bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of Mast Cell Degranulation in Atopic Dermatitis by Celastrol through Suppressing MRGPRX2
Authors: Yao Ciyu , Ye Wenzhen , Chen Mengxue
Published: January 2023

Background. Atopic dermatitis is a common dermatological disease, and mast cell degranulation is believed to be related with the progression of atopic dermatitis. Mas-related G protein-coupled receptor-X2 (MRGPRX2), and calcium release-activated calcium channel protein 1-2 (ORAI-1, ORAI-2) are involved in mast cell degranulation. Celastrol is an active monomer of Tripterygium wilfordii, and it presents an antiatopic role. Methods. 2,4-Dinitrofluorobenzene (DNFB) and compound 48/80 (C 48/80) were used to establish a slow and acute scratching animal model, respectively. Hematoxylin-eosin and toluidine blue staining was used to investigate tissue injury. Inflammatory factor concentration was measured with ELISA. The expression of MRGPRX2, ORAI-1, and ORAI-2 was detected with immunohistochemistry (IHC) staining. Gene expression profiling and microRNA array were performed to investigate gene differential expression. Results. Celastrol greatly inhibited atopic dermatitis-related tissues injury, mast cell production, histamine release, scratching level, inflammatory factor expression, and activation of MRGPRX2/ORAI axis in the DNFB-induced atopic dermatitis model. The influence of Celastrol on atopic dermatitis was remarkably reversed by overexpression of MRGPRX2. Conclusion. We found that the improvements of atopic dermatitis caused by Celastrol were reversed by treatment with MRGPRX2OE, indicating that Celastrol might affect atopic dermatitis through MRGPRX2. This study might provide a novel thought for the prevention and treatment of atopic dermatitis by regulating MRGPRX2.
Laboratory Characteristics on SARS-CoV-2 Infection among Patients with Diabetes Mellitus: A Single-Center Retrospective Study
Authors: Salem Raneem O. , Nuzhat Ayesha , Zaheer Shawana , Kallash Majd Aldeen
Published: January 2023

Background. Diabetic patients have been severely affected by COVID-19 infection. It has been reported that the disease is more progressive leading to venous and arterial thromboembolism, due to multiple factors. This study was conducted to determine the hematologic parameters including D-dimer in diabetic patients with COVID-19 infection in association with disease severity and treatment. Method. This retrospective cohort study was conducted at King Fahad Medical City, Saudi Arabia, after obtaining IRB approval, by collecting data regarding all laboratory parameters, disease severity, and anticoagulant treatment of COVID-19 diabetic patients (

n
=
159

) from medical records from March to December 2020. Result. Mean value of white blood cells, neutrophils, monocytes, eosinophils, lymphocyte monocyte ratio, C-reactive protein, serum ferritin, and LDH levels was elevated in severe cases than in mild cases with statistical significant increase in HbA1c (0.047), serum fibrinogen (0.007), C-reactive protein (0.005), serum ferritin (0.034), and serum LDH (0.015). Mortality was observed in 14 (8.8%) patients mostly with severe COVID-19 with diabetes. In our study, treatment with low molecular weight heparin was not significantly related to severity. A logistic regression analysis indicated an association of some laboratory parameters with severity and mortality of the disease. Conclusion. The routine blood parameters if detected early will enable physicians to identify severe cases of COVID-19 patients with Diabetes for prompt treatment and save considerable time and resources.
Dual mTORC1/2 Inhibition Synergistically Enhances AML Cell Death in Combination with the BCL2 Antagonist Venetoclax
Authors: Satta Toshihisa , Li Lin , Chalasani Sri Lakshmi , Hu Xiaoyan , Nkwocha Jewel , Sharma Kanika , Kmieciak Maciej , Rahmani Mohamed , Zhou Liang , Grant Steven
Published: January 2023

Purpose:

Acute myelogenous leukemia (AML) is an aggressive disease with a poor outcome. We investigated mechanisms by which the anti-AML activity of ABT-199 (venetoclax) could be potentiated by dual mTORC1/TORC2 inhibition.

Experimental Design:

Venetoclax/INK128 synergism was assessed in various AML cell lines and primary patient AML samples in vitro. AML cells overexpressing MCL-1, constitutively active AKT, BAK, and/or BAX knockout, and acquired venetoclax resistance were investigated to define mechanisms underlying interactions. The antileukemic efficacy of this regimen was also examined in xenograft and patient-derived xenograft (PDX) models.

Results:

Combination treatment with venetoclax and INK128 (but not the mTORC1 inhibitor rapamycin) dramatically enhanced cell death in AML cell lines. Synergism was associated with p-AKT and p-4EBP1 downregulation and dependent upon MCL-1 downregulation and BAK/BAX upregulation as MCL-1 overexpression and BAX/BAK knockout abrogated cell death. Constitutive AKT activation opposed synergism between venetoclax and PI3K or AKT inhibitors, but not INK128. Combination treatment also synergistically induced cell death in venetoclax-resistant AML cells. Similar events occurred in primary patient-derived leukemia samples but not normal CD34+ cells. Finally, venetoclax and INK128 co-treatment displayed increased antileukemia effects in in vivo xenograft and PDX models.

