bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2732: Multiplexed immunohistochemistry and deep learning: a new approach to spatially map the tumor microenvironment
Authors: Noureddine Kouther , Gallagher Paul , Guillaud Martial , MacAulay Calum
Published: July 2021

The usual standard of care for assessing if a patient has cancer, its stage and its likely future biological behavior is visual examination of one or more H&E and/or Immunohistochemistry(IHC) stained sections. The paradigm of digital pathology has changed, moving from single-marker IHC towards multiplexed labeling, increasing the need for more advanced techniques that can be easily integrated in routine clinical pathology. Although recent advances in multiple immunostaining have enabled characterization of several parameters on a single tissue section. For a higher dimensional chromogen based methodology, we have developed a multiplexed IHC procedure combining multiple labels per round, enabling analysis of complex immune cell population's on a single slide through consecutive cycles of staining, destaining, hyperspectral imaging and spectral unmixing of the chromogen biomarkers in each round. Not only does this allow us to completely visualize and spatially map the tumor microenvironment(TME), but through the application of AI we can further recognize common histological features present in the tissue. Robust, accurate, segmentation of cell nuclei for overlapping nuclei is one of the most significant unsolved issues in digital pathology. Analyzing the cell-cell interactions between immune and tumor cells and identifying clinically relevant patterns may improve patient outcomes while informing on the likelihood of success of possible treatments. By combining a multiplexed IHC technique which enables the detection of multiple markers on a single slide with deep learning segmentation methods to segment every individual cell nuclei in tissue sections with an accuracy comparable to human annotation, we can improve the accuracy of tissue classification based upon the measured characteristics of these cells and their spatial organization. The two techniques joined can be scaled up to the entire tissue section level, providing an accurate spatial cell level representation of the tissue.

Citation Format: Kouther Noureddine, Paul Gallagher, Martial Guillaud, Calum MacAulay. Multiplexed immunohistochemistry and deep learning: a new approach to spatially map the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2732.
Abstract 2744: The immune landscape of recurrent ovarian carcinoma
Authors: Cun Han T. , Ferri-Borgogno Sammy , Cheng Rita , Kim Jae-hoon , Han Gwan Hee , Burks Jared K. , Lu Karen H. , Mok Samuel C.
Published: July 2021

By examining the immune landscape in recurrent ovarian carcinoma, we can characterize the evolution of the tumor immune microenvironment (TIME) to understand its disease progression and response to therapy. Thus, the objective of this study is to evaluate the differences in the TIME in primary HGSOC compared to recurrent disease by utilizing imaging mass cytometry (IMC).

Treatment naive primary HGSOC along with matched first recurrence tumors from the same patient were collected under an IRB-approved protocol. Formalin fixed paraffin embedded tissue sections were stained with 18 metal-tagged antibodies to detect various cell specific and immune related markers using the Fluidigm protocol. IMC data was obtained by the Fluidigm Helios CyTOF instrument utilizing the Hyperion Imaging System laser ablation module. Images from a 1mm2 area of each tissue section were processed and converted to tiff files using MCD Viewer (Fluidigm). Multiplex image analysis was performed with Visiopharm software, where tumor and stromal areas were separated based on presence or absence of Keratin 8/18, Collagen-I, and SMA respectively. Cells boundaries were determined by a pretrained Artificial Intelligence algorithm and phenotyped by Visiopharm unbiased autoclustering module using only the top 20% of pixel values per cell. Statistical analysis was performed using wilcoxon signed-rank test between cell densities of the primary and paired recurrent tumors.

Paired primary and recurrent HGSOC samples were assessed from 27 patients. Stage of disease ranged from I to IV, but most were stage III (81.5%). The median time to first recurrence was 28 months (IQR 34). The image analysis showed that recurrent tumors had a lower density of stromal CD8a+ (p<0.01), CD8+granzymeB+CD45RO+, (p<0.01) and CD8+granzymeB+CD44+CD45RO+ cells (p<0.01) compared to primary tumors. Both stromal and intratumoral CD20+ cells were higher in recurrent tumors (p=0.03, p<0.01). Stromal CD73+Coll-I+ cells were lower in recurrent tumors (p<0.01).

Our study demonstrates the evolution of the TIME of HGSOC from primary tumor to first recurrence. Recurrent tumors demonstrated lower stromal CD8+ T cells, specifically activated memory T cells and resting memory T cells, compared to their primary tumors, suggesting a more immunosuppressive environment. Although recurrent tumors had a higher density of CD20+ B cells, their function was not further described. These alterations propose that recurrent HGSOC demonstrates a more suppressive TIME that warrants further characterization.

Citation Format: Han T. Cun, Sammy Ferri-Borgogno, Rita Cheng, Jae-hoon Kim, Gwan Hee Han, Jared K. Burks, Karen H. Lu, Samuel C. Mok. The immune landscape of recurrent ovarian carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2744.
Abstract 2750: Prognostic impact of tumor infiltrating lymphocytes in the tumor microenvironment of vulvar squamous cell carcinoma
Authors: Lennartz Maximilian , Blessin Niclas Christian , Rolschewski Ann-Christin , Simon Ronald , Sauter Guido , Uhlig Ria , Wölber Linn , Mahner Sven , de Gregorio Nikolaus , Klapdor Rüdiger , Kalder Matthias , Steurer Stefan , Wilczak Waldemar , Burandt Eike , Kind Simon
Published: July 2021

