bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3495: A novel Mediterranean dietary intervention for prostate cancer
Authors: Cass Samuel , Wargo Jennifer , Pettaway Curtis , Pisters Louis , Davis John , Chapin Brian , Ward John , Daniel Carrie , Gregg Justin
Published: June 2022

Active surveillance (AS) is a safe management strategy for men with low-risk prostate cancer; however, nearly 35% of men on AS will undergo definitive treatment within 4 years, most commonly due to disease progression. Determining a non-invasive means of limiting disease progression would allow men to avoid quality of life issues associated with radical prostate treatment. Some modifiable factors, such as obesity and lipid levels, are associated with prostate cancer risk, though no behavioral intervention has been shown to affect prostate cancer outcomes. Using a prospective study of men on active surveillance, our group recently demonstrated that higher quality diets were associated with a significantly lower risk of disease progression. The Mediterranean diet (MD) specifically may be beneficial for men with localized prostate cancer (PCa) on active surveillance (AS) because of its anti-inflammatory, antilipidemic, and chemopreventive properties. Furthermore, recent evidence supports that dietary interventions can induce modulation of the gut microbiome, and consequently, immune function and inflammatory pathways, which warrants further investigation of the microbiome’s role in the context of prostate cancer. Here, we describe a novel dietary intervention among a group of men with localized prostate cancer who are electing to undergo prostatectomy for prostate cancer treatment. The intervention is a feeding study involving the provision of an isocaloric, high quality diet based on the lipid-lowering and anti-inflammatory parameters of the Mediterranean diet, including a strong focus on fiber-rich plant foods and healthy sources of mono and poly-unsaturated fatty acids. The primary outcome in this pilot study is the feasibility of a neoadjuvant dietary intervention. Secondary outcomes include studying the effects of the diet on metabolic parameters, the fecal microbiome, and changes in circulating metabolites shown to be associated with progression on active surveillance. Furthermore, by focusing on African-American and non-Hispanic White males, this study may provide insights into the key differences in inflammatory, immune and microbiome signatures, which may underly the prominent cancer-racial disparity seen in African-American men. Therefore, this study will gain preliminary data needed to inform further dietary interventions that may impact risk of progression in select men with prostate cancer enrolled on active surveillance.

Citation Format: Samuel Cass, Jennifer Wargo, Curtis Pettaway, Louis Pisters, John Davis, Brian Chapin, John Ward, Carrie Daniel, Justin Gregg. A novel Mediterranean dietary intervention for prostate 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 3495.
Abstract 3504: QL415, a tumor targeted IL-15 fusion protein stimulating both lymphoid and myeloid immune cells for optimal anti-tumor immune response
Authors: Tang Irene , Schwimmer Lauren , Jin Monica , Gu Shenda , Prior Wei Wei , Capacio Arianne , Tran Hieu V. , Chan Allan , McClain Anna , Chen Shihao , Cao Chuanzeng , Wu Xiaoran , Xu Yanling , Guan Dongmei , Chen Xi , Su Xianli
Published: June 2022

Cytokines are potent stimulators of the immune system and have long been investigated and used as therapeutics to treat cancer. However, free cytokines have short half-life and the lack of tumor targeting often results in systemic toxicity. IL-15 is a stimulatory cytokine that shares the common beta and gamma receptors with IL-2. In contrast to IL-2, IL-15 is more selective for NK and effector memory T cells, with less proliferative effect on Tregs, making it a more preferred therapeutic candidate for cancer. QL415 is an anti-PD-L1 x IL-15 fusion protein designed to enhance tumor accumulation and prolong blood circulation, thereby widening the therapeutic window. IL-15 and IL-15 receptor alpha sushi domain are covalently linked to the C-terminus of our proprietary PD-L1 antibody. Fc-mediated effector functions were ablated using mutations in the CH2 domain, to prevent depletion of effector immune cells. In vitro studies showed QL415 to be highly potent in promoting proliferation and pSTAT5 signaling of IL-15 -responding NK92 and M07e cells. QL415 selectively induced proliferation of NK and CD8+ T cells from human PBMC in vitro and mouse CD8+ and NK1.1 cells in vivo. In a MC38 mouse colon cancer model expressing human PD-L1, QL401 was highly effective in suppressing tumor growth, in contrast to PD-L1 monoclonal antibody or a non-targeted IL-15 control. Follow up tumor rechallenge study showed protective memory against cognate MC38 but not B16F10. In a separate MC38 tumor model using a mouse surrogate molecule of QL415, immunophenotyping of antigen presenting cells offered preliminary evidence of enrichment of conventional dendritic cells 1 in the tumor draining lymph nodes. Therefore, QL415 not only directly stimulates effector immune cells, but also indirectly through antigen presenting cells, promotes robust anti-tumor response. In non-human primates, Q415 was tolerated up to 1.5 mg/kg, well above the effective therapeutic dose. In light of these favorable preclinical efficacy and safety results, a phase 1 dose escalation and expansion study has been initiated to evaluate the safety, tolerability, and early efficacy of QL415.