Conclusions:

The venetoclax/INK128 regimen exerts significant antileukemic activity in various preclinical models through mechanisms involving MCL-1 downregulation and BAK/BAX activation, and offers potential advantages over PI3K or AKT inhibitors in cells with constitutive AKT activation. This regimen is active against primary and venetoclax-resistant AML cells, and in in vivo AML models. Further investigation of this strategy appears warranted.
Abstract A018: Abiraterone vs. enzalutamide: Does age impact one treatment over another
Authors: Nikita Nikita , Shaver Amy , Gandhi Krupa , Keith Scott W. , Sharma Swapnil , Lu-Yao Grace
Published: January 2023

Background: Abiraterone and Enzalutamide are two of the most commonly used treatments for prostate cancer (PCa). Currently, both these medications are approved for similar clinical indications even though they have different toxicity profiles. This study aims to identify if patients and providers prefer one medication to another and if the choice depends on age. Methods: We used SEER-Medicare data to identify primary PCa patients diagnosed between 1999 and 2017. We excluded those diagnosed at death. Abiraterone and enzalutamide use was identified using part D files. Patients had 6 months continuous part D coverage prior to treatment. Age was parameterized categorically: 65-74 years, 75-84 years, and over 85 years. Descriptive statistics were generated and evaluated by Chi-square tests for all categorical variables. Multivariable logistic regression was used to investigate disparities between the treatments adjusting for demographic characteristics. Results: We identified 8,398 patients with PCa over 65 years of age; 4,180 used abiraterone and 4,218 used enzalutamide. The demographic characteristics for both groups were similar (p = 0.1557). Most patients were 65-74 years old (62.6% in the abiraterone group and 62.4% in the enzalutamide group), followed by approximately 30% of each group in the 75-84 age range. Finally, the enzalutamide group had 8.2% 85 years and older, while the abiraterone group had 7.1%. The appeared to be no preference in odds of use of either medication by age group. When comparing the very old (85+) to the 65-74 year olds (OR=0.906; 95% CI 0.745, 1.101; p=0.3727). Nor was there a statistical significance when comparing odds of use between 75-84-year-olds to those who were 65-74 (OR=0.978; 95%CI 0.873, 1.095; p=0.6927). Conclusion: Our study found that abiraterone and enzalutamide use among patients does not differ substantially by age groups. Further studies are warranted to understand these associations better.

Citation Format: Nikita Nikita, Amy Shaver, Krupa Gandhi, Scott W. Keith, Swapnil Sharma, Grace Lu-Yao. Abiraterone vs. enzalutamide: Does age impact one treatment over another [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr A018.
Abstract A021: Older adult-specific microbes correlate with treatment response and markers of T-cell senescence in NSCLC
Authors: Spakowicz Daniel , Hoyd Rebecca , Wheeler Caroline E. , Williams Nyelia , Bibi Amna , Husain Marium , Rajamouli Srichandhana , Suman Shankar , Amann Joseph , Grogan Madison , Vibhakar Pooja , Owen Dwight H. , Carbone David P. , Rosko Ashley , Burd Christin E. , Presley Carolyn J.
Published: January 2023

This abstract is being presented as a short talk in the scientific program. A full abstract is available in the Short Talks from Proffered Abstracts section (PR004) of the Conference Proceedings.

Citation Format: Daniel Spakowicz, Rebecca Hoyd, Caroline E. Wheeler, Nyelia Williams, Amna Bibi, Marium Husain, Srichandhana Rajamouli, Shankar Suman, Joseph Amann, Madison Grogan, Pooja Vibhakar, Dwight H. Owen, David P. Carbone, Ashley Rosko, Christin E. Burd, Carolyn J. Presley. Older adult-specific microbes correlate with treatment response and markers of T-cell senescence in NSCLC [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr A021.
Abstract B001: Elevated all-cause and cardiovascular mortality in cancer survivors with sarcopenia
Authors: Braithwaite Dejana , Karanth Shama , Leeuwenburgh Christiaan , Manini Todd M. , Wheeler Meghann , Yang Danting , Aduse-Poke Livingstone , Anton Stephen , Penedo Frank , Siegel Erin M. , Licht Jonathan D. , Zhang Dongyu
Published: January 2023

Abstract: Sarcopenia, a condition characterized by the loss of muscle mass, strength and function with age, is highly prevalent in cancer survivors. The relationship between sarcopenia and prognosis among cancer survivors is not well understood. Methods: From the Third National Health and Nutrition Examination Survey (NHANES III), we identified 946 participants who were diagnosed with cancer (mean age 60.6 years); the most common disease sites were breast, prostate, colon and melanoma. We assembled an age, sex and race-matched cohort of 1,857 participants without cancer (mean age, 60.2 years). Sarcopenia was defined by appendicular lean mass and body height (men <7.26 kg/m2, women <5.45 kg/m2). Proportional-hazard models were used to assess whether sarcopenia was associated with all cause and CVD-specific time to death after taking into account baseline age, sex, race/ethnicity, smoking status, and energy intake for each cohort. In the cancer survivor cohort, models were additionally adjusted for a history of having more than one cancer and time since diagnosis. Mortality was ascertained from the National Center for Health Statistics Linked Mortality Files. Results: There were 321 deaths among cancer survivors (33.9% of the cohort) during a median follow-up of 10.5 years versus 495 deaths among participants without cancer (26.7% of the cohort) during a median follow-up of 10.9 years. Deaths from cardiovascular disease (CVD) were observed in 58 (6.1%) cancer survivors versus 122 (6.6%) participants without cancer. Overall, sarcopenia was more prevalent among cancer survivors versus the matched cohort (22.2% versus 19.7% respectively). Rates of CVD death were more than twice higher among cancer survivors with versus without sarcopenia (adjusted hazard ratio, 2.17, 95% confidence interval [CI], 1.16 to 4.05). All-cause mortality was 79% higher (adjusted hazard ratio, 1.79; 95% CI, 1.36 to 2.36) among cancer survivors with versus without sarcopenia. No significant associations were seen between sarcopenia and rates of death from all causes and CVD specifically among participants without cancer. Conclusion: Cancer survivors with sarcopenia have an increased risk of all-cause and CVD-specific mortality. In contrast, sarcopenia was not a major predictor of these outcomes in the matched cohort without cancer.