The amount of tumor infiltrating lymphocytes (TILs) is an established prognostic feature in the cancer microenvironment of colorectal and breast cancers, but data are less clear for most other solid cancer types. To study the prognostic impact of TILs in vulvar squamous cell carcinomas, a cohort of 530 cancers was analyzed by immunohistochemistry for CD3 and CD8 positive cells (CD3+, CD8+). Automated digital image analysis was used to quantitate the densities of CD3+, and CD8+ TILs in the invasive margin and the tumor center. The average immune cell density was significantly higher in the invasive margin (CD3+: 1772±1105, CD8+: 769±644 cells/mm2) as compared to the center of the tumor (CD3+: 518±570, CD8+: 301±445 cells/mm2, p≤0.0001 each). Comparison with clinic-pathological data revealed a significant prognostic impact of CD3+ but not of CD8+ cell density and a stronger link with tumor phenotype and outcome for TILs at the invasive tumor margin as compared to the tumor center. Elevated densities of CD3+ cells within the invasive margin were significantly associated with a favorable pT stage (p=0.0012) and a prolonged overall (p=0.0025) and progression free survival (p=0.0231). A high CD3+ cell density in the tumor center was linked to overall (p=0.0457) and progression free survival (p=0.0266). Unsupervised clustering of CD3+/CD8+ cell densities identified 3 major subsets corresponding to the immune desert (sparse TILs at invasive margin and tumor center), immune excluded (high TIL numbers at invasive margin but low TIL counts in the tumor center), and immune inflamed phenotypes (high TIL numbers at invasive margins and tumor center). Survival analysis revealed a particularly poor prognosis for the immune desert phenotype for overall (p=0.0071) and progression free survival (p=0.0027). In summary, our data suggest a clinical utility of TIL quantification in vulva squamous cell cancer.

Citation Format: Maximilian Lennartz, Niclas Christian Blessin, Ann-Christin Rolschewski, Ronald Simon, Guido Sauter, Ria Uhlig, Linn Wölber, Sven Mahner, Nikolaus de Gregorio, Rüdiger Klapdor, Matthias Kalder, Stefan Steurer, Waldemar Wilczak, Eike Burandt, Simon Kind. Prognostic impact of tumor infiltrating lymphocytes in the tumor microenvironment of vulvar squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2750.
Abstract 2759: Reprogramming of PD1+ M2-like tumor-associated macrophages with anti-PD-L1 and Lenalidomide in cutaneous T cell lymphoma
Authors: Han Zhen , Su Chingyu , Wu Xiwei , Qin Hanjun , Rosen Steven T. , Querfeld Christiane
Published: July 2021

M2-like tumor-associated macrophages (TAMs) are abundant and influence cutaneous T cell lymphoma (CTCL) development by inducing immunosuppression. Although it is well established that depletion of M2-like TAMs delays CTCL development, little is known about the underlying mechanisms that shape programmed cell death 1+ (PD1+) M2-like TAMs phenotype in CTCL. Here we identified PD1+ M2-like TAMs accumulated in CTCL lesional skin. Moreover, RNA-seq analysis of human CTCL tissues revealed an up-regulation of the TLR/NF-κB and JAK/STAT signaling pathways. In vitro, PD1+ TAMs exhibited an M2-like profile, with a significant increase in the expression of CD163, CD206, and IL-10, and a clear decrease in the expression of CD80, IL-1β, CXCL-10, and CXCL-11, and the activation of TLR/NF-κB and JAK/STAT signaling pathways. To determine whether PD1+ M2-like TAMs could be reprogrammed, anti-PD-L1 (durvalumab) and lenalidomide were used for treatment of MyLa-conditioned media induced human peripheral blood monocyte-derived PD1+ M2-like TAMs. The results show that anti-PD-L1 and lenalidomide synergistically reshaped M2-like TAMs to M1-like TAMs in vitro through ablation of the TLR/NF-κB and JAK/STAT signaling pathways, which was linked with functional changes in phagocytic activity and cell migration. In conclusion, TLR/NF-κB and JAK/STAT signaling pathways may drive PD1+ M2-like TAMs programming in the CTCL tumor microenvironment, anti-PD-L1 and lenalidomide may reshape PD1+ M2-like TAMs and induce functional changes through ablation of these specific pathways in CTCL.

Corresponding author:

Christiane Querfeld, MD, PhD. Div. of Dermatology and Beckman Research Institute, City of Hope. 1500 E. Duarte Road, Duarte, CA 91010. Email: cquerfeld@coh.org Phone: 626-634-4436 Fax: 626-218-6190

Citation Format: Zhen Han1,5, Chingyu Su1,5, Xiwei Wu2,5, Hanjun Qin2,5, Steven T. Rosen3,5, Christiane Querfeld1,3,4,5. Reprogramming of PD1+ M2-like tumor-associated macrophages with anti-PD-L1 and Lenalidomide in cutaneous T cell lymphoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2759.
Abstract 2764: Mechanisms of genomic-microenvironmental interactions in uveal melanoma
Authors: Kaler Christopher J. , Dollar James J. , Kurtenbach Stefan , Kuznetsov Jeffim N. , Durante Michael A. , Harbour J. William
Published: July 2021