Citation Format: Irene Tang, Lauren Schwimmer, Monica Jin, Shenda Gu, Wei Wei Prior, Arianne Capacio, Hieu V. Tran, Allan Chan, Anna McClain, Shihao Chen, Chuanzeng Cao, Xiaoran Wu, Yanling Xu, Dongmei Guan, Xi Chen, Xianli Su. QL415, a tumor targeted IL-15 fusion protein stimulating both lymphoid and myeloid immune cells for optimal anti-tumor immune response [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 3504.
Abstract 3531: Sensitizing immune unresponsive colorectal cancers to immune checkpoint inhibitors through MAVS overexpression
Authors: Hwang Bin-Jin , Tsao Li-Chung , Acharya Chaitanya , Trotter Timothy , Agarwal Pankaj , Wang Tao , Wei Junping , Yang Xiao-Yi , Lei Gang-jun , Osada Takuya , Lyerly Herbert Kim , Morse Michael A. , Hartman Zachary
Published: June 2022

Background: The majority of colorectal carcinomas (CRCs) are insensitive to anti-PD-1/PD-L1 immune checkpoint inhibitor (ICI) antibodies. While there are many causes for ICI insensitivity, recent studies suggest that the suppression of innate immunity through a loss of innate immune adaptor expression could be a root cause of this insensitivity and an important factor in the evolution of tumor immunosuppression.

Methods: We interrogated mitochondrial antiviral signaling gene (MAVS) expression through bioinformatics analyses of multiple datasets to validate its suppression in clinical samples. We then engineered MAVS expressing tumor cells and tested its ability to elicit innate and adaptive anti-tumor immunity using both in vitro and in vivo approaches, which we then confirmed using MAVS expressing viral vectors. Finally, we observed that MAVS stimulated PD-L1expression in different types of tumor cells and then assessed the combination of PD-L1 ICI antibodies with MAVS tumor expression in vivo.

Results: MAVS was significantly downregulated in CRCs, but its re-expression could stimulate broad cellular interferon-related responses, in both murine and patient-derived CRCs. In vivo, local MAVS expression elicited significant anti-tumor responses in both immune-sensitive and insensitive CRC models, through the stimulation of an interferon responsive axis that elicited tumor antigen-specific adaptive immunity. Critically, we found that tumor-intrinsic MAVS expression triggered systemic adaptive immune responses that enabled abscopal CD8+ T cell cytotoxicity against distant CRCs. As MAVS also induced PD-L1 expression, we further found synergistic anti-tumor responses in combination with anti-PD-L1 ICIs.

Conclusion: These data demonstrate that intratumoral MAVS expression results in local and systemic tumor antigen-specific T cell responses, which could be combined with PD-L1 ICI to permit effective anti-tumor immunotherapy in ICI resistant cancers. We are further exploring to overcome the ICI insensitivity of other types of solid tumor, with MAVS-delivery strategies other than viral vectors.

Citation Format: Bin-Jin Hwang, Li-Chung Tsao, Chaitanya Acharya, Timothy Trotter, Pankaj Agarwal, Tao Wang, Junping Wei, Xiao-Yi Yang, Gang-jun Lei, Takuya Osada, Herbert Kim Lyerly, Michael A. Morse, Zachary Hartman. Sensitizing immune unresponsive colorectal cancers to immune checkpoint inhibitors through MAVS overexpression [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 3531.
Abstract 3538: A CTLA-4 targeted ETB for Treg depletion shows favorable preclinical efficacy and safety
Authors: Sarkar Asis , Martin Rebecca , Byrne Lauren R. , Howard Caleigh , Khanna Swati , Rabia Lilia A. , Adhikari Diana , Sousares Michaela M. , Aldana Alvaro , Robinson Garrett L. , Zhao Jay , Moore Chris B. , Iberg Aimee
Published: June 2022

CTLA-4-targeted engineered toxin bodies (ETBs) are designed to deplete immune-suppressive regulatory T cells (Tregs) in the tumor microenvironment (TME) directly and in a manner independent of Fc-mediated effector functions, offering a unique approach to CTLA-4 targeted therapy that may show efficacy where blocking antibodies have failed. ETBs are large molecule proteins consisting of an antibody fragment genetically fused to a proprietary de-immunized (DI) form of the Shiga-like toxin-1 subunit A (SLTA). Once engaged to the specific cell surface target of interest, ETBs internalize, route to the cytosol, and permanently inactivate ribosomes through an irreversible enzymatic process. This results in cell death via apoptotic mechanisms.

Here we describe the preclinical characterization of a candidate CTLA-4-targeted ETB. The candidate ETB kills gain-of-function model cell lines that express CTLA-4 at levels observed on primary human tumor Tregs. Notably, the potency of the candidate ETB is limited when CTLA-4 is expressed at low levels, thereby allowing a mechanism to spare CD8+ cytotoxic T lymphocytes (CTLs) and effector CD4+ T cells from targeted depletion. The candidate ETB binds CTLA-4 with high affinity utilizing a unique biparatopic binding domain. Using in vitro systems, we show that the candidate ETB can block CTLA-4:B7 interactions and repress Treg-mediated T-cell suppression. In a syngeneic mouse pharmacodynamic model, the candidate ETB depletes tumor-resident Tregs and increases the CD8:Treg ratio in the TME. A toxicological study in naive NHP demonstrate that the candidate ETB, which is cross-reactive to NHP, is well-tolerated at 450 mcg/kg (highest dose tested) administered intravenously once weekly for 4-weeks and does not significantly alter the CTLA-4 low/null peripheral T cell populations. Overall, the candidate ETB is a unique CTLA-4 targeting modality that is designed to deplete Tregs selectively and directly in the tumor, spare CD8+ CTLs, and represents an alternative treatment option to traditional blocking antibodies. Development activities are underway to support the upcoming IND application.