Citation Format: Dejana Braithwaite, Shama Karanth, Christiaan Leeuwenburgh, Todd M. Manini, Meghann Wheeler, Danting Yang, Livingstone Aduse-Poke, Stephen Anton, Frank Penedo, Erin M. Siegel, Jonathan D. Licht, Dongyu Zhang. Elevated all-cause and cardiovascular mortality in cancer survivors with sarcopenia [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr B001.
Abstract IA011: Mutations in normal cell
Authors: Stratton Mike
Published: January 2023

Mutations in DNA/RNA cause the phenotypic variation upon which natural selection acts during evolution. Mutations arise and are transmitted between generations of organisms and between generations of cells in the somatic tissues of multicellular organisms. However, our knowledge of the biological processes that generate mutations and the rates at which they occur in different cell types has been rudimentary until recently. Cancer is a recognized outcome of somatic mutations. However, we have limited understanding of the wider consequences of somatic mutations. I will review our current understanding of the landscape of somatic mutations in normal cells, its causes and its consequences.

Citation Format: Mike Stratton. Mutations in normal cell [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr IA011.
Abstract IA013: Age-related changes in pancreatic fibroblasts drive pancreatic cancer growth and progression
Authors: Zabransky Daniel , Jaffee Elizabeth , Weeraratna Ashani
Published: January 2023

Aging is an independent risk factor for the development of pancreatic ductal adenocarcinoma (PDAC). However, the ways in which aging alters cell populations that will ultimately comprise the PDAC tumor microenvironment (TME) are incompletely understood. PDAC tumors are highly desmoplastic, and fibroblasts alter the behavior of pancreatic cancer cells and impact treatment responses. Here, we show that normal pancreatic fibroblasts acquire tumor-promoting properties during aging using both in vitro and in vivo models. To determine if PDAC tumors may be influenced by an aged microenvironment, we performed orthotopic KPC cell injections into the pancreata of aged (>52-week-old) and young (6-8-week-old) syngeneic C57Bl/6J mice. Aged mice have significantly larger tumors, increased incidence of liver metastases, and decreased survival compared to young mice. We next queried whether aged pancreatic fibroblasts contribute to this increase in tumor growth. We generated a panel of aged (donors >55 years old) and young (donors <35 years old) normal human pancreatic fibroblasts to determine the effects of healthy aging fibroblasts on PDAC cell behavior. Conditioned media from aged human pancreatic fibroblasts significantly increases the proliferation of PDAC cells, suggesting that secreted factors produced by aged fibroblasts enhance cancer cell growth. Next, we evaluated changes in the invasiveness of PDAC cells in a 3D co-culture system with either aged or young pancreatic fibroblasts. PDAC spheroids containing cancer cells and aged fibroblasts exhibit a significant increase in invasiveness compared to those containing young fibroblasts. Aged (>52-week-old) compared to young (6-8-week-old) mouse pancreatic fibroblasts also increase the proliferation and invasiveness of KPC cells in vitro, providing evidence that aged fibroblasts may contribute to the phenotypes observed in vivo experiments using aged mouse models. In conclusion, we show that aged, non-cancer-associated pancreatic fibroblasts have the potential to promote growth, migration, and invasiveness in multiple models of pancreatic ductal adenocarcinoma. We hypothesize that factors secreted by pancreatic fibroblast change during aging and that this aged secretome may serve to establish a tumor-promoting niche for pancreatic cancer cells.

Citation Format: Daniel Zabransky, Elizabeth Jaffee, Ashani Weeraratna. Age-related changes in pancreatic fibroblasts drive pancreatic cancer growth and progression [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr IA013.
Abstract IA020: Increased mitochondrial DNA mutation burden with age limits lung cancer
Authors: White Eileen
Published: January 2023