Uveal melanoma (UM) is the most common primary eye cancer and has a high rate of metastasis with liver tropism. UM can be stratified according to metastatic risk into low risk Class 1 and high risk Class 2 tumor. Biallelic mutational inactivation of the tumor suppressor BAP1 is the quintessential molecular feature of Class 2 UM, but the mechanism by which loss of BAP1 leads to metastasis is poorly understood. We recently showed using single-cell RNA sequencing that the tumor microenvironment of primary and metastatic Class 2 UM is characterized by large numbers of inhibitory T cells and macrophages. Taken together, these findings led us to hypothesize that loss of BAP1 triggers changes in tumor-immune cell interactions that lead to an immunosuppressive microenvironment. To test this hypothesis, we performed RNA-seq and proteomic mass spectrometry in uveal melanocytes and UM cells engineered to lack BAP1 expression. These experiments identified a small cassette of immune modulatory genes that are up-regulated at the mRNA and protein levels following BAP1 loss. These proteins are known to drive monocytes towards type 2 macrophage polarization, which secrete cytokines that can inhibit T cell activation. Our findings suggest that altered interaction between UM cells and monocytes may be key to the emergence of metastatic competence in UM, and we are exploring this possibility with function experiments that may open the door to targeted therapy to subvert this process.

Citation Format: Christopher J. Kaler, James J. Dollar, Stefan Kurtenbach, Jeffim N. Kuznetsov, Michael A. Durante, J. William Harbour. Mechanisms of genomic-microenvironmental interactions in uveal melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2764.
Abstract 2770: Evaluation of tumor growth and baseline leukocyte infiltration in Charles River C57BL/6 and Balb/c mice engrafted with MC38, B16F10, RENCA and EMT6 cells
Authors: Eberle Christoph S. , Rowe Jenny , Fiore Ann , Mihalek Robert , Festin Stephen
Published: July 2021

Background: In vivo syngeneic models continue to be an important tool in cancer biology research. Charles River C57BL/6 and Balb/c mice, two frequently used inbred strains, can be implanted with different murine tumor cell lines. One aspect of syngeneic model development is to characterize the extent and composition of infiltrating leukocytes (TIL) of tumor-bearing mice. The ability to analyze TILs is a relevant factor in preclinical model selection for studying immunomodulatory responses.

Study Details: We evaluated growth rates and basal infiltrates in syngeneic models of solid mouse tumors. In four separate studies 7-week-old Charles River mice were implanted subcutaneously on the flank with either MC38 or B16F10 cells (C57BL/6) or with either RENCA or EMT6 cells (Balb/c), respectively. Mice were treated with isotype control IgG1 antibody (BioXcell clone MOPC-21) when tumors reached sizes of ~50-100 mm3. Surviving mice were euthanized when tumor burden reached or exceeded 2000 mm3 in line with internal IACUC guidelines. Spleen and tumors were then processed into single cell suspensions for immunophenotyping by multicolor acoustic-assisted flow cytometry.

Results: Growth kinetics of all four tumor cell lines, MC38, B16F10, RENCA and EMT6, were monitored throughout each study. At study termination murine leukocyte infiltration was found in all models. These TILs were immune profiled to determine the frequencies and composition of murine immune cells in the lymphoid and myeloid compartments. Phenotyping data confirmed the presence of viable CD45+ cells with total T-cells (CD3+), T-cell subsets (CD4+, CD8+ and Treg) and myeloid populations (CD11b+) including M1 and M2 TAMs as well as CD11c+ cells. M1/M2 TAM, CD8/Treg and CD4/CD8 ratios were also calculated, and tumor models were assessed based on their baseline TIL composition.

Conclusions: Four different allograft models of solid mouse tumors could be established using Charles River C57BL/6 and Balb/c mice. Their basal immune microenvironments could be characterized and therefore can be useful as pharmacodynamic readouts in efficacy studies of potential anti-cancer therapeutics.

Citation Format: Christoph S. Eberle, Jenny Rowe, Ann Fiore, Robert Mihalek, Stephen Festin. Evaluation of tumor growth and baseline leukocyte infiltration in Charles River C57BL/6 and Balb/c mice engrafted with MC38, B16F10, RENCA and EMT6 cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2770.
Abstract 278: Artificial intelligence accelerate drug discovery
Authors: Xie Weidong , Cheng Xing , Ding Zhengfang , Deng Riqiang , Gu Dawei
Published: July 2021

Drug discovery is resource intensive, and involves typical timelines of 10-20 years and costs that range from US$0.5 billion to US$2.6 billion. Artificial intelligence (AI) has recently started to gear-up its application in various sectors of the society and the pharmaceutical industry as a frontrunner beneficiary.Artificial intelligence can accelerate drug discovery and reduce costs by facilitating the rapid screening and identification of compounds. We have developed DM-AI drug discovery platform, including convolutional neural networks, decision treealgorithm, reinforcement learning, generative adversarial networks, big data, and knowledge graphs, along with structure and ligand-based high-throughput virtual screening , for new drug discovery and development. DM-AI optimizes biological activity,toxicity,physicochemical property. We used DM-AI to discover potent inhibitors of SHP2, PIM1, DNA-PK, kinases target implicated in solid tumor and other diseases.We started to train a biological activity prediction model on a database of the given target kinase inhibitors (positive set) and non-kinase targets molecules (negative set), and then predicted the activity of existing million data sets, obtained an initial output of thousands structures. We then evaluated these structures using a pharmacophore reward model on the basis of virtual chemical spaces of kinase inhibitors in complex with target protein. To narrow our focus to a smaller set of molecules for analysis, compounds with higher score were filtered to remove patents and applications molecules, also remove molecules bearing structural alerts and reactive groups.By day 7 after target selection, We had selected dozens structures with structural diversity for experimental validation. and by day 28, they were tested for in vitro inhibitory activity in an enzymatic kinase assay, active compounds accounted for up to 65% in some target models. This illustrates the utility of our DM-AI drug discovery platform for the successful, rapid discovery of drug candidates.