Citation Format: Asis Sarkar, Rebecca Martin, Lauren R. Byrne, Caleigh Howard, Swati Khanna, Lilia A. Rabia, Diana Adhikari, Michaela M. Sousares, Alvaro Aldana, Garrett L. Robinson, Jay Zhao, Chris B. Moore, Aimee Iberg. A CTLA-4 targeted ETB for Treg depletion shows favorable preclinical efficacy and safety [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 3538.
Abstract 3558: A novel and versatile cytokine empowered DNA vaccine platform with superior immune activating potential
Authors: Beraas Audun , Dillard Pierre , Wong Judith Jing , Wold Christian Winther , Solbakken Eirik , Oslen Linn Guro , Heim Joel Benjamin , Chellappa Stalin , Fredriksen Agnete , Pedersen Mikkel Wandahl , Granum Stine
Published: June 2022

Immunotherapy, in particular Immune Checkpoint Blockade (ICB) has been regarded as the next standard of care for many solid tumor indications. Yet, a significant proportion of patients do not respond to therapy. More specifically, failure of ICB has been linked to the absence of an anti-tumor immune response that can be boosted or revitalized. Cancer vaccines that are designed to elicit such response by educating the host immune system to recognize tumor antigens, are therefore considered a key next generation therapeutic modality for the treatment of human cancers.

Various vaccine platforms have been developed over the years, but recent advances in delivery technologies combined with the intrinsic qualities of the DNA matrix have positioned DNA vaccines as a safe and flexible alternative to other types of vaccine technologies. Vaccibody is developing DNA vaccines that allows specific targeting of tumors antigens to Antigen Presenting Cells (APCs), thus maximizing the elicited immune response.

Here we present a second-generation version of our DNA platform in which our vaccibody molecule can be co-expressed with immune-stimulatory proteins from one plasmid using a multicistronic design. Compared to the vaccibody molecule alone the simultaneous expression of selected immune stimulatory cytokines was shown to boost the overall immune response almost 3-fold to drive a potent anti-tumor response.

These data demonstrate the flexibility and potential of DNA vaccines as well as the advantages of combining an APC targeted delivery of tumor specific antigens together with a local production of immune stimulatory proteins.

Citation Format: Audun Beraas, Pierre Dillard, Judith Jing Wong, Christian Winther Wold, Eirik Solbakken, Linn Guro Oslen, Joel Benjamin Heim, Stalin Chellappa, Agnete Fredriksen, Mikkel Wandahl Pedersen, Stine Granum. A novel and versatile cytokine empowered DNA vaccine platform with superior immune activating potential [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 3558.
Abstract 3563: CD8+ T cell-mediated tumor rejection by an Adeno-associated virus-like particle (AAVLP) vaccine
Authors: Neukirch Lasse , Schmidt Patrick , Zörnig Inka , Jäger Dirk , Uhrig-Schmidt Silke
Published: June 2022

The utilization of adeno-associated viruses (AAVs) as vaccines has primarily been focused on the induction of antibody responses. Displaying antigens derived from HER2 or HPV on the AAV capsid surface lead to strong humoral immune responses. However, the potential of AAVs as T cell inducing vaccines has been poorly investigated. Model antigens have been inserted into the capsid of AAVs to analyze T cell responses, but these studies rather focused on preventing vector immunity for an improved tolerance of gene therapies. By using this as a starting point, we set out to test AAVs as T cell inducing vaccines with the prospect of targeting cancer neoantigens. In first experiments, the general properties of the vaccination strategy were estimated by displaying the ovalbumin-derived model antigen SIINFEKL on the surface of adeno-associated virus-like particles (AAVLPs). Upon injection of the AAVLP-SIINFEKL vaccine, mice developed strong CD8+ T cell responses against the displayed antigen. Highest immune responses were achieved by subcutaneous hock injection of AAVLPs, adjuvanted by Montanide ISA 51 VG. SIINFEKL-specific T cell responses peaked around three weeks after vaccination, whereas a memory subset remained present for long term. The anti-tumor efficacy of the vaccine was shown by injecting SIINFEKL-expressing B16F10 melanoma cells subcutaneously into mice, in which the tumor was completely rejected after vaccination. Interestingly, the induction of CD8+ T cell responses and the tumor protection depended on the presence of CD4+ T cells. Accordingly, T helper epitopes were identified in the AAVLP capsid sequence. In addition to the initial tests, a set of murine neoantigens, derived from B16F10 cells, were displayed on AAVLPs. In direct comparison to a peptide vaccine, which did not have an effect on tumor growth, the AAVLP vaccine had a significant impact on the tumor growth rate. In conclusion, AAVLPs show promising effects as T cell vaccines. The vaccination strategy can be used to induce cytotoxic T cell responses in general and anti-tumor effects in particular. AAV capsid-specific helper epitopes is one inherent advantage, since the AAVLP vaccine delivers immune stimulation within the viral particle itself. Thus, antigen-displaying AAVLPs could be an alternative to current gold standards in the field of neoantigen vaccines and have a prospect for future clinical applications.

Citation Format: Lasse Neukirch, Patrick Schmidt, Inka Zörnig, Dirk Jäger, Silke Uhrig-Schmidt. CD8+ T cell-mediated tumor rejection by an Adeno-associated virus-like particle (AAVLP) vaccine [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 3563.
Abstract 3580: Integrative genomics of checkpoint blockade response in advanced non-small cell lung cancer
Authors: Ravi Arvind , Gainor Justin , Arniella Monica , Stewart Chip , Freeman Sam , Awad Mark M. , Forde Patrick , Anagnostou Valsamo , Henick Brian , Riess Jonathan W. , Gibbons Don , Pennell Nathan , Velcheti Vamisdhar , Leshchiner Ignaty , Kim Jaegil , Digumarthy Subba , Mino-Kenudson Mari , Heymach John , Vokes Natalie , Griffin Andrew , Ricciuti Biagio , Rizvi Naiyer , Herbst Roy , Velculescu Victor , Brahmer Julie , Schalper Kurt , Janne Pasi , Wolchok Jedd , Shaw Alice , Hacohen Nir , Getz Gad , Hellmann Matthew D.
Published: June 2022