Mitochondrial DNA (mtDNA) mutations accumulate with age and are found in some cancers. However, the relationship between cancer and mtDNA mutations, and the pathogenicity of those mutations, is not known. To address this question, we created a new mouse model of non-small cell lung cancer (NSCLC) with an increased mtDNA mutation burden. This model was generated by breeding the NSCLC mice model carrying floxed alleles of p53 and KrasG12D, with the mtDNA mutator mouse bearing a germline polymerase gamma mutation that disables proofreading function (POLGD257A) that is responsible for the accumulation of mtDNA mutations over time. The mutator mice with NSCLC had reduced tumor burden and prolonged survival after tumor induction in comparison to POLG wild type mice with NSCLC. Both tumor and normal tissue from mutator mice had elevated mtDNA mutation burden in comparison to wild type mice. The prolonged survival of POLGD257A mice with NSCLC was even more pronounced in older animals where mtDNA mutation burden was further increased. The same was true for a mouse model with a single pathogenic mtDNA mutation found in mitochondrial disease, indicating that it is not the number of mutations, but rather the disabling of mitochondrial function by mutation that causes reduced tumor growth. This suggested that elevated mtDNA mutation with age may suppress NSCLC. To address the mechanism by which accumulation of mtDNA mutations suppress tumor growth we examined metabolism and immune functions. We found that mtDNA mutations caused tumor cell metabolic dysfunction including increased glycolysis and decreased respiration with dependency of glycose and serine and glycine as carbon sources in vitro. Moreover, mtDNA mutations caused tumor dependence on serine and glycine in vivo. Thus, elevated mtDNA mutation burden with age may increase probability of suppression of mitochondrial function, resulting in specific metabolic dependencies in NSCLC. Thus, tumors with mtDNA mutation accumulation may be susceptible to therapeutic dietary restriction of serine and glycine.

Citation Format: Eileen White. Increased mitochondrial DNA mutation burden with age limits lung cancer [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr IA020.
Abstract IA023: Age against the machine: How the aging microenvironment drives tumor progression
Authors: Weeraratna Ashani
Published: January 2023

Patients 65 and older account for 69% of all new cancer diagnoses. Melanoma is a deadly disease, with chances of survival for patients with stage IV disease totaling only 27%. Age-related changes- both secreted and biophysical- drive metastasis, immune cell recruitment, and changes in the vasculature. Age-related changes also impact the ability of tumor cells to respond to targeted therapy, and new data from our lab indicate this may be sex specific. Our recent studies also show that age-related changes in the lung microenvironment facilitate a permissive niche for efficient outgrowth of disseminated dormant tumor cells, in contrast to the aged skin, where age-related changes suppress melanoma growth but drive dissemination. We now show that age-induced changes in distal metastatic microenvironments promote efficient reactivation of dormant melanoma cells in the lung due to changes in Wnt signaling pathways. Overall, our studies argue for placing mechanistic studies of cancer biology into the context of aging.

Citation Format: Ashani Weeraratna. Age against the machine: How the aging microenvironment drives tumor progression [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr IA023.
Abstract PR002: Cytokines surge in the blood years prior to a cancer diagnosis in elderly individuals
Authors: Chen Guangbo , Zheng Hong , Rosenberg-Hasson Yael , Araya Saron , Padilla Cindy , Sarin Kavita , Dekker Cornelia L. , Grant Philip , Maecker Holden T. , Furman David , Shen-Orr Shai , Khatri Purvesh , Davis Mark M.
Published: January 2023

How the human immune system reacts prior to a cancer diagnosis is largely unknown. Here we analyze a longitudinal cohort of 133 healthy individuals (18 to over 90 years old), 28 of whom developed cancers over nine years. We find the global abundance of a broad spectrum of cytokines (26 out of 32 assayed) surges 2-3 years before cancer diagnosis but not with other conditions (such as inflammatory or cardiovascular diseases). These cytokine surges precede cancer diagnosis only in those 80 years or older, suggesting age-specific inflammatory effects. Meanwhile, in early-stage (but not late-stage) tissues across 10 cancer types, aging elevates cytokine transcriptional activity, which coincides with an age-dependent expansion of the inflammatory macrophage population. We further find that macrophage is a major producer of cytokines in cancer tissues, irrespective of cancer types. These results suggest that cancer associated macrophage is a major source of peripheral inflammatory molecules and may mobilize immunity well before a clinical diagnosis in the elderly.

Citation Format: Guangbo Chen, Hong Zheng, Yael Rosenberg-Hasson, Saron Araya, Cindy Padilla, Kavita Sarin, Cornelia L. Dekker, Philip Grant, Holden T. Maecker, David Furman, Shai Shen-Orr, Purvesh Khatri, Mark M. Davis. Cytokines surge in the blood years prior to a cancer diagnosis in elderly individuals [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr PR002.
Abstract A004: The role of calcium in metastatic progression
Authors: Morris Kenise , Papa Anne-Laure
Published: January 2023

