bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 634: The impact of IL-4 on migration in triple negative breast cancer cell lines
Authors: Andrews Portia , Kimbro Kevin S.
Published: June 2023

In the United States, triple-negative breast cancer (TNBC), an aggressive form of breast cancer (BCa), disproportionately affects women of African descent and contributes to racial disparities in BCa mortality. Various risk factors are thought to drive this disparity, including socioeconomic status, tumor characteristics, comorbidities, and quality of treatment. Complex diseases and environmental insults all invoke and/or disrupt host inflammation, with individual immune and variable responses to therapeutic agents being dictated in part by genetic variants. This latter has seldom been studied with regards to precision diagnosis and prognosis of individual patient responses, in the frame of geographical ancestry and TNBC. Interleukin-4 receptors (IL4R) have been shown to be upregulated in many epithelial cancers and could be a promising target for metastatic tumor therapy. Yet, IL4R overexpression and/or IL4R variants have not been evaluated in TNBC cell lines of varying genetic ancestry. In view of this, we investigated the effects of IL-4 treatment in four TNBC cell lines varying in proportions of West African Ancestry (WAA) in each cell line: MDA-MB-231 (0.019), HCC-1143 (0.012), HCC-1806 (0.807), MDA-MB-468 (0.803). The frequency of IL4R variants also differed in each cell line. We used high content live cell imaging to evaluate the effect of IL-4 treatment on cellular migration by measuring wound healing. We show that IL-4 treatment on TNBC cells enhances migration as indicated by increased wound healing when compared to non-treated cells. High glucose, at a concentration of 10 g/L, delays cell migration in IL4-treated TNBC cells. This data suggests that comorbidities, such as high glucose levels (i.e., uncontrolled diabetes) could impact tumor behavior and mortality disparities between populations.

Citation Format: Portia Andrews, Kevin S. Kimbro. The impact of IL-4 on migration in triple negative breast cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 634.
Abstract 6353: Combination of poly-γ-glutamic acid with anti PD-L1 blockade leads to improved anti-tumor activity in murine models
Authors: Jang Jae-Pyung , Choi Jae Chul , Hahm Kyung-Soo , Lee Do Young , Park Young-Chul
Published: June 2023

Immune checkpoint inhibitors(ICIs) have been shown to be key role in cancer therapy. Especially, antibodies targeting programmed cell death ligand1(PD-L1) promoted T cells to activate immune response against cancer cells. However, the current ICIs still have insignificant to motivate effective antitumor immunity by blocking PD-1/PD-L1 pathway and the regulatory network of PD-L1 by cytokines in the tumor microenvironment remains to be uncovered. Previously, we have reported the anti-cancer effects of combination treatment with a natural product, poly-γ-glutamic acid(γ-PGA), and PD-1 blockade. Now, we showed that combination therapy with anti PD-L1 and γ-PGA overcome the limits of ICIs and enhance anticancer effect in murine melanoma(B16F10). In vivo efficacy studies were performed by intraperitoneal injection of antibody targeting PD-L1 and oral administration of γ-PGA in C57BL/6 model transplanted with B16F10 melanoma cell. Systemic changes in immunophenotype were measured by tumor volume size and survival rate. The splenocytes were isolated from mice and the populations of myeloid-driven suppressor cells(MDSCs) and CD4/CD8 T cells were analyzed by FACS. Combination therapy with γ-PGA and anti PD-L1 blockade showed dramatically more suppressive tumor growth and enhanced long-term survival rate than other monotherapic treatment. In addition, γ-PGA was shown to play a role in the immune modulation of tumor microenvironment by activation of T cells and inhibition of MDSCs so as to prevent tumors from immune escape. Therefore, we suggest that γ-PGA can promote ICI-mediated antitumor responses by the transformation of "cold tumors" into “hot tumors”

Citation Format: Jae-Pyung Jang, Jae Chul Choi, Kyung-Soo Hahm, Do Young Lee, Young-Chul Park. Combination of poly-γ-glutamic acid with anti PD-L1 blockade leads to improved anti-tumor activity in murine models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6353.
Abstract 6362: Overcoming resistance to immunotherapy due to loss of antigen presentation
Authors: Miller Brian C. , Choutri Yacine , Abosy Rose Al , Huang Amy , Cox Emily K. , Zimmerman Matthew P. , Chong Wan Lin , Vietor Katherine J. , Collier Jenna , Weiss Sarah A. , Sen Debattama , Haining W. Nicholas , Sharpe Arlene H.
Published: June 2023