The introduction of checkpoint blockade therapy, specifically anti-PD-1/PD-L1 agents, has transformed the treatment landscape of advanced Non-Small Cell Lung Cancer (NSCLC). While our understanding of the biology underlying immunotherapy in NSCLC is still incomplete, studies to date have established central roles for Tumor Mutation Burden (TMB) and PD-L1 Tumor Proportion Score (PDL1-TPS). In order to expand our understanding of the molecular features underlying response in NSCLC, we describe here the first joint analysis of the Stand Up 2 Cancer-Mark Foundation (SU2C-MARK) Cohort, a collection of 393 patients with whole exome and/or RNA sequencing along with matched checkpoint blockade response annotation. We identify a number of significant associations between molecular features and response, including: 1) favorable and unfavorable genomic subgroups; 2) distinct immune infiltration signatures associated with wound healing (unfavorable) and immune activated (favorable) microenvironments; and 3) a novel de-differentiated tumor-intrinsic subtype characterized by high TMB, immune activation, and enhanced response rate. Taken together, results from this cohort extend our understanding of NSCLC-specific predictors, providing a rich set of molecular and immunologic hypotheses with which to further our understanding of the biology of checkpoint blockade in NSCLC.

Citation Format: Arvind Ravi, Justin Gainor, Monica Arniella, Chip Stewart, Sam Freeman, Mark M. Awad, Patrick Forde, Valsamo Anagnostou, Brian Henick, Jonathan W. Riess, Don Gibbons, Nathan Pennell, Vamisdhar Velcheti, Ignaty Leshchiner, Jaegil Kim, Subba Digumarthy, Mari Mino-Kenudson, John Heymach, Natalie Vokes, Andrew Griffin, Biagio Ricciuti, Naiyer Rizvi, Roy Herbst, Victor Velculescu, Julie Brahmer, Kurt Schalper, Pasi Janne, Jedd Wolchok, Alice Shaw, Nir Hacohen, Gad Getz, Matthew D. Hellmann. Integrative genomics of checkpoint blockade response in advanced non-small cell lung 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 3580.
Abstract 360: Methyl-cantharidimide (MCA) can induce apoptosis by activating UNC5B-Netrin-1-DAPK pathway in hepatocellular carcinoma
Authors: Li Yi-Dong , Mao Yong , Dong Xing-Duo , Lei Zi-Ning , Yang Yuqi , Lin Lizhu , Yang Dong-Hua , Chen Zhe-Sheng
Published: June 2022

In this study, we investigated the underlying mechanism of action of methyl-cantharidimide (MCA), a cantharidin (CTD) analog, as an anticancer drug, in resistant cancer cells overexpressing either ABCB1 or ABCG2 and in cisplatin-resistant cancer cells. The results indicated that: (i) MCA was efficacious in the ABCB1-overexpressing cell line, KB-C2, the ABCG2-overexpressing cell line, NCI-H460/MX20 and in the cisplatin resistant cancer cell lines, KCP-4 and BEL-7404/CP20; (ii) MCA induced apoptosis in both BEL-7404 and BEL-7404/CP20 cancer cells and arrested both BEL-7404 and BEL-7404/CP20 cancer cells in the G0/G1 phase of the cell cycle; (iii) MCA upregulated the expression level of the protein, unc-5 netrin receptor B (UNC5B) in HepG2 and BEL-7404 cancer cells. (iv) MCA can activate UNC5B-Netrin-1-DAPK apoptosis pathway in hepatocellular carcinoma cells. Overall, our results indicated that MCA's efficacy in multiple cancer cell lines is due to the activation of UNC5B-Netrin-1-DAPK apoptosis pathway and cell cycle arrest in the G0/G1 phase.

Citation Format: Yi-Dong Li, Yong Mao, Xing-Duo Dong, Zi-Ning Lei, Yuqi Yang, Lizhu Lin, Dong-Hua Yang, Zhe-Sheng Chen. Methyl-cantharidimide (MCA) can induce apoptosis by activating UNC5B-Netrin-1-DAPK pathway in 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 360.
Abstract 3615: TREM2+ macrophages mediate immunosuppression in tumors
Authors: Molgora Martina , Luccia Blanda Di , Khantakova Darya , Jaeger Natalia , Czepielewski Rafael Sanguinetti , da Silva Cristiane Secca , Bando Jennifer , Gilfillan Susan , Cella Marina , Schreiber Robert , Colonna Marco
Published: June 2022

Checkpoint immunotherapy unleashes T cell control of tumors, but is undermined by immunosuppressive myeloid cells. TREM2 is a myeloid receptor that transmits intracellular signals that sustain microglial responses during Alzheimer’s disease. TREM2 is also expressed by tumor-infiltrating macrophages. Here, we found that Trem2-/- mice are more resistant to growth of various cancers than wild-type mice and are more responsive to anti-PD-1 immunotherapy. Furthermore, treatment with anti-TREM2 mAb curbed tumor growth and fostered regression when combined with anti-PD-1. scRNA-seq revealed that both TREM2 deletion and anti-TREM2 are associated with scant MRC1+ and CX3CR1+ macrophages in the tumor infiltrate, paralleled by expansion of myeloid subsets expressing immunostimulatory molecules that promote improved T cell responses. TREM2 was expressed in tumor macrophages in over 200 human cancer cases and inversely correlated with prolonged survival for two types of cancer. Thus, TREM2 might be targeted to modify tumor myeloid infiltrates and augment checkpoint immunotherapy.