Citation Format: Weidong Xie, Xing Cheng, Zhengfang Ding, Riqiang Deng, Dawei Gu. Artificial intelligence accelerate drug discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 278.
Abstract 2785: Efficacy evaluation of candidate therapeutics in immuno-oncology by in vitro suppressive bioassays using macrophages
Authors: Janss Thibaut J. , Arnold Joahn , Lamy Juliette , Boelen Ellen
Published: July 2021

The increasing interest in the tumor microenvironment leads to a focus on new bioassays to represent all the players of the cancer immune response. Some of these players like Tumor Associated Macrophages (TAMs) often exhibit an immunosuppressive phenotype by downregulating the anti-tumor response. Their regulation mechanisms constitute an important target for new therapeutics. In order to study these mechanisms in a human model, suppressive bioassays mimicking the suppressive action of these cells on T cell activation, were developed. An important player in the tumor microenvironment are macrophages which possess important active and regulatory functions in both innate and adaptive immune responses. Classical activated macrophages, classified as M1-like macrophages, comprise immune effector cells with an acute inflammatory phenotype while the alternatively activated M2-like macrophages have suppressive and healing capacities. TAMs are present at high densities in solid tumors and share many characteristics with M2-like macrophages. In vitro assays using M1- and M2-like macrophages can be a first step to screen the effect of the test molecules on the phenotype and function of macrophages. For example, macrophage precursors display extraordinary plasticity in response to exogenous and endogenous stimuli which can result in a M2- or M1-like status. Using in vitro polarization and functional assays, the potential effect of molecules on M1- and M2-like macrophage generation and polarization can be screened. Next to that, the test molecules' impact on the functionality of macrophages can be evaluated in a suppressive assay. Here, the ability of the molecules to reverse the stimulating versus suppressive effect of the M1- and M2-like macrophages on T cells can be determined by measuring proliferation and cytokine production. The use of bioassays contributes to a better understanding of the tumor microenvironment. The steps needed to generate an anti-tumor response by the immune system will help to assess the functional potential of new drugs, design clinical trials and ultimately discover relevant biomarkers

Citation Format: Thibaut J. Janss, Joahn Arnold, Juliette Lamy, Ellen Boelen. Efficacy evaluation of candidate therapeutics in immuno-oncology by in vitro suppressive bioassays using macrophages [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2785.
Abstract 2786: A FcgR IIB tumor enriching anti-CD137 novel antibody to avoid liver Immuno-toxicity
Authors: Xue Jie , Chang Jen-Kuan , Dong Chen , Mao Yi-Ting , Raghavendra Pallavi , Li Ou , Issafras Hassan , Xu Wenfeng , Jiang Weidong
Published: July 2021

CD137 (4-1BB/TNFRSF9) is a costimulatory T-cell receptor belonging to the TNF receptor superfamily which is transiently expressed on T cell and NK cells. Targeting CD137 or its natural ligand 4-1BB ligand (4-1BBL) has important implications in many clinical conditions, including cancer. Anti-CD137 agonistic anti-CD137 antibodies have shown potent, often curative anti-tumor activity in preclinical models. In depth analysis revealed that 4-1BB-mediated anti-cancer effects are based on its ability to induce activation of cytotoxic T lymphocytes and high amounts of cytokines, including IFN-γ. However, this approach has suffered significant setback in the clinic due to either dose-dependent severe liver toxicity (e.g. Urelumab), or lack of single agent activity in the case of Utomilumab. Multiple therapeutics that attempt to restrict 4-1BB agonism to the tumor microenvironment and minimize systemic exposure have emerged. Here we present a novel antibody targeting 4-1BB receptor, named HLX25 with engineered Fc region to enhance the binding to FcγR IIB. In vitro, HLX25 induces comparable or superior CD137 activation and IL-2 release compared to Urelumab and Utomilumab, suggesting potentially a better safety profile. We showed that HLX25 binds to unique non-ligand competing epitope, which is different from both Urelumab and Utomilumab. In vivo, HLX25 shows potent dose-dependent single agent anti-tumor activity as low as 0.3mg/kg in MC38 model without sign of liver toxicity up to 20 mg/kg, suggesting superior efficacy and better safety profile.

Citation Format: Jie Xue, Jen-Kuan Chang, Chen Dong, Yi-Ting Mao, Pallavi Raghavendra, Ou Li, Hassan Issafras, Wenfeng Xu, Weidong Jiang. A FcgR IIB tumor enriching anti-CD137 novel antibody to avoid liver Immuno-toxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2786.
Abstract 2788: Deep characterization of tumor infiltrating leukocytes using a combination of flow cytometry and single-cell multiomics
Authors: Shi Xiaoshan , Nakamoto Margaret , Middlebrook Aaron , Huang Wei , Lo Evelyn , Neuhoefer Patrick T. , Bornheimer Scott , Ghanekar Smita
Published: July 2021