Resistance to immune checkpoint inhibitors represents a major therapeutic challenge, as only 40% of patients with melanoma (and less with other tumor types) have a long-term response to anti-PD-1 therapy. Resistance can arise because of somatic mutations in cancer cells that allow evasion of T cell-mediated killing. One commonly acquired resistance mutation in melanoma, loss of beta-2 microglobulin (B2m), prevents T cell killing by hiding cancer cells from CD8+ T cell recognition. To understand the failed immune response against resistant tumors, we used single-cell RNA-seq to characterize tumor-infiltrating immune cells in antigen presentation-deficient human melanoma biopsies and CRISPR-modified mouse melanoma tumors. Our data demonstrate an increase in immunosuppressive M2-like macrophages and absence of CD8+ T cells in B2m-null tumors. To overcome this resistance, we treated tumor-bearing mice with CD40 agonist antibody, which promotes differentiation of macrophages towards a pro-inflammatory phenotype and increases dendritic cell priming of CD8+ T cells. Treatment with CD40 agonist reduced tumor growth and improved tumor clearance in B2m-null melanoma and colorectal cancer models. To determine how CD40 agonist treatment works, we depleted different immune populations from the tumor microenvironment. We hypothesized that by depleting M2 macrophages, CD40 agonist treatment would remove an immunosuppressive brake to allow natural killer (NK) cells to kill tumor cells lacking MHC expression. To our surprise, NK cells were not required for the efficacy of CD40 agonist. Instead CD8+ T cells were required, even though the CD8+ T cells cannot directly recognize the tumor cells. scRNA-seq identified a transcriptionally unique state of CD8+ T cells that is recruited to the tumor microenvironment after CD40 agonist treatment. These CD8+ T cells produce IFNγ, which is required for the efficacy of CD40 agonist treatment. These data demonstrate that CD8+ T cells, a key mediator of anti-tumor immunity, can still be recruited to control tumors deficient in antigen presentation. More broadly, they suggest that strategies to activate CD8+ T cells may be effective even in the context of acquired resistance to checkpoint inhibitor therapy.

Citation Format: Brian C. Miller, Yacine Choutri, Rose Al Abosy, Amy Huang, Emily K. Cox, Matthew P. Zimmerman, Wan Lin Chong, Katherine J. Vietor, Jenna Collier, Sarah A. Weiss, Debattama Sen, W. Nicholas Haining, Arlene H. Sharpe. Overcoming resistance to immunotherapy due to loss of antigen presentation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6362.
Abstract 6363: Short-term exposure of cancer cells to ultra-high concentrations nitric oxide induces PDL-1 upregulation
Authors: Epshtein Yana , Goldshtein Matan , Chaisson Selena , Monson Jedidiah , Johnson Matt , Avniel Amir , Lisi Steve , Confino Hila
Published: June 2023

Background: We have previously shown that treating mouse colon carcinoma (CT26) tumor-bearing mice with ultra-high concentrations of nitric oxide (UNO) upregulates innate and adaptive immune cells both locally and systemically. As immune-checkpoint molecules, such as the PD-L1 receptor PD-1, are expressed on immune cells, our group assessed the potential synergy of UNO and an anti-PD-1 antibody for treating tumors in vivo. Indeed, primary tumors regressed in 53% of the CT26 tumor-bearing mice treated with UNO and anti-PD-1. Moreover, these mice were primary and distant tumor-free and survived 100 days post-treatment. In the current study, we assessed PD-L1 expression levels on cancer cells in vitro following short-term exposure to UNO.

Methods: CT26 cells were exposed to 25,000 - 100,000 ppm NO for 10 - 60 seconds. The percentage of PD-L1-expressing cells was assessed by flow cytometry analysis.

Results: 1. Exposing CT26 cells to 100,000 ppm NO for 10 seconds upregulated PD-L1 expression. 85.1% of cells, which are still viable or at early apoptosis, expressed PD-L1 compared to 71% of untreated cells (p<0.0001). Exposure of CT26 cells to 25,000 ppm or 50,000 ppm NO for 10 seconds did not change the level of PD-L1 expression significantly. 2. Exposure to 100,000 ppm NO for 30 seconds further increased the percentage of cells expressing PD-L1 to 96.7% (p<0.0001 compared to untreated cells). For this duration, lower concentrations of NO also increased PD-L1-expression in viable early apoptotic CT26 cells to 82.8% - 92.3% when treated with 25,000 ppm or 50,000 ppm NO, respectively. 3. Finally, a 60 second exposure of CT26 cells to 25,000 ppm, 50,000 ppm, or 100,000 ppm NO resulted in the highest percentage of PD-L1 expressing cells. Approximately 95% of all viable or early apoptotic CT26 cells express PD-L1 under all NO conditions. This finding is significant compared to 71% of cells in the control arm that express PD-L1 (p<0.0001).

Conclusion: Short-term exposure (10 - 60 seconds) to UNO upregulates PD-L1 expression on cancer cells in an NO concentration-dependent and exposure-dependent process. A 10-second exposure to 100,000 ppm NO is sufficient to significantly induce PDL-1 expression. Therefore, NO therapy can potentially be combined with anti-PD-L1 antibody treatment to improve therapeutic outcomes.