Citation Format: Martina Molgora, Blanda Di Luccia, Darya Khantakova, Natalia Jaeger, Rafael Sanguinetti Czepielewski, Cristiane Secca da Silva, Jennifer Bando, Susan Gilfillan, Marina Cella, Robert Schreiber, Marco Colonna. TREM2+ macrophages mediate immunosuppression in tumors [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 3615.
Abstract 3640: Adipocyte age dependent metabolic programming dictates melanoma metastatic burden and tropism
Authors: Gurung Shilpa
Published: June 2022

The aged skin microenvironment and aged fibroblasts contribute to melanoma progression. A key hallmark of skin ageing is the loss of subcutaneous fat. Because previous work has established that lipids contribute to melanoma invasion and proliferation, we studied whether aged adipocytes contribute to the poor outcome of aged patients. We compared the distinct lipid secretory profile of young and aged human subcutaneous adipocytes, and discovered young and aged adipocytes contribute different lipids to the melanoma microenvironment, activating aged-dependent signaling pathways and driving distinct metabolic programms in melanoma cells.

To test how aged and young adipocytes affect melanoma, we obtained human preadipocytes from donors of different ages and differentiated them to functional adipocytes in vitro. We studied the morphological, transcriptomic, proteomic, and lipid profile of adipocytes by age, then exposed melanoma cells to the young or aged adipocyte secretome. We observed melanoma cells exposed to young adipocyte secretome increased the rate of proliferation, invasion, and migration compared to melanoma cells exposed to aged adipocyte secretomes. This effect was linked to a greater amount of total lipids secreted by young adipocytes, as well as a unique ratio of lipids by age, which were taken up by melanoma cells. Critically, lipid transfer led to distinct signaling pathway activation and metabolic reprogramming. In vivo, these differences led to a greater rate of metastasis and distinct tropism of melanoma cells exposed to aged or young adipocyte secretomes. Finally, we systematically tested the contribution of differentially secreted lipids by age and identified the specific lipids that function as signaling molecules to switch metabolic dependencies and fuel proliferation, invasion, and migration of melanoma cells

Citation Format: Shilpa Gurung. Adipocyte age dependent metabolic programming dictates melanoma metastatic burden and tropism [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 3640.
Abstract 3646: Distinct functions of pancreatic stellate cells in PDAC stromal evolution
Authors: Onate M. Kathrina Casapao , Schleisman Matthew , Xia Zheng , Sherman Mara H.
Published: June 2022

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers in the United States with a five-year relative survival rate of 10.8%. After surgery, radiation and chemotherapy are the next lines of therapy. However, these treatments have not significantly improved clinical outcomes due in part to its unique and characteristic desmoplasia, which limits treatment efficacy. Cancer-associated fibroblasts (CAFs) have been shown to contribute in PDAC’s stromal reaction in part viasecretion of extracellular matrix (ECM) components and growth factors, suggesting that CAFs are pro-tumorigenic. This understanding led to several CAF ablation preclinical studies; surprisingly, these studies demonstrated tumor-suppressive potential for CAFs, highlighting the functional heterogeneity among CAFs in PDAC. This emphasizes the need for greater developmental understanding of this influential cell population. Lineage tracing via our recently established Fabp4-Cre;Rosa26mTmG mouse model revealed that pancreatic stellate cells (PSCs), thought to be the main cellular origin of PDAC CAFs, contribute to a minor faction of CAFs yet appear to have non-redundant functions to alternatively-derived CAFs. Here we delve further into the specific functions of PSC-derived CAFs via single-cell RNA sequencing of pancreatic stellate cells in healthy and tumor pancreas with our Fabp4-Cre;Rosa26mTmG mouse model. To our surprise, initial scRNA-seq analysis illustrated that quiescent pancreatic stellate cells from healthy tissue appear to be more heterogeneous to PSC-derived CAFs, with quiescent pancreatic stellate cells clustering into three “super clusters” while PSC-derived CAFs clusters into one “super cluster”. Additionally, clusters specific to normal pancreas tissue revealed potential transcripts-of-interest that are anti-tumorigenic. Current efforts seek to analyze stromal evolution with respect to CAF cell of origin in pre-malignant lesions and throughout stepwise tumorigenesis to metastasis. Furthermore, we are investigating the functional significance of potentially anti-tumorigenic factors lost from the PSC compartment upon differentiation to a CAF phenotype during PDAC progression. These findings shed light on the increasingly appreciated heterogeneity of PDAC CAFs, whilst suggesting that PSC-derived CAFs may be specifically targeted as a singular cell population for therapeutic treatment. Importantly, these findings help gain a greater understanding of PDAC disease progression and development in order to better serve patient care.

Citation Format: M. Kathrina Casapao Onate, Matthew Schleisman, Zheng Xia, Mara H. Sherman. Distinct functions of pancreatic stellate cells in PDAC stromal evolution [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 3646.
Abstract 3648: Hedgehog represses angiogenesis in PDAC through a paracrine cascade mediated by Wif1
Authors: Hasselluhn Marie C. , Decker Amanda R. , Garcia Alvaro Curiel , Maurer Carlo , Thomas Dafydd H. , Olive Kenneth P.
Published: June 2022

Pancreatic Ductal Adenocarcinoma (PDAC) is characterized by an extremely heterogeneous tumor microenvironment (TME) paired with a sparse and compromised vascular network. These features of the TME shape drug responses and contribute to PDAC’s distinctive chemotherapeutic resistance. Prior work demonstrated that inhibition of the Hedgehog (Hh) signaling pathway using Smoothened (Smo) inhibitors led to increased angiogenesis. However, the precise mechanisms through which the Hh pathway regulates angiogenesis has remained elusive and subsequent studies with different agents and timepoints yielded conflicting data. Frustratingly, the phenotype could not be recreated in simple co-culture models.