Calcium is essential to the structural stability of integrin proteins implicated in platelet–cancer cell interactions, namely integrins GPIIb/IIIa and avb3, which share a common beta subunit (Pelletier et al., J Biol Chem, 1996), (Zhang and Chen, Cell Adh Migr, 2012). Additionally, calcium is essential for ligand binding to these integrins (e.g., fibrinogen) that bridge platelets and cancer cells (Raborn et al., Biochemistry, 2011) and for the downstream signaling following mechanical and chemical stimulation in the integrin microenvironment (Michelson, Platelets, 2013). Platelet-cancer cell interactions are key players in cancer progression as platelets support metastatic dissemination (Morris et al., Biochim. Biophys. Acta BBA - Rev. Cancer, 2022). Despite previous studies recognizing calcium’s role in integrin stability and function, calcium’s importance for platelet support in cancer progression is not well understood. Therefore, we aim to (1) characterize the receptor expression and ligand binding ability of GPIIb/IIIa and avb3 on respective cells via anti-CD41a (for platelets and breast cancer MDA-MB-231 cells) and anti-avb3 antibody (for lung cancer A549 cells), (2) characterize platelet function by assessing platelet aggregation using light transmission aggregometry and platelet adhesion on a thrombogenic surface, (3) characterize platelet-cancer cell interaction via flow cytometry, and (4) assess cancer cell invasion in the presence of platelets via transwell assay; all in the presence of an environment with calcium chelation and hypercalcemic conditions. Our results demonstrate that platelet GPIIb/IIIa, cancer cell GPIIb/IIIa (MDA-MB-231) and cancer cell integrin avb3 (A549) are modulated with calcium chelation. Additionally, platelet function, specifically aggregation is significantly decreased with calcium chelation and significantly increased with hypercalcemia. We also observed that the interaction between platelets and cancer cells is reduced with calcium chelation, platelet-MDA-MB-231 interaction specifically being significantly reduced in the presence of sodium citrate. Our overall hypothesis is that the observed effect of calcium levels of platelet and cancer cell integrins could have a significant influence on platelet-cancer cell interactions and cancer cell migration in hypercalcemia, often seen in patients with malignancy. A greater understanding of these interactions could have impact for the design of specific inhibitors with anti-metastatic effects and potentially lead to novel targeted therapies to combat metastatic dissemination.

Citation Format: Kenise Morris, Anne-Laure Papa. The role of calcium in metastatic progression [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A004.
Abstract A005: Adherent-to-suspension transition by hematopoietic factors promotes metastatic dissemination
Authors: Huh Hyunbin D. , Gee Heon Yung , Park Hyun Woo
Published: January 2023

How primary tumor cells reprogram their anchorage dependency to disseminate into the bloodstream remain elusive. Here, we discover a biological phenomenon referred to as “Adherent-to-Suspension Transition” (AST) that reprograms adherent cells into suspension cells by introducing four hematopoietic transcriptional regulators, which we designated AST factors. Induction of AST factors in adherent cells suppress Hippo transducers YAP-TEAD and upregulate globin genes that evoke spontaneous cell-matrix dissociation and anoikis resistance in the absence of lineage differentiation. During dissemination, we uncover the aberrant induction of AST factors in circulating tumor cells (CTCs) derived from de novo metastatic breast cancer patients and mouse models in vivo. Pharmacological blockade of AST factors in breast cancer cells abrogates CTC formation and suppresses lung metastases without affecting the primary tumor growth. Collectively, we demonstrate that suspension cells can directly arise from adherent cells by the addition of defined factors that confer metastatic traits. Furthermore, our data expand the prevailing cancer treatment paradigm toward direct intervention within the metastatic spread of cancer.

Citation Format: Hyunbin D. Huh, Heon Yung Gee, Hyun Woo Park. Adherent-to-suspension transition by hematopoietic factors promotes metastatic dissemination [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A005.
Abstract A010: MYC as a master regulator of dormancy in triple negative breast cancer
Authors: Roelofs Charlotte , Mouchemore Kellie A. , Chakrabarti Anannya , Redvers Richard P. , Anderson Robin L.
Published: January 2023

Dormancy is a major clinical problem in breast cancer. Although the five-year survival rate is high, 15% of the patients will relapse. Cancer cells that disseminate from the tumor (DTCs) can enter a dormant state that allows survival in the face of standard-of-care therapies. These DTCs are the source of cancer recurrence. Currently little is known about the molecular drivers that initiate dormancy. MYC is a well-known oncogene that is aberrantly regulated in many cancers, including breast cancer. It is involved in proliferation, tumorigenesis, and diapause. We propose that MYC is also a master regulator of dormancy. To investigate its role in dormancy, we generated cell lines with regulatable levels of MYC expression. Using an in vitro dormancy assay, we have shown that reduced MYC levels induce dormancy in otherwise aggressive cell lines. In mice bearing human breast tumors, the loss of MYC activity following primary tumor removal greatly reduces the extent of metastasis. The DTCs in lungs, livers, spine, and femurs are maintained as small clusters with few cells, while control mice have many and large metastases. MYC reduction maintains the DTCs in this dormant-like state for at least 32 days after primary tumor removal. Importantly, when MYC levels are restored, the DTCs exit the dormant state, and overt metastasis ensues. Currently, we are comparing the transcriptomes of dormant DTCs to actively growing metastatic cells to generate a MYC-driven dormancy signature that will elucidate potential dormancy genes to target therapeutically in the clinic. To complement our genetic studies, we are investigating pharmacological suppression of MYC activity. The bromodomain inhibitor I-BET151 reduces MYC protein levels and induces dormancy in our in vitro assay. Mice treated with I-BET151 after primary tumor removal had significantly reduced levels of metastasis in lung, liver, and spine when compared to their non-treated counterparts. Testing of this compound in an immune-competent preclinical model is currently underway. Our data imply that MYC is an important regulator of dormancy in breast cancer. Our future experiments will elucidate dormancy genes as potential targets that could lead to life-saving therapies in the clinic.