Cancer immunotherapies have the potential to induce durable anti-tumor immune responses; however the efficacy of these treatments varies among patients and partially depends on the complex interplay between malignant cells and non-malignant cells within the tumor microenvironment. In the tumor microenvironment, T cells play a crucial role in eliminating tumor cells and orchestrating anti-tumor immune responses; in contrast immunosuppressive cell types such as myeloid-derived suppressor cells can support tumor progression. Here we harnessed recent advances in single-cell multiomics that allow high-resolution functional characterization of intratumoral immune cells to improve our understanding of their dynamic relationships. Specifically, we utilized a One Solution Workflow to dissociate a solid tumor followed by in-depth dissection of the immune composition of the tumor microenvironment using both flow cytometry and the BD Rhapsody™ Single-Cell Analysis System. We characterized the cellular composition of tumor microenvironment using flow cytometry analysis. Importantly, by examining the expression of mRNA and surface protein markers at single-cell resolution, we unraveled the complexities of the tumor microenvironment, elucidated cellular heterogeneity and identified cell-type specific gene signatures. We propose this One Solution Workflow can be adopted to understand the transcriptional and proteomic phenotypes of tumor infiltrating leukocytes for development of novel immune therapeutic strategies for cancer.

For Research Use Only. Not for use in diagnostic or therapeutic procedures. Class 1 Laser Product.BD-23279 (v1.0) 1120BD, the BD Logo and Rhapsody are trademarks of Becton, Dickinson and Company or its affiliates. © 2020 BD. All rights reserved.

Citation Format: Xiaoshan Shi, Margaret Nakamoto, Aaron Middlebrook, Wei Huang, Evelyn Lo, Patrick T. Neuhoefer, Scott Bornheimer, Smita Ghanekar. Deep characterization of tumor infiltrating leukocytes using a combination of flow cytometry and single-cell multiomics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2788.
Abstract 2794: Blocking histamine receptor H1 on macrophages with anti-allergy drugs restores T-cell anti-tumor immunity
Authors: Li Hongzhong , Xiao Yi , Yao Jun , Yuan Xiangliang , Yin Xuedong , Zhang Yuan , Saito Yohei , Yagita Hideo , Gibbons Don , Yu Dihua
Published: July 2021

CD8+ T lymphocytes in tumors often become dysfunctional, prompting cancer immune escape and immunotherapy resistance. Restoring T-cell anti-tumor immunity remains an unmet challenge. Here, we show that histamine receptor H1 (HRH1) is highly expressed and tightly associated with T-cell dysfunction in human cancers. Histamine, the HRH1 ligand, is also increased in human and mouse tumors. HRH1 is predominantly expressed on tumor-associated macrophages (TAMs) and is activated by cancer cell-secreted histamine. The histamine/HRH1 axis in TAMs confers CD8+ T-cell dysfunction via increasing membrane localization of V-domain Ig suppressor of T cell activation (VISTA), an inhibitory immune checkpoint that promotes immunotherapy resistance. Blocking HRH1, genetically, or by anti-allergy drugs, reversed immunosuppressive TAMs, activated T-cells, and inhibited tumor growth in vivo. HRH1-targeting anti-allergy drugs more efficaciously enhanced immunotherapy response than anti-VISTA antibodies. Markedly, allergy-induced histamine also activates HRH1 on macrophages to induce T-cell dysfunction and immunotherapy resistance, which were reversed by the over-the-counter anti-allergy drug. Taken together, tumor cell-derived and allergy-released histamines activate HRH1 on TAMs to concede immunosuppression, pro-tumor function, and immunotherapy resistance, which can be negated by inexpensive and safer anti-allergy drugs.

Citation Format: Hongzhong Li, Yi Xiao, Jun Yao, Xiangliang Yuan, Xuedong Yin, Yuan Zhang, Yohei Saito, Hideo Yagita, Don Gibbons, Dihua Yu. Blocking histamine receptor H1 on macrophages with anti-allergy drugs restores T-cell anti-tumor immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2794.
Abstract 280: Sub-30-nm capsules for drug delivery
Authors: Ma Xiaowei , Zhang Ping , Cui Chao , Ling Chang-Chun , Cui Lina
Published: July 2021

Systemic treatment of cancer using long-circulating nanomedicines is promising due to their passive tumor targeting ability to achieve higher and more selective accumulation in tumors with irregular vascularization, a phenomenon known as extended permeation and retention (EPR) effect.1,2 Clinical use of nanometer-sized carriers, such as Doxil and Abraxane, to deliver chemotherapeutics to solid tumors is proven effective in highly vascularized tumors such as breast cancer, ovarian cancer, multiple myeloma, and Kaposi's sarcoma.3-5 Most nanomedicines that are being developed or approved so far have a diameter of around 100-200 nm for prolonged retention in highly angiogenic and densely vascularized tumors,6 however, they suffer from limited accumulation and poor penetration to the inner core of avascular or hypovascular tumors (such as prostate and pancreatic cancer),7-9 therefore nanomedicines small than 100 nm are more preferred for improved tumor penetration.10,11 Here we present our strategy to form cyclodextrin-based sub-30-nm nanocarriers, which allows easy drug encapsulation, and successful delivery of therapeutics to human tumor xenografts with significantly reduced tumor growth rates and improved survival rates.