In order to maintain the cellular heterogeneity and spatial organization that characterizes PDAC, we developed a novel tumor explant system for short-term cultures of both murine and human PDAC. Briefly, we cultured intact 300 μm slices of PDAC tissue on a modified gelatin platform system with a novel culture medium informed by the metabolic composition of PDAC tumor interstitial fluid. This system replicates some of the nutrient/waste gradient of PDAC and enables tumors to be cultured up to one week maintaining the representation of multiple cell types. Studies in explants were complemented by work in the Kras-LSL.G12D/+, p53-LSL.R172H/+, Pdx1-Cre (KPC) genetically engineered model of PDAC.

Comparison of short versus long-term Smo inhibition in KPC mouse pancreatic tumors revealed a robust increase in vessel density in short-term treatment that is lost at later timepoints. Studies in murine and human PDAC explants employing different Smo inhibitors demonstrated increased endothelial tip cell formation upon two days of treatment, resulting in increased vessel density after four days. Computational analyses of human PDAC single cell RNAseq data identified candidate cellular subsets potentially involved in the signaling cascade. RNAseq of pre- and on-treatment tumor samples from KPC mice identified Wif1 as a candidate Hh pathway target gene that effected these programs. Mechanistic studies in tumor explants reveal a complex, multi-step mechanism involving several cell types that led to suppression of angiogenesis in PDAC. These studies both provide a candidate mechanism for Hh-mediated angiosuppression and also demonstrated how the tumor explant platform can elucidate complex intercellular signaling cascades in both murine and human PDAC.

Citation Format: Marie C. Hasselluhn, Amanda R. Decker, Alvaro Curiel Garcia, Carlo Maurer, Dafydd H. Thomas, Kenneth P. Olive. Hedgehog represses angiogenesis in PDAC through a paracrine cascade mediated by Wif1 [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 3648.
Abstract 3652: Maternal body mass index, diabetes, and gestational weight gain and risk for pediatric cancer in offspring: A systematic review and meta-analysis
Authors: Marley Andrew R. , Domingues Allison , Ghosh Taumoha , Turcotte Lucie M. , Spector Logan G.
Published: June 2022

Introduction: Pediatric cancer incidence has steadily increased concurrent with the rise in adult obesity, however, associations between maternal obesity and associated comorbidities and pediatric cancer risk remain understudied.

Methods: A comprehensive and systematic literature search in PubMed and EMBASE databases from their inception to March 15th, 2021. Eligible studies reported risk estimates, sample sizes, and provided sufficient description of outcome and exposure ascertainment. Studies were excluded if not complete, published, peer-reviewed studies, or if provided incompatible exposure data. Data quality was assessed according to Newcastle-Ottawa Scale (NOS) guidelines and reported according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Random-effects models were used to estimate pooled effects.

Results: Thirty-four studies were included, covering 3,404,747 subjects and 14,706 pediatric cancer cases for pre-pregnancy BMI; 14,748,772 subjects and 44,628 pediatric cancer patients for maternal diabetes; and 2,124,647 participants and 15,915 pediatric cancer cases for gestational weight gain. Pre-pregnancy BMI was significantly associated with leukemia risk in offspring (OR per 5-unit BMI increase =1.07 [95% CI = 1.04-1.11] I2 = 0.0%). Any maternal diabetes was positively associated with acute lymphoblastic leukemia risk (OR=1.46 [95% CI = 1.28-1.67] I2 = 0.0%), even after restricting to birthweight-adjusted analyses (OR [95% CI] = 1.74 [1.29-2.34] I2 = 0.0%), and inversely associated with risk of central nervous system tumors (OR=0.73 [95% CI = 0.55-0.97] I2 = 0.0%). Pre-gestational diabetes (OR=1.57 [95% CI = 1.11-2.24] I2 = 26.8%) and gestational diabetes (OR=1.40 [95% CI = 1.12-1.75] I2 = 0.0%) were also significantly associated with acute lymphoblastic leukemia risk. No significant associations were observed for gestational weight gain and pediatric cancer risk.

Conclusions: Maternal obesity and diabetes may be etiologically linked to pediatric cancer, particularly leukemia and central nervous system tumors. Our findings support appropriate weight management and glycemic control as important components of maternal care and offspring health. Further validation studies are warranted.

Citation Format: Andrew R. Marley, Allison Domingues, Taumoha Ghosh, Lucie M. Turcotte, Logan G. Spector. Maternal body mass index, diabetes, and gestational weight gain and risk for pediatric cancer in offspring: A systematic review and meta-analysis [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 3652.
Abstract 3656: Breast cancer care quality in Peru
Authors: Anampa-Guzman Andrea , Perez-Mendez Rushmely Gaby , Huaccho-Rojas Juan , Villar-Lopez Martha , Mini-Diaz Elsy
Published: June 2022