Citation Format: Charlotte Roelofs, Kellie A. Mouchemore, Anannya Chakrabarti, Richard P. Redvers, Robin L. Anderson. MYC as a master regulator of dormancy in triple negative breast cancer [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A010.
Abstract A012: NAT10 promotes metastasis through enhancer remodeling of cancer cells
Authors: Amin Ruhul , Ha Ngoc-Han , Meier Jordan L. , Hunter Kent W.
Published: January 2023

Distant organ metastases account for the majority of the cancer-related death. However, the detailed mechanism underlying the metastatic potential of cancer cells is poorly understood. We have previously shown than NUP210-dependent nuclear pore-associated chromatin-bound protein complex is involved in cellular mechanotransduction and metastasis in breast cancer. Although NAT10, an RNA cytidine acetyl transferase has been shown to interact with this protein complex, specific function of NAT10 in metastasis is not well understood. Using allograft and genetically engineered highly metastatic PyMT mouse model, we found that the loss of NAT10 significantly decreases lung metastasis in mice. NAT10 is mainly localized to the nucleolus and nucleoplasmic compartment of the cancer cells. Current understanding suggests a role of NAT10 in acetylation of mRNAs and ribosomal RNAs. In contrast to its role as an RNA acetylase, we uncovered a new mechanism of NAT10 in cancer cells. Using co-immunoprecipitation followed by mass spectrometry and chromatin immunoprecipitation followed by high throughput sequencing (ChIP-seq), we found that NAT10 is interacting with super-enhancer associated protein complex. RNA-seq analysis revealed that the loss of NAT10 leads to dramatic transcriptional suppression of super-enhancer driven genes including MYC. Mechanistically, using mass spectrometry-based quantification of protein acetylation, we showed that the loss of NAT10 leads to decreased acetylation of enhancer-associated Histone H4 and H3 modification. NAT10 loss also leads to increased heterochromatin-forming histone methylation within the cell nuclei. Our study revealed a distinct role of NAT10 as an enhancer remodeler in metastatic cancer cells. Therefore, NAT10 could be a potential therapeutic target in preventing metastatic disease.

Citation Format: Ruhul Amin, Ngoc-Han Ha, Jordan L. Meier, Kent W. Hunter. NAT10 promotes metastasis through enhancer remodeling of cancer cells [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A012.
Abstract A022: Role of imaging mass cytometry in studying hepatoblastoma vascular invasion
Authors: Espinoza Andres F. , Patel Roma , Woodfield Sarah , Vasudevan Sanjeev A.
Published: January 2023

Introduction: Hepatoblastoma (HB) is the most common liver cancer in children with a survival rate of less than 50% in patients with high-risk disease, especially those with vascular invasion (VI). Patients with VI receive aggressive therapies that are toxic and ineffective; therefore, a better understanding of the biomarkers and biology of VI in HB is urgently needed. Methods: Using imaging mass cytometry (IMC) we analyzed 18 pre-validated protein targets on two of our formalin-fixed paraffin embedded (FFPE) HB patient samples; one with VI and one without VI. For confirmation of the proteins of interest, we performed immunohistochemistry (IHC) on primary and metastatic lesions from a separate patient. We performed RNA sequencing of one of our primary liver tumor and two sub-clones of that tumor, one inside an intrahepatic vessel and one sample representing tumor vessel thrombus. Results: By IMC, we found that protein expression of c-Myc and YAP-1 are increased in the areas of VI as the tumor invades the neighboring blood vessel. YAP-1 and c-Myc expression were low in the patient without VI. When examining the IHC of the separate patient with metastatic lung disease, we noted clear nuclear positivity of YAP-1 and c-Myc in the metastatic cells, compared to the negative nature of the primary liver tumor. Our RNA sequencing data showed that the average gene expression of c-Myc of the tumor in intrahepatic vessel and tumor vessel thrombus was 1.61 times higher than the primary tumor. The tumor found in the intrahepatic vessel and tumor vessel was found to have 1.29 times higher expression of Yap-1 compared to the primary tumor. Conclusions: YAP-1 and c-Myc had higher expression in hepatoblastoma as the tumor cells progress to invade the vasculature and metastasize. YAP-1 and c-Myc expression appears to play a role in vascular invasion that is maintained when tumors grow in the metastatic sites.

Citation Format: Andres F. Espinoza, Roma Patel, Sarah Woodfield, Sanjeev A. Vasudevan. Role of imaging mass cytometry in studying hepatoblastoma vascular invasion [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A022.
Abstract A043: A novel role for Mitochondrial Rho GTPase 2 in tumor cell invasion and metastasis
Authors: Boulton Dillon P. , Furnish Madison , Caino M. Cecilia
Published: January 2023

Metastasis of cancer cells to secondary sites remains the leading cause of cancer related deaths, emphasizing a strong need for actionable targets in advanced stage cancer. To address this, we study novel dysregulated signaling mechanisms that cells utilize to metastasize to distal sites. Here, we focus on how outer mitochondrial membrane protein—Mitochondrial Rho GTPase 2 (MIRO2)—promotes tumor cell invasion and metastasis through a negative regulation of RhoA. Our previous work identified higher MIRO2 mRNA expression in cancer vs. normal patient samples, which correlated with worse patient outcomes. Furthermore, in the context of prostate cancer we demonstrated that MIRO2 was critical for 2D cell growth, anchorage-dependent and -independent colony formation, and tumor growth in vivo. However, it remains completely unknown if MIRO2 affects primary tumor growth or if this protein is important throughout tumor progression. Using siRNA mediated knockdown (KD) of MIRO2 we clearly show that MIRO2 KD ubiquitously reduces tumor cell invasion in breast, melanoma, pancreatic, and prostate cancer cell lines. Preliminary experiments modeling late-stage metastasis showed that tail vein injection of PC3 cells stably expressing MIRO2 targeting shRNA had reduced kidney and liver metastatic burden in comparison to PC3 cells with control shRNA. In further mechanistic studies, we identified novel MIRO2 binding partners and used siRNA to KD the top hits and observe changes in invasive capacity. We found that atypical myosin IX B (MYO9B) reduced the invasion of cells to the largest extent. MYO9B is a plus-end direct atypical myosin known to control cell motility through spatially inactivating RhoA at the leading edge of migrating cells. Excitingly, MIRO2 KD showed an increase in active RhoA phenocopying MYO9B KD. Lastly, in pilot experiments we found that dual KD of MIRO2 and RhoA rescued invasive capacity in comparison to MIRO2 KD alone. Taken together, we propose a novel signaling mechanism by which MIRO2 broadly promotes invasion and metastasis through MYO9B dependent inactivation of RhoA.