Citation Format: Xiaowei Ma, Ping Zhang, Chao Cui, Chang-Chun Ling, Lina Cui. Sub-30-nm capsules for drug delivery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 280.
Abstract 2824: SHP2-PDHA metabolic axis is critical for adipocyte function by mitigating ER stress-induced adipocytes dysfunction: Implication for obesity-driven pancreatic ductal adenocarcinoma progression
Authors: Olou Appolinaire A. , Jack Jarrid , Manley Sharon , Ambrose Joseph , Walsh McKinnon , Eades Austin , VanSaun Michael N.
Published: July 2021

Obesity remains a major risk factor for the development of pancreatic cancer. Animal studies demonstrated that obese mice develop larger and more metastatic pancreatic cancer compared to their lean counterpart. Within the tumor environment are adipocytes that secrete adipokines, which promote inflammation and directly stimulate pancreatic cancer cells. Understanding the mechanisms that alter adipocyte function and adipokine secretion is critical for circumventing adipocyte-mediated influence on pancreatic cancer progression. Targeting of the Src homology region 2 containing protein tyrosine phosphatase (SHP-2) in the cancer cells has recently shown promising potential, yet the effect of SHP2 inhibition on stromal components has not been addressed. Here we demonstrate that SHP2 and the mitochondrial protein pyruvate dehydrogenase (PDHA) are critical mediators of adipocyte function, through mitigation of ER stress in adipocytes. In order to understand the role of SHP2 and PDHA in adipocytes, we performed Oil Red O staining and western blot assays on cultured adipocytes. We found that the transition from pre-adipocytes to fully developed adipocytes is marked by significant accumulation of phospho-SHP2, PDHA and the ER stress marker Bip. Chemical inhibition of SHP2 or PDHA augmented ER stress and impaired lipid production in the adipocytes; these effects were reversed by the ER stress inhibitor, 4-Phenylbyturic acid (4-PBA). Moreover, pancreatic cancer cells exposed to conditioned media from differentiated adipocytes, in which either SHP2 or PDHA was inhibited, are less aggressive and invasive compared to PDAC cells grown in conditioned media from control adipocytes with no inhibitor treatment. Therefore, our studies indicate that SHP2 and PDHA, in adipocytes, are key mediators of obesity-driven pancreatic cancer progression. Future studies will focus on understanding the SHP2-PDHA axis in adipocytes and further to elucidate how cancer cell extrinsic effects of SHP2 inhibition could alter PDAC progression.

Citation Format: Appolinaire A. Olou, Jarrid Jack, Sharon Manley, Joseph Ambrose, McKinnon Walsh, Austin Eades, Michael N. VanSaun. SHP2-PDHA metabolic axis is critical for adipocyte function by mitigating ER stress-induced adipocytes dysfunction: Implication for obesity-driven pancreatic ductal adenocarcinoma progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2824.
Abstract 2825: Mechanosurveillance by immunity targets mechanical compliance of metastatic cells
Authors: Tello-Lafoz Maria , Srpan Katja , Hu Jing , Romin Yevgeniy , Calo Annalisa , Hsu Katharine C. , Massagué Joan , Huse Morgan , Er Ekrem Emrah
Published: July 2021

Metastatic dissemination of cancer cells to vital secondary organs pose a lifelong lethal threat unless they can successfully be eliminated by the immune system. Interestingly, despite their ability to evade immunity at primary organs, the vast majority of disseminated cancer cells are killed at secondary organs, suggesting that the process of dissemination renders cancer cells sensitive to immunity. Understanding how dissemination renders cancer cells sensitive to immunity would therefore provide exciting new opportunities to target metastases, but the relationship between dissemination and the immune response is not clear. In our effort to study this relationship, we manipulated cancer cells' expression of myocardin related transcription factors A and B (MRTFA/B), which are central in regulation of actin cytoskeleton and are indispensable for metastatic invasion, dissemination and colonization. Surprisingly, despite MRTFA/B's established roles in promoting metastasis, we found that MRTFA/B expression in cancer cells downregulated metastatic colonization in fully immuno-competent mouse models. MRTFA/B mediated decrease in colonization was completely reversed upon immune cell depletion suggesting that the immune cells exploit MRTFA/B expression in cancer cells for cytotoxicity. Among various cancer cell parameters involved in immune cell activation, T- and NK cell activation most prominently correlated with MRTFA/B driven mechanical compliance of cancer cells. Mechanical compliance, also known as stiffness, of antigen presenting cells and synthetic substrates are well-known to activate T- and NK cells through biophysical and mechano-chemical signal transduction. As such, perturbing cancer cells' stiffness downregulated the immune response and cytotoxicity. Thus, we termed this process mechanosurveillance. Our work on mechanosurveillance will help us understand how disseminated cancer cells are targeted by immunity and how they evade it at secondary organs. Importantly the concept of mechanosurveillance will help guide the use of existing therapeutics that alter tissue stiffness and immune boosting therapies to combat metastases.

Citation Format: Maria Tello-Lafoz, Katja Srpan, Jing Hu, Yevgeniy Romin, Annalisa Calo, Katharine C. Hsu, Joan Massagué, Morgan Huse, Ekrem Emrah Er. Mechanosurveillance by immunity targets mechanical compliance of metastatic cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2825.
Abstract 2832: Targeted genomic landscape of metastases compared to primary tumors in non-small-cell lung cancer
Authors: Chen Bing , Dong Gaochao , Liang Yingkuan , Zhang Te , Zhang Junling , Chen Shiqing , Jiang Feng , Xu Lin
Published: July 2021

