures. In vivo US, MRI, and CT imaging were performed at baseline before TACE and again at 1 and 2 wk. After euthanasia, tumor tissue was removed for ex vivo analysis.

Results: A rat model of HCC for assessing TACE treatment was introduced. Based on tumor size changes and residual perfusion after the TACE procedure, treatments were determined to be a complete responder, partial responder, or non-responder. Initial results demonstrate that SR-US imaging can sensitively detect any reduction in HCC perfusion as measured by a microvascular density (MVD) metric within 2 wk after a single TACE procedure. MVD measurements from the SR-US images were consistence with tumor volume data from MRI and CT imaging.

Conclusions: In vivo SR-US images of tumor microvascular networks provided insight into treatment efficacy.

Citation Format: Junjie Li, Katherine Brown, Megan Yociss, John Eisenbrey, Kenneth Hoyt. Assessing the effectiveness of transarterial chemoembolization using super-resolution ultrasound imaging and a rat model of hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2460.
Abstract 252: STAT3 signaling in the tumor microenvironment promotes primary gastric cancer growth and metastasis formation
Authors: Eissmann Moritz , Huber Anne , Allam Amr , Dijkstra Christine , Ernst Matthias
Published: June 2022

Tumor cell intrinsic activation of Signal Transducer and Activator of Transcription 3 (STAT3) signaling has been shown to promote stemness, tumorigenesis and cancer therapy resistance. Therefore, Stat3 inhibitors are being developed and tested pre-clinically and in clinical trials for multiple solid cancers. Importantly, STAT3 signaling also occurs in the non-tumor cells within the tumor microenvironment (TME). The effect of Stat3 signaling within the TME on primary gastric cancers (GC), metastasis formation and therapy responses is unknown.

Here, we describe a novel genetically defined, tumor organoid-driven, transplantable, orthotopic gastric cancer model that represents gastric carcinomas with associated metastasis formation in immune competent recipient mice. Utilizing our murine GC organoid model in subcutaneous allograft experiments, we showed that established GC tumors respond to pharmacological Stat3 inhibition. This anti-tumor effect is primarily driven by inhibition of the Stat3 signaling in the TME as tumor growth was increased in Stat3 high TME recipients, while tumor growth was inhibited in Stat3-signaling reduced Stat3+/- mice. Furthermore, Crisp-Cas9 -mediated Stat3 knockout in the GC organoids did not alter their in vitro growth potential. Stat3KO GC allograft tumor growth was not changed compared to Stat3WT GC tumors grown in wildtype host mice. Collectively, this suggests that inhibition of Stat3 signaling within the TME but not the tumor cell intrinsic Stat3 reduction confer anti-tumor responses. Through gastric serosa transplantation of the murine GC organoids into Stat3-elevated gp130FF mutant mice, we confirmed that high Stat3 signaling within the TME promotes distal metastasis formation to the liver.

Taken together, we show that Stat3 signaling represents a therapy target, that predominantly acts through the TME. Better understanding of the signaling within the TME in primary and metastatic GC lesions will allow identification of novel tumor cell and TME-centric therapy targets. Combination of therapies targeting tumor cell intrinsic vulnerabilities with novel pro-tumorigenic TME -targeting agents could help to improve responses against primary and metastatic disease and therefore improve outcome for the worst prognosis patients.

Citation Format: Moritz Eissmann, Anne Huber, Amr Allam, Christine Dijkstra, Matthias Ernst. STAT3 signaling in the tumor microenvironment promotes primary gastric cancer growth and metastasis formation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 252.
Abstract 2531: a
Authors: Rajesh Adarsh , Havas Aaron , Varanasi Siva Karthik , Evans Katrina , Cheikh Bassem B. , Duerrschmid Clemens , Mammadova Najiba , Adams Peter , Kaech Susan
Published: June 2022

Hepatocellular carcinoma (HCC) is an emerging health crisis due to rapidly increasing incidence rates and an overall 5-year survival rate of 18%. Consequently, there is an urgent need for new therapies and better preventive strategies. Aging and obesity are two of the biggest risk factors for HCC, and with ever-increasing aging and obese populations, uncovering how aging and obesity drive the increased risk for HCC has become imperative. Aging and obesity are well known to be characterized by immune dysregulation leading to persistent inflammation, and a dysregulated immune microenvironment is also a common theme shared by multiple hallmarks of HCC. In this study, we aim to assess the immune microenvironment of aged and obese mouse livers by performing spatial phenotyping of distinct immune signatures which can potentially predispose hepatocytes to HCC. To address this, we used Akoya’s ultra-high multiplexed CODEX (CO-Detection by indEXing) imaging system to interrogate 31 different biomarkers in liver samples at the single cell level. Specifically, we compared the liver immune microenvironment of mice within 2 different treatment groups (old mice on a high fat diet (HFD) and young mice on a normal diet (ND)) to the immune landscape of liver tumor tissue. Our goal is to identify distinct immune phenotypes present in both, liver tumor tissues and livers of HFD mice, compared to ND livers - to better understand how immune dysregulation can predispose an aged and obese liver to HCC.