Citation Format: Dillon P. Boulton, Madison Furnish, M. Cecilia Caino. A novel role for Mitochondrial Rho GTPase 2 in tumor cell invasion and metastasis [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A043.
Abstract A046: Heterogenous nuclear ribonucleoprotein E1 regulates leukemia inhibitory factor receptor (LIFR) expression through FAM3C in mammary epithelial cells
Authors: Streitfeld William S. , Dalton Annamarie C. , Howe Philip H.
Published: January 2023

Leukemia inhibitory factor receptor (LIFR) signals through signal transducer and activator of transcription 3 (STAT3), a transcription factor that is associated with poor prognosis due to its overexpression and/or hyperactivity in 40% of breast cancer cases. LIFR has been defined as either a promoter or suppressor of metastasis in mammary carcinoma, and recently we have shown that its expression is significantly upregulated following loss of the RNA-binding protein heterogenous nuclear ribonucleoprotein E1 (hnRNP E1, “E1”). We have further shown that E1 is a regulator of the epithelial-to-mesenchymal transition (EMT) and of differentiation in mammary epithelium. Mechanistically, E1 acts as a tumor suppressor by binding to and inhibiting translation of mRNAs involved in EMT and cellular self-renewal, including the cytokine FAM3C. We have demonstrated that FAM3C binds to LIFR in the extracellular space and activates STAT3. Herein, we characterize induction of LIFR expression in response to FAM3C/LIFR signaling, and show that E1 upregulates LIFR transcription through FAM3C, involving initiation at the LIFR promoter locus. Induction of LIFR expression in mammary epithelial cells has been shown to be associated with tumor initiation, breast cancer stem cells, and metastasis. Additionally, our bioinformatic analysis reveals a signature of transcriptional regulation associated with FAM3C/LIFR interaction. Comparison between RNA-sequencing data from our mouse model of mammary carcinoma and human mammary carcinoma cell lines reveals common biological pathways and potential therapeutic targets. Future directions will be aimed at investigating the effect of modulation of LIFR expression during tumor dissemination in vivo.

Citation Format: William S. Streitfeld, Annamarie C. Dalton, Philip H. Howe. Heterogenous nuclear ribonucleoprotein E1 regulates leukemia inhibitory factor receptor (LIFR) expression through FAM3C in mammary epithelial cells [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr A046.
Abstract B009: Tumoral activation of endothelium drives macrophages-mediated metastatic niche formation and promotes lung metastasis
Authors: Chung Hyewon , Kim Sang Wha , Seok Seung Hyeok
Published: January 2023

Primary tumor induces macrophages-enriched environment to establish metastatic niche which enhances homing and colonization of circulating tumor cells. However, the molecular mechanism underlying endothelial remodeling that favors macrophages infiltration and subsequent niche formation is not fully understood. Here, we found that tumor-derived factors activated endothelial cells to increase adhesive efficacy of monocytes to the endothelium, leading to macrophage infiltration. In mouse models of pulmonary metastasis, this endothelial activation triggered formation of premetastatic niche via accumulation of macrophages in the lungs. Of note, macrophages primed the metastatic microenvironment through enhancing expression of niche-related genes including S100A8, S100A9, MMP9 and fibronectin within the premetastatic lungs and directly transmitted survival signal to tumor cells in a contact-dependent manner. Furthermore, we demonstrated that depletion of macrophages specifically during premetastatic stages reduced niche formation and also resulted in reduced metastatic burden. These findings suggest that endothelial remodeling at metastatic site is a key step for initiation of premetastatic niche formation supported by macrophages. Targeting this step could present an opportunity for therapeutic intervention of metastatic spread in patients with malignant cancers.