The genomic landscape of primary non-small-cell lung cancer (NSCLC) has been well described. However, little is known about cohort genomic alterations (GA) landscape in NSCLC metastases, and how it compares with primary tumors in general. The genomic landscape of metastases may be important for clinical treatment and prognosis. In this retrospective study, 37 cases of surgically resected NSCLC were studied, of which 19 were non-metastatic NSCLC and 18 were metastatic NSCLC. Sixty formalin-fixed paraffin-embedded (FFPE) tissue samples from 37 NSCLC patients, including the primary lesion of non-metastatic NSCLC (Group A, N =19), the primary lesion of metastatic NSCLC (Group B, N =18), and lymph node lesions (Group C, N =23), were included to detect somatic mutations using a 1021-gene panel by next-generation sequencing (NGS). In this study, twenty-nine patients (78.4%) were male. Nineteen patients with lung adenocarcinoma (LUAD) and 18 with lung squamous cell carcinoma (LUSC) were included. In addition, there were significant differences between Groups A and B in stage T and stage N (P < 0.0001). Overall, EGFR (33.3% vs 15.8%) and TERT (16.7% vs 5.2%) were numerically more frequently mutated in metastases, but differences were not statistically significant. Moreover, we found that ARID1A may be closely related to metastasis in LUSC (P = 0.029). These data support the theory that primary and metastatic NSCLC encompass a uniform distribution of common genomic alterations tested by next-generation sequencing targeted panels. While ARID1A may be closely related to metastasis in LUSC.

Citation Format: Bing Chen, Gaochao Dong, Yingkuan Liang, Te Zhang, Junling Zhang, Shiqing Chen, Feng Jiang, Lin Xu. Targeted genomic landscape of metastases compared to primary tumors in non-small-cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2832.
Abstract 2842: 6-gingerol attenuates metastasis in aged mice model of colorectal cancer through modulation of epithelial-mesenchymal transition mediators genes and e-cadherin upregulation
Authors: Ajayi Babajide Oluwaseun , Adenigba Ayomiposi , Adeshina Anuoluwapo
Published: July 2021

Colorectal cancer (CRC) is the third most common cancer in America, and the third cause of cancer related death. Metastasis is the major cause of death in patient with CRC. Many CRC are likely to spread from primary site to the other parts of the body before and after detection of malignant tumor. 6-gingerol, the bioactive component of fresh ginger have been reported to have myriad of biological activities including anti-inflammatory and antioxidant properties. We have previously reported the anti-CRC effect of 6-gingerol. However there is dearth of information on the anti-metastatic effect 6-gingerol in CRC. Thus, the effect of 6-gingerol on epithelial-mesenchymal transition mediators' genes and E-Cadherin repression in aged mice model of CRC was assessed in this study. Following institutional ethical approval. Aged male BALB/c mice (10 months) were divided into four groups of 10 mice each. Mice were fed with AIN-93 diet and allowed access to water ad libitum. Group 1 mice serve as control, group 2 mice received 6-gingerol 10mg/kg/day by oral gavage for 10 weeks. Group 3 mice received a single dose of azoxymethane (AOM) 10mg/kg (iP) and on the seventh day they received three cycles of dextran sulfate sodium (DSS) 2% (W/V) in drinking water ( each cycle is define as one week of DSS followed by two weeks of plain drinking water) Group 4 mice received 6-gingerol 10mg/kg/day by oral gavage in combination with single dose of azoxymethane 10mg/kg (iP) and on the seventh day they received three cycles of dextran sulfate sodium salt 2% (W/V) in drinking water. Mice were humanely sacrificed under anesthesia and colon tissues were processed for RT-PCR and multiplex immunohistochemistry analysis. Mice that received both 6-gingerol and AOM and DSS showed a significant decreased expression of Lymphocyte function-associated antigen-1, N-cadherin, SNAIL 1, ZEB 1, AP4 and HMGA1 with concomitant Upregulation miR-34, miR-15a, miR-218 and miR-302 expression as compared with the AOM and DSS only group. The result obtained from this study showed 6-gingerol inhibited metastasis through modulation of epithelial-mesenchymal transition mediators' genes and E-Cadherin upregulation, and as such 6-gingerol could be a promising anti-metastatic agent in CRC treatment.

Citation Format: Babajide Oluwaseun Ajayi, Ayomiposi Adenigba, Anuoluwapo Adeshina. 6-gingerol attenuates metastasis in aged mice model of colorectal cancer through modulation of epithelial-mesenchymal transition mediators genes and e-cadherin upregulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2842.
Abstract 2847: Human CYP1B1 induces cancer cell metastasis through Sp1-mediated activation of uPA-uPAR signaling pathway
Authors: Kwon Yeo-Jung , Shin Sangyun , Chun Young-Jin
Published: July 2021

Human CYP1B1 is known as a major metabolizer for estrogen and shows tumor-specific hyper-expression. To explore the role of CYP1B1 on metastasis of human cancer cells, we studied the effects of CYP1B1 inhibition and activation in MCF-10A, MCF-7, MDA-MB-231, and HeLa cells. CYP1B1 significantly induced expression of urokinase-type plasminogen activator receptor (uPAR) as well as uPA. Treatment with DMBA, a well-known CYP1B1 inducer, enhanced uPAR expression while CYP1B1 siRNA and TMS (tetramethoxystilbene), a specific CYP1B1 inhibitor, suppressed uPAR. We also found that CYP1B1 activated uPAR signaling through regulation of related factors including integrin β1 and α5. Interestingly, uPAR overexpression only caused a significant increase of integrin β1 and α5 in protein levels, indicating protein degradation may play an important role in regulating integrin protein level. Surprisingly, CYP1B1 down-regulated p53 expression through MDM2 activation and nutlin-3a, an inhibitor for MDM2, blocked the promoting effects of DMBA on uPAR, which explains that CYP1B1 induced uPAR expression through regulation of MDM2-p53 system. In addition, we revealed that CYP1B1 regulates uPAR through activation of Sp1 transcription factor. CYP1B1 and its enzymatic product, 4-OHE2, promote Sp1 expression levels in both mRNA and protein levels and subsequently Sp1 elevates the transcriptional activity of uPAR through binding to promoter region of uPAR. Taken together, our data suggest that CYP1B1 promotes cancer cell metastasis via activating uPAR pathway which is one of the targets of p53 and Sp1 signaling.

