bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2720: Perivascular accumulation of immunosuppressive cells in the stroma of human triple negative breast carcinomas: implications for immunotherapy
Authors: Moamin Mohammed Ridha , Allen Richard J. , Stavrou Nicholas , Nagy Mate , Juncker-Jensen Anna , Lewis Claire E. 
Published: July 2021

Abstract
A distinct subset of tumor-associated macrophages (TAMs) accumulate around blood vessels in mouse tumors where they stimulate tumor angiogenesis and various steps in the metastatic pathway. They also limit tumor responses to frontline anti-cancer therapies like irradiation and chemotherapy, and dampen anti-tumor immunity by recruiting regulatory T cells and suppressing the proliferation of T cells. However, the presence and phenotype of such perivascular (PV) TAMs have yet to be investigated in human tumors. In this study we have used MultiOmyx multiplex immunofluorescence coupled to advanced analytics to compare the distribution and phenotype of TAMs, CD4+ and CD8+ T cells, and CD4+FOXP3+ regulatory T cells (Tregs) in PV vs non-PV areas in the stroma and tumor cell islands (TCIs) of 40 human triple negative breast carcinomas (TNBCs) - and whether this was altered after neoadjuvant chemotherapy. Tregs along with distinct subsets of TAMs and CD4+ and CD8+ T cells were found to preferentially accumulate in PV areas (ie. within 50um of CD31+ blood vessels), especially in the tumor stroma. CD163+ TIM3+ TAMs often made direct contact with the abluminal surface of blood vessels in these sites, and at increased numbers in in chemotherapy-treated TNBCs. This suggests they may regulate the formation/function of blood vessels in relapsing tumors. In both untreated and chemotherapy-treated tumors, a major subset of CD163+ TAMs lacked PDL1 expression and also accumulated preferentially in stromal PV areas, where many made direct contact with PD1- CD4+ or PD1- CD8+ T cells as well as FOXP3+CD4+ Tregs. We are currently investigating whether such close contact of these subsets of T cells with two immunosuppressive cell types (ie. CD163+ TAMs and Tregs) inhibits their subsequent anti-tumor functions. Taken together, our study shows that, as T cells cross blood vessels into TNBCs, they encounter a high density of at least two immunosuppressive cell types in the perivascular niche. This could lead to the inactivation of T cells before they migrate further into tumors, thereby reducing anti-tumor immunity. It could also limit the efficacy of cancer immunotherapies mediated by activated T cells. Identifying and targeting the mechanism causing these suppressive cells to accumulate and interact in such sites could remove this inhibition.
Citation Format: Mohammed Ridha Moamin, Richard J. Allen, Nicholas Stavrou, Mate Nagy, Anna Juncker-Jensen, Claire E. Lewis. Perivascular accumulation of immunosuppressive cells in the stroma of human triple negative breast carcinomas: implications for immunotherapy [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 2720.
Abstract 2722: Reprogramming of tumor-associated macrophages to M1-like macrophages using M1 macrophage-derived exosomes engineered to target IL-4 receptor and enhance M1 polarization
Authors: Lee Byungheon , Gunassekaran Gowri Rangaswamy 
Published: July 2021

Abstract
Tumor-associated macrophages (TAMs) are M2-polarized protumoral macrophages, whereas inflammatory macrophages are M1-polarized antitumoral macrophages. Interleukin-4 receptor (IL-4R) is expressed in M2 macrophages at higher levels than M1 macrophages. In this study, we aimed to inhibit tumor growth using M1 macrophage-derived exosomes engineered to target IL-4R and reprogram TAMs into M1 macrophages. M1 exosomes were transfected with NF-κB p50 siRNA and miR-511-3p as cargoes to enhance M1 polarization (Exo-si/mi) and surface-modified with IL4RPep-1, an IL-4R-binding peptide, to target IL-4R of M2 macrophages (IL4R-Exo-si/mi). The engineered exosomes showed sizes similar with parental exosomes, and IL4RPep-1 was retained on the exosomes and not degraded in the presence of serum up to 24 h. IL-4R-targeted exosomes were internalized into M2 macrophages and delivered cargoes through IL-4R more efficiently than untargeted exosomes. Whole-body fluorescence imaging showed that IL-4R-targeted exosomes homed to tumors at higher levels than to liver unlike to untargeted exosomes. Intravenous administration of IL4R-Exo-si/mi decreased the levels of M2 markers and immune-suppressive cells including TAMs, while increasing the levels of M1 markers and immune-stimulatory cells including M1 macrophages in tumor and spleen and subsequently inhibited tumor growth more efficiently than Exo-si/mi as well as M1 exosomes. These results suggest that IL-R-targeted exosomes carrying NF-κB p50 siRNA and miR-511-3p inhibit tumor growth by reprogramming TAMs into M1 macrophages and increasing anti-tumor immunity and hold a potential for caner immunotherapy.
Citation Format: Byungheon Lee, Gowri Rangaswamy Gunassekaran. Reprogramming of tumor-associated macrophages to M1-like macrophages using M1 macrophage-derived exosomes engineered to target IL-4 receptor and enhance M1 polarization [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 2722.
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

Abstract
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

Abstract
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&lt;0.01), CD8+granzymeB+CD45RO+, (p&lt;0.01) and CD8+granzymeB+CD44+CD45RO+ cells (p&lt;0.01) compared to primary tumors. Both stromal and intratumoral CD20+ cells were higher in recurrent tumors (p=0.03, p&lt;0.01). Stromal CD73+Coll-I+ cells were lower in recurrent tumors (p&lt;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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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.