Although many studies indicate that the use of complementary medicine by cancer patients is common and widespread, few studies focused on patients' quality of care using complementary therapies. The overall health care quality perception of patients in Lima, the capital of Peru, was reported to be 68.9% according to the SERVQUAL questionnaire, which grades as acceptable. Breast cancer is the most common cancer in Peru. This study aims to determine the perception of health care quality for breast cancer patients in Lima. This is a cross-sectional study. The perspectives of 182 breast cancer patients about the quality of care at the largest hospital in Lima were assessed. One hundred seventeen patients (64.29%) were only seen at the Oncology Department, and 65 patients (35.71%) were seen at both the Oncology Department and Complementary Medicine Department. The quality perception is based on five dimensions: Reliability, responsiveness, assurance, empathy, and tangibility. The percentage has to be 60 to be an acceptable level. The average age was 58.81 years. The overall and oncology-only groups quality perceptions were not acceptable, 57.32%, and 53.65%, respectively. The oncology and the complementary patient group reported adequate general perception (63.92%). Perception of empathy and assurance were at acceptable levels for both groups. Reliability and tangibility did not receive a satisfactory grade in any group. The responsiveness level was only adequate in the complimentary and oncology group. The breast cancer patients’ perception of the health care quality who received Complementary Medicine services was significantly higher than the oncology-only group. We did not find a significant difference between the dimensions of quality and educational level, type of insurance, and the professional who attended the patient. In conclusion, the patients who were seen at the oncology and complementary medicine reported a better perception of the quality of healthcare. More studies are needed to improve healthcare in breast cancer patients.

Citation Format: Andrea Anampa-Guzman, Rushmely Gaby Perez-Mendez, Juan Huaccho-Rojas, Martha Villar-Lopez, Elsy Mini-Diaz. Breast cancer care quality in Peru [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 3656.
Abstract 3688: Survival benefit of adjuvant radiotherapy among women with high comorbidity burden and low-risk breast cancer: SEER-MHOS analysis
Authors: Lee Eunkyung , Hines Robert B. , Zhu Jianbin , Montan Kayla , Rovito Michael J.
Published: June 2022

Background: The National Comprehensive Cancer Network Breast Cancer Guidelines Committee has suggested that adjuvant radiotherapy (RT) after breast-conserving surgery (BCS) could be omitted among older women with low-risk breast cancer (early-stage, node-negative, and ER-positive) if they were treated with endocrine therapy. However, the effect of RT among older women with multimorbidity is not clearly understood.

Objective: We examined the effect of adjuvant RT on survival outcomes among older women. We also assessed whether the association between RT and survival is modified by comorbidity burden.

Methods: A total of 1105 older women (65+) diagnosed with low-risk breast cancer in 1998-2012 and who received BCS were identified from the population-based Surveillance, Epidemiology, and End-Results-Medicare Health Outcomes Survey (SEER-MHOS) data resource. A propensity score-based inverse probability of treatment weight (IPTW) was calculated with weighting applied to Cox regression models to obtain hazard ratios for the association of RT with survival outcomes. Stratified analysis was conducted to examine differential effects according to the comorbidity burden class, which was identified using the latent class analysis.

Results: Over a 5-year follow-up period, a total of 117 deaths occurred (25 cancer-related and 92 noncancer-related). The IPTW-adjusted Cox proportional hazard analysis showed that RT omission did not significantly increase the risk for 5-year cancer-specific deaths (P=0.202). However, RT omission increased the risk for 5-year noncancer death by 1.84-fold (95% CI=1.35, 2.51) and all-cause death by 1.49-fold (95% CI=1.41, 1.95). The stratified analysis showed that the effect of RT omission on the risk of breast cancer-specific deaths was similar across the different comorbidity groups. For noncancer and all-cause deaths, the effect of RT omission varied by comorbidity burden. In Class 1, RT omission was not associated with an increased risk of noncancer and all-cause deaths. However, in Classes 2 and 3, RT omission is significantly associated with an increased risk of 5-year all-cause deaths in a dose-response manner: by 1.66-fold (95% CI=1.07, 2.56) in Class 2 and 1.96-fold in Class 3 (95% CI=1.13, 3.41). Similar results were observed for 10-year survival.

Conclusions: RT omission does not increase the risk of cancer-specific death and appears to be a valid clinical decision for older women with low-risk breast cancer and multimorbidity.

Citation Format: Eunkyung Lee, Robert B. Hines, Jianbin Zhu, Kayla Montan, Michael J. Rovito. Survival benefit of adjuvant radiotherapy among women with high comorbidity burden and low-risk breast cancer: SEER-MHOS analysis [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 3688.
Abstract 3691: Dynamic BH3 profiling identifies novel combinations in malignant pleural mesothelioma with in vivo efficacy
Authors: Potter Danielle S. , Du Ruochen , Chow Kin-Hoe , Ligon Keith L. , Bueno Raphael , Letai Anthony
Published: June 2022

Currently, malignant pleural mesothelioma (MPM) is mainly treated with conventional therapies such as surgery, chemotherapy and radiation, and patients often suffer from the lack of second line therapy options. In this study, we aimed to identify efficacious drug combinations in primary MPM patient samples and novel MPM patient-derived xenograft (PDX) models to provide better therapeutic options for MPM patients. BH3 profiling is a functional assay that measures mitochondrial priming of the cell. It uses BH3 peptides derived from the BH3 domain of pro-apoptotic BH3-only Bcl-2 family members to provoke a response from viable mitochondria. When tumor cells are treated with drugs prior to BH3 profiling, early changes in their apoptosis signaling can be captured before frank apoptosis occurs. This method is termed dynamic BH3 profiling (DBP), which has been applied in multiple cancer types to identify efficacious drugs. Here, we used high throughput dynamic BH3 profiling (HTDBP) to assess hundreds of drug treatments simultaneously to identify drug combinations that increased apoptotic priming in primary MPM cells ex vivo. Next, we created novel MPM PDX models and tested these drug combinations to determine the ones that primed MPM PDX cells. The HTDBP result from MPM PDX cells recapitulated the chemical vulnerabilities of the primary MPM cells. Moreover, one of the top combination candidates that comprised of navitoclax (BCL-xL, BCL-2 and BCL-w antagonist) and AZD8055 (mTORC1/2 inhibitor) was shown to be safe and efficacious in vivo in our CPDM_011x MPM PDX model. Finally, on the molecular level, we revealed that treatment with navitoclax drove the anti-apoptotic dependence of MPM cell lines towards MCL-1, whose level was downregulated by AZD8055 treatment. In this study, we identified a novel drug combination for MPM with HTDBP that consists of navitoclax and AZD8055. In addition, our results further corroborated that DBP, as a functional assay, can be used to rationally construct novel drug combinations to improve treatment outcomes for cancer patients.