Citation Format: Adarsh Rajesh, Aaron Havas, Siva Karthik Varanasi, Katrina Evans, Bassem B. Cheikh, Clemens Duerrschmid, Najiba Mammadova, Peter Adams, Susan Kaech. a [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2531.
Abstract 2573: PI3K/AKT/mTOR pathway and CDK4/6 inhibitors efficiently inhibit cell growth of HPV-positive and HPV-negative head and neck squamous cell carcinoma in vitro
Authors: Demers Imke , Verhees Femke , Leegemaate Dion , Jacobs Robin , Hoeben Ann , Hoebers Frank , Kremer Bernd , Speel Ernst-Jan
Published: June 2022

Introduction: Both HPV-positive and -negative head and neck squamous cell carcinoma (HNSCC) often show activation of the PI3K/AKT/mTOR pathway, due to, amongst others, PI3KCA mutations, PTEN loss, or receptor tyrosine kinase activation. In HPV-negative tumors, CDKN2A (p16) inactivation or CCND1 (cyclin D1) amplification frequently occurs resulting in sustained cyclin dependent kinase (CDK) 4/6 activation. The aim of our study was to investigate the efficacy of PI3K/AKT/mTOR pathway inhibitors (PI3Ki) (alpelisib, buparlisib and gedatolisib) and CDK4/6 inhibitors (CDKi) (palbociclib and ribociclib) in HPV-positive and -negative HNSCC cell lines.

Methods: The efficacy of the inhibitors was assessed using MTT assays and changes in PI3K and Cyclin D1/CDK pathway protein expression were determined by Western blotting. Cell cycle analysis was performed with flow cytometry and apoptosis was assessed by Annexin-V staining. Changes in cell metabolism were assessed by Seahorse XF assays.

Results: Both HPV-positive and -negative HNSCC cell lines were highly sensitive to the PI3Ki (Gedatolisib, IC50 5-30 nM > Buparlisib, IC50 0.6-3.6 µM > Alpelisib, IC50 3-23 µM). In general, PI3Ki decreased pathway activity, resulted in moderate cell cycle arrest and apoptosis, and decreased oxidative and glycolytic metabolism. CDKi were particularly effective in blocking HPV-negative cell line growth (Palbociclib, IC50 0.5-2 µM > Ribociclib, IC50 4-7 µM). CDK inhibition showed decreased pRb expression and G1 cell cycle arrest, whereas apoptosis was not induced.

Conclusion: PI3Ki and CDKi efficiently inhibit their respective pathways and HNSCC cell growth in vitro, the latter only in HPV-negative cell lines. Whereas PI3Ki especially show an effect on oxidative and glycolytic metabolism, CDKi particularly lead to cell cycle arrest. Further research should elucidate whether these inhibitors may be effective therapeutic agents in HNSCC patients.

Citation Format: Imke Demers, Femke Verhees, Dion Leegemaate, Robin Jacobs, Ann Hoeben, Frank Hoebers, Bernd Kremer, Ernst-Jan Speel. PI3K/AKT/mTOR pathway and CDK4/6 inhibitors efficiently inhibit cell growth of HPV-positive and HPV-negative head and neck squamous cell carcinoma in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2573.
Abstract 2670: ERAS-601, a potent allosteric inhibitor of SHP2, synergistically enhances the efficacy of sotorasib/adagrasib and cetuximab in NSCLC, CRC, and HNSCC tumor models
Authors: Martin Leenus , Patel Roopal , Zhang Jingchuan , Nevarez Robin , Congdon Taylor , Brail Les , Shoemaker Robert
Published: June 2022