Citation Format: Yeo-Jung Kwon, Sangyun Shin, Young-Jin Chun. Human CYP1B1 induces cancer cell metastasis through Sp1-mediated activation of uPA-uPAR signaling pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2847.
Abstract 2850: Astrocyte-breast cancer interactions facilitate brain metastasis
Authors: Dangoor Sahar Israeli , Hadad Ori , Satchi-Fainaro Ronit
Published: July 2021

Diagnosis and treatment of breast cancer patients have been considerably improved, including those at the metastatic stage. However, brain metastasis still presents a major clinical challenge, and it is a leading cause of death. Unfortunately, there are no targeted therapies specific for this secondary tumor, and the current treatment modalities present limited clinical outcomes. Thus, better understanding of the pathways involved in the metastatic cascade, which lead to brain metastases colonization and progression, is essential. To that end, we investigated the mutual effects of astrocytes and breast cancer cells, using the traditional 2-dimensional (2D) cell-culture and a newly-established 3-dimensional (3D) model of multicellular breast cancer brain metastases spheroids. We found that astrocytes enhance the proliferation, migration and invasion of breast cancer cells, suggesting a supportive role for astrocytes in the metastatic process of breast cancer in the brain. Using a protein array, we identified several cytokines which were elevated following the co-culturing of breast cancer cells with astrocytes, suggesting them as potential key candidates involved in breast cancer-brain stroma interactions. One of these cytokines was MCP-1, which is a chemokine that is associated with poor prognosis in breast cancer patients. When it was functionally neutralized, breast cancer cell proliferation and migration rates were decreased, in addition to reduced invasion shown in our 3D-spheroid model. Considering these results, MCP-1 is proposed as one of the key factors promoting breast cancer brain metastasis. Although further validation using in vivo models is still required, this study deepens our understanding of the contribution of the metastatic breast cancer cells-brain microenvironment crosstalk to brain colonization. Taken together, our findings may be utilized to provide a novel therapeutic approach for the prevention of breast cancer brain metastasis following resection of the primary lesion.

Keywords: Breast Cancer, Brain Metastasis, Brain Microenvironment, Astrocytes, Cytokines, Chemokines, MCP-1, CCL2, 3D Tumor Spheroids.

Citation Format: Sahar Israeli Dangoor, Ori Hadad, Ronit Satchi-Fainaro. Astrocyte-breast cancer interactions facilitate brain metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2850.
Abstract 2853: Transposon-activated POU5F1B promotes colorectal cancer growth and metastasis
Authors: Simó-Riudalbas Laia , Planet Evarist , Offner Sandra , Duc Julien , Abrami Laurence , Dind Sagane , Coudray Alexandre , Coto-Llerena Mairene , Piscuoglio Salvatore , Andersen Claus L. , Bramsen Jesper Bertram , Trono Didier
Published: July 2021

The human genome contains some 4 million transposable element (TE)-derived sequences, which collectively influence gene expression from early embryogenesis to adulthood. The illegitimate production of oncogene-encoding TE-driven transpochimeric gene transcripts (TcGTs) has been noted in tumors, but their participation in the oncogenic process has seldom been demonstrated. Here we describe how the aberrant de-repression of a primate-specific LTR66 endogenous retroviral promoter generates a TcGT overexpressing the great-ape-restricted POU5F1B retrogene in ~65% of 286 colorectal cancers (CRC) patients, correlating with more advanced tumor stages and shorter relapse-free and overall survival. We further demonstrate that the POU5F1B protein stimulates the clonogenic and proliferation capacity of human CRC cell lines in vitro, and their tumorigenic and metastatic potential in mouse xenotransplantation models. Although POU5F1B is a retrotransposition-mediated derivative of the POU5F1/OCT4 transcription factor, its product is a predominantly cytoplasmic protein enriched in membranes that associates with mediators of signal transduction, notably the ERBB2 receptor tyrosine kinase and several of its known interactors. POU5F1B overexpression results in activating genes involved in signaling, and conditioned medium of POU5F1B-overexpressing cells enhances the clonogenic potential of CRC cells in trans. As POU5F1B is encoded by an apparently non-essential gene only lowly expressed in normal tissues, and as POU5F1B-containing TcGTs are detected in other tumors besides CRC, it represents an attractive target for the development of cancer therapies.

Citation Format: Laia Simó-Riudalbas, Evarist Planet, Sandra Offner, Julien Duc, Laurence Abrami, Sagane Dind, Alexandre Coudray, Mairene Coto-Llerena, Salvatore Piscuoglio, Claus L. Andersen, Jesper Bertram Bramsen, Didier Trono. Transposon-activated POU5F1B promotes colorectal cancer growth and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2853.