Citation Format: Hyewon Chung, Sang Wha Kim, Seung Hyeok Seok. Tumoral activation of endothelium drives macrophages-mediated metastatic niche formation and promotes lung metastasis [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr B009.
Abstract B036: Platelets promote in vitro osteosarcoma cell migration and in vivo metastatic lung seeding
Authors: Travis Olivia K. , Wang Ruishan , Labelle Myriam
Published: January 2023

Osteosarcoma (OS) is the most common primary bone cancer worldwide where it mainly affects adolescents. Metastatic OS has a 20% survival rate and treatment options remain poor, thus understanding mechanisms that contribute to OS metastasis is critical for improved patient outcomes. Platelets, commonly recognized for their role in hemostasis, have been shown to promote metastasis in several cancer models. However, most previous studies investigating platelet contributions to metastasis have focused on carcinomas while their role in osteosarcoma remains poorly understood. Previous studies by our lab have shown that platelet interactions with breast cancer cells resulted in increased migration in vitro and increased lung metastases in vivo. Thus, we hypothesized that OS cell migration will be enhanced in the presence of platelets, and that platelet depletion in vivo will inhibit OS lung seeding. Cell suspensions of SaOs2 and MG63.3 cells in serum free media were fluorescently stained and seeded, in triplicates, in the top chamber of an 8um pore Fluoroblock migration assay wells. In the bottom chamber, media and either Tyrode’s buffer or 1.5x106 isolated human platelets were added. The presence of platelets resulted in a 3.5- and 4.6-fold increase of migrated MG63.3 and SaOs2 cells respectively. For in vivo analysis, cohorts of eight-week-old, male Nod Scid-Gamma (NSG), Nod Scid, and Nude mice were randomly assigned to either the control or platelet depletion group. Mice were depleted of platelets by administration of Anti-Platelet glycoprotein Ib alpha (GPIbα) antibody and, 5x105 GFP transduced MG63.3 cells were injected via tail vein. 24 hrs later, the mice were euthanized, and lungs were collected and microscopically imaged. Platelet depletion in NSG mice resulted in a 48% reduction in MG63.3 lung seeding compared to the isotype control recipient mice. This result was more dramatic in the less immunocompromised Nude and Nod Scid mice with a 61% and 87% reduction of MG63.3 lung seeding in the platelet depleted mice respectively. Finally, RNAseq and was performed on seven OS cell lines co-cultured with isolated mouse platelets, and alterations in gene expression were compared to control cells. Gene Enrichment Set Analysis (GSEA) of the data indicated that platelet co-incubation significantly upregulated pathways related to inflammatory responses, Tumor Necrosis Factor (TNF)-a signaling, and Interleukin (IL)-6 signaling. These results suggest that platelet secreted factors facilitate OS cell migration in vitro while an immune dependent mechanism is involved in the role of platelets in the in vivo lung seeding of OS cells.

Citation Format: Olivia K. Travis, Ruishan Wang, Myriam Labelle. Platelets promote in vitro osteosarcoma cell migration and in vivo metastatic lung seeding [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr B036.
Abstract IA001: Metastasis initiating cells and ecosystems
Authors: Massagué Joan
Published: January 2023

Metastasis is initiated and sustained through therapy by cancer cells with stem-like properties, termed metastasis initiating cells (MICs). Recent work suggests that MICs are malignant cells that adopted a regenerative stem-cell phenotype with intrinsic programs to survive the physical, immune and metabolic stresses of the metastatic process, enter a slow-cycling state for dormancy, and establish supportive interactions with organ-specific ecosystems for eventual outgrowth and recurrence. Notably, relapse frequently develops from disseminated MICs that remain dormant in distant organs after the apparently successful treatment of a primary tumor. During dormancy, MICs fluctuate between immune evasive quiescent and cell cycle reentry states, which exposes them to elimination by the immune system. By combining novel immunocompetent mouse models of lung adenocarcinoma indolent metastasis, in vivo genetic screens, immune profiling, and pre-clinical anti-tumor drug testing, we identified specific pathways in reawakened MICs that drive immune-mediated clearance and can be therapeutically activated to hasten the elimination of residual disseminated disease. Moreover, analysis of brain metastasis with advanced imaging reveals that the interplay between MICs and their host ecosystem is influenced by the architecture of metastatic colonies. Thus, lung adenocarcinoma and triple-negative breast cancer metastases grow by spreading on the basement membrane of brain capillaries, forming perivascular tumor colonies that closely interact with microglia and astrocytes. In contrast, HER2+ breast cancer cells adopt a spheroidal growth pattern that is driven by tumor-derived extracellular matrix and restricts microglia and astrocyte infiltration. The use of single-cell analytics revealed distinct microglia activation states, MIC dependencies and vulnerabilities associated with these different growth patterns. This progress sheds light on the complex and dynamic interplay between MICs and their host ecosystems and point at therapeutically actionable targets to hasten the elimination of disseminated disease.

Citation Format: Joan Massagué. Metastasis initiating cells and ecosystems [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr IA001.
Abstract IA006: EMT-induced tumor plasticity in cancer metastasis
Authors: Mani Sendurai A
Published: January 2023

Advancements in diagnosis and treatment help save cancer patients if the patient presents with local disease. However, the survival probability drops significantly when the tumor metastasizes to other organs. Cancer stem cells (CSCs) are responsible for developing resistance to chemotherapy, tumor relapse, and metastasis. We and others found that the latent embryonic program epithelial to mesenchymal transition (EMT), known to induce mesenchymal properties, including increased migration and invasion, can also induce stem cell properties in cancer cells. This finding postulates that the tumor is inherently capable of generating CSCs de novo and promoting plasticity and heterogeneity. Using syngeneic orthotopic tumor models, we found that CD8+ T cells play a vital role in regulating metastasis besides plasticity in tumor cells. This finding suggests that targeting EMT-signaling pathways and immune cells may provide a window of opportunity to treat metastasis, chemoresistance, and tumor relapse.

Citation Format: Sendurai A Mani. EMT-induced tumor plasticity in cancer metastasis [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr IA006.