ERAS-601 is a potent, small molecule allosteric inhibitor of wildtype SHP2, a non-receptor protein tyrosine phosphatase (PTP) encoded by the PTPN11 gene. ERAS-601 inhibits wild type SHP2 biochemically with an IC50 of 4.6 nM and demonstrates selectivity across panels of 300 kinases and 12 phosphatases. SHP2 mediates upstream receptor tyrosine kinase (RTK) signaling via its phosphatase-mediated regulation of guanine nucleotide exchange factors (GEFs). ERAS-601 inhibits the SHP2 dependent cycling of KRAS from the inactive GDP-bound state to the active GTP-bound state and demonstrates anti-proliferative activity in KRASG12C and EGFR amplified cell lines. The combination of upstream blockade of RAS-GTP cycling by ERAS-601 with inhibition of KRASG12C by a selective KRASG12C inhibitor synergistically inhibits cellular proliferation in multiple KRASG12C mutated human cancer cell lines. The combinations of ERAS-601 with KRASG12C inhibitors achieve tumor growth inhibition that is superior to the respective ERAS-601 and KRASG12C monotherapies in NSCLC and CRC CDX and PDX tumor models. Similarly, the combination of ERAS-601 with an EGFR antibody, cetuximab, inhibits oncogenic RAS/MAPK signaling as measured by pERK1/2 and enhances the anti-proliferative activity of cetuximab in RAS/RAF wild type CRC and HPV-negative HNSCC cell lines. The combination of ERAS-601 with cetuximab achieves tumor growth inhibition that is superior to respective ERAS-601 and cetuximab monotherapies in RAS/RAF wild type CRC and HPV-negative HNSCC CDX and PDX tumor models. These nonclinical data support the clinical development of ERAS-601 in combination with a KRASG12C inhibitor in NSCLC and CRC tumors, and ERAS-601 in combination with cetuximab in RAS/RAF wild type CRC as well as HPV-negative HNSCC tumors. Both combinations are being studied in ongoing clinical studies (HERKULES-2, NCT04959981; FLAGSHP-1, NCT04670679).

Citation Format: Leenus Martin, Roopal Patel, Jingchuan Zhang, Robin Nevarez, Taylor Congdon, Les Brail, Robert Shoemaker. ERAS-601, a potent allosteric inhibitor of SHP2, synergistically enhances the efficacy of sotorasib/adagrasib and cetuximab in NSCLC, CRC, and HNSCC tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2670.
Abstract 2859: Platelet-derived TGFβ undermines treatment efficacy of t cell recruiting bispecific antibodies
Authors: Lutz Martina S. , Klimovich Boris , Maurer Stefanie , Heitmann Jonas S. , Maerklin Melanie , Zekri Latifa , Jung Gundram , Salih Helmut R. , Hinterleitner Clemens
Published: June 2022

Immunotherapeutic strategies such as T cell-recruiting bispecific antibodies (bsAb), chimeric antigen receptor (CAR) T cells, and immune checkpoint inhibition have revolutionized oncological treatment. However, many patients do not respond to treatment with the so far available therapeutics, others for limited time only. Various tumor immune evasion mechanisms have been reported to counteract efficiency of T cell engaging therapeutics. Thrombocytes have recently been described to affect cancer biology by mediating epithelial-mesenchymal transition, tumor invasion, metastasis and notably also immune evasion. Here we report that, upon treatment of prostate cancer patients within a clinical trial evaluating a novel PSMAxCD3 bsAb (NCT04104607), we observed profound treatment-associated platelet activation, mirrored by a decrease of total platelet count. Upon modelling the treatment setting, we found that platelet activation significantly reduced bsAb-mediated CD4+ and CD8+ T cell reactivity as revealed by impaired T cell activation, reduced secretion of perforin and ultimately inhibition of target cell lysis. The immunosuppressive effect mediated by platelets occurred in a TGFβ-dependent manner and was not restricted to the PSMAxCD3 bsAb, but was also observed with various other CD3-directed bispecific constructs including the clinically approved CD19xCD13 bsAb Blinatumomab. BsAb-mediated T cell reactivity could generally be restored by inhibiting platelets using dabigatran, but also specifically by blocking the TGFβ axis. Together, our findings unravel that platelets undermine the efficacy of T cell-recruiting bsAb and identify modulation of platelet function and TGFβ as means to reinforce the effectiveness of bsAb treatment.

Citation Format: Martina S. Lutz, Boris Klimovich, Stefanie Maurer, Jonas S. Heitmann, Melanie Maerklin, Latifa Zekri, Gundram Jung, Helmut R. Salih, Clemens Hinterleitner. Platelet-derived TGFβ undermines treatment efficacy of t cell recruiting bispecific antibodies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2859.