Citation Format: Danielle S. Potter, Ruochen Du, Kin-Hoe Chow, Keith L. Ligon, Raphael Bueno, Anthony Letai. Dynamic BH3 profiling identifies novel combinations in malignant pleural mesothelioma with in vivo efficacy [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 3691.
Abstract 3695: Overexpression of HSD17B13 induced G1 arrest in hepatic cell lines detected by anti-HSD17B13 recombinant rabbit monoclonal antibody
Authors: Qu Tianli , Wu Lipeng , Yom Jina , Guo Zhaoying , Hu Xiaomin , Ren Qi , Niu Qian , Lin Ruei-Shiuan , Yin Dezhong , Liu Xuan , Wei Fu
Published: June 2022

Hepatocellular carcinoma (HCC) is one of the most prevalent tumors worldwide. Hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13), which was newly identified as a liver lipid droplet associated protein, was recently found to be down-regulated in HCC. However, the effects of HSD17B13 on the underlying mechanisms of HCC have not yet been fully explored. For this purpose, HSD17B13 gene was knockdown with CRISPER/Cas9 technique in several HCC cell lines. HSD17B13 expression was detected by Flow cytometry and Western blotting with recombinant rabbit monoclonal antibodies (clone OTIR3G2 and OTIR5C10) against HSD17B13 which were screened for use in immunohistochemistry, western blot, and ELISA application. Overexpressed HSD17B13 induces hepatic cell line G1 capture and increases several G1 arrest-related protein expression, including p21, p27, MMP2 and MMP3. Then the proliferation and cell cycle were suppressed. HSD17B13 gene knockdown will reverse the suppression. These data indicated that HSD17B13 play important roles in the progression of HCC lesions. HSD17B13 might be a potential immune-oncology marker for HCC.

Citation Format: Tianli Qu, Lipeng Wu, Jina Yom, Zhaoying Guo, Xiaomin Hu, Qi Ren, Qian Niu, Ruei-Shiuan Lin, Dezhong Yin, Xuan Liu, Fu Wei. Overexpression of HSD17B13 induced G1 arrest in hepatic cell lines detected by anti-HSD17B13 recombinant rabbit monoclonal antibody [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 3695.
Abstract 3702: UBE4B interacts with the ITCH E3 ubiquitin ligase to induce Ku70 and c-FLIPL K63/K48 branched polyubiquitination and enhanced neuroblastoma HDACi-mediated apoptosis
Authors: Clorennec Christophe Le , Subramonian Divya , Huo Yuchen , Zage Peter E.
Published: June 2022

Expression of the UBE4B ubiquitin ligase is associated with neuroblastoma patient outcomes, but the functional roles of UBE4B in neuroblastoma pathogenesis are not known. We evaluated interactions of UBE4B with the E3 ubiquitin ligase ITCH/AIP4 and the effects of UBE4B expression on Ku70, c-FLIPL, and p53 ubiquitination and proteasomal degradation by co-immunoprecipitation and Western blots. We also evaluated the role of UBE4B in apoptosis induced by HDAC inhibition in our neuroblastoma cell lines model using Western blots. UBE4B was found in a complex with ITCH, with binding mediated by WW domains in the ITCH protein. ITCH activation led to ITCH-UBE4B E3-E4 ubiquitin ligase complex formation and recruitment of Ku70 and c-FLIPL via ITCH WW domains, followed by Ku70 and c-FLIPL Lys48/Lys63 branched polyubiquitination and proteasomal degradation. Histone deacetylase (HDAC) inhibition induced Ku70 and c-FLIPL acetylation, leading to release of c-FLIPL and Bax from Ku70, increased Ku70 and c-FLIPL Lys48/Lys63 branched polyubiquitination via the ITCH-UBE4B complex, and induction of apoptosis. UBE4B depletion in our neuroblastoma cell lines model led to reduced polyubiquitination and increased levels of Ku70 and c-FLIPL proteins leading to the massive reduction of apoptosis induced by HDAC inhibition via stabilization of Ku70 and c-FLIPL proteins allowing the inhibition of the full caspase 8 activation. Our results have identified novel interactions and novel targets for UBE4B ubiquitin ligase activity and a direct role of the ITCH-UBE4B complex in responses of neuroblastoma cells to HDAC inhibition, suggesting that the ITCH-UBE4B complex plays a critical role in responses of neuroblastoma to therapy and suggesting a potential mechanism underlying the association of UBE4B expression with neuroblastoma patient outcomes.

Citation Format: Christophe Le Clorennec, Divya Subramonian, Yuchen Huo, Peter E. Zage. UBE4B interacts with the ITCH E3 ubiquitin ligase to induce Ku70 and c-FLIPL K63/K48 branched polyubiquitination and enhanced neuroblastoma HDACi-mediated apoptosis [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 3702.