Citation Format: Yana Epshtein, Matan Goldshtein, Selena Chaisson, Jedidiah Monson, Matt Johnson, Amir Avniel, Steve Lisi, Hila Confino. Short-term exposure of cancer cells to ultra-high concentrations nitric oxide induces PDL-1 upregulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6363.
Abstract 637: Cytokine CCL9 mediates oncogenic Kras-induced pancreatic acinar-to-ductal metaplasia through ROS
Authors: Liou Geou-Yarh , Messex Justin K. , Adams Kiyah L.
Published: June 2023

Pancreatic ductal adenocarcinoma (PDAC) can be originated from acinar-to-ductal metaplasia (ADM). Pancreatic acini harboring oncogenic Kras mutations are transdifferentiated to a duct-like phenotype that further progresses to become pancreatic intraepithelial neoplasia (PanIN) lesions, giving rise to PDAC. Although ADM formation is frequently observed in KrasG12D transgenic mouse models of PDAC, the exact mechanisms of how oncogenic KrasG12D regulates this process remain an enigma. Herein we reveal a new downstream target of oncogenic Kras, cytokine CCL9 during ADM formation. Exogenously treating primary pancreatic acini with recombinant CCL9 in a 3D organoid culture system drives ADM formation. Furthermore, knockdown of CCL9 in KrasG12D-expressed acini reduced KrasG12D-induced ADM. Higher levels of reactive oxygen species were detected in the KrasG12D-expressed pancreatic acini, and knockdown of CCL9 in these acini dramatically decreased ROS levels. Depletion of ROS in 3D organoid culture of CCL9-expressed acini reduced CCL9-induced ADM formation. In transgenic mice of p48cre:KrasG12D, depletion of CCL9 through its specific neutralizing antibody attenuated ADM formation as well as PanIN structures. Hence, our results provided insight into how oncogenic Kras regulates pancreatic ADM through a new downstream target molecule, CCL9.

Citation Format: Geou-Yarh Liou, Justin K. Messex, Kiyah L. Adams. Cytokine CCL9 mediates oncogenic Kras-induced pancreatic acinar-to-ductal metaplasia through ROS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 637.
Abstract 6370: Cell-based luminescent reporter bioassays for immunotherapies targeting macrophage effector functions
Authors: Mitchell Jonathan , Gilden Julia , Hartnett Jim , Stecha Pete , Krishnan Gopal B. , Fan Frank , Cong Mei , Grailer Jamison
Published: June 2023

Macrophages play a key role in the elimination of cancer via phagocytosis and presentation of tumor antigens to T cells. Antibody-dependent cellular phagocytosis (ADCP), mediated by macrophages and other myeloid cells, is an important mechanism-of-action (MoA) for antibody-based cancer immunotherapies. Therapeutic strategies that enhance ADCP and other macrophage effector functions can augment direct tumor destruction and enable anti-tumor immunity. ADCP is initiated by binding of antibody Fc domains to Fc gamma receptors (FcgRs), and FcgR crosslinking results in signal transduction that drives inflammatory cytokine production and phagocytosis. Simultaneously, macrophage effector functions are regulated by immune checkpoints. Blockade of these immune checkpoints (e.g., SIRPa, ILT4), or agonists of activation pathways (e.g., TLR receptors) can enhance macrophage effector functions. Current assays to measure macrophage function are tedious, low throughput, and highly variable. Here, we present a suite of cell-based luminescent reporter bioassays to measure macrophage effector functions: 1) A monocytic cell background was used to measure ADCP via endogenous expression of multiple FcgRs and integration of a FcgR-activation dependent luminescent reporter. 2) Direct detection of cellular phagocytosis can be measured using the HiBiT split luciferase system. 3) Modulation of macrophage-expressed immune checkpoints can be determined using cell-based reporter bioassays for SIRPa/CD47 and ILT4/HLA-G. 4) Activation of TLRs 1, 2, 4, 5 and 6 can be measured using a TLR reporter bioassay. These bioassays are robust, easy-to-use, and are pre-qualified according to ICH guidelines. Together, these novel reporter bioassays provide a robust, high-throughput toolbox to facilitate discovery and development of immunotherapies targeting macrophage effector functions.

Citation Format: Jonathan Mitchell, Julia Gilden, Jim Hartnett, Pete Stecha, Gopal B. Krishnan, Frank Fan, Mei Cong, Jamison Grailer. Cell-based luminescent reporter bioassays for immunotherapies targeting macrophage effector functions. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6370.
Abstract 6371: Identification of a lymphocyte activation gene-3-binding peptides using phage displayed-peptide libraries for cancer immunotherapy
Authors: Lee Seok-Min , Lee Byungheon
Published: June 2023

Treatment with immune checkpoint blockades against CTLA-4, PD-1, and PD-L1 is currently giving improved results to many cancer patients. Nevertheless, a significant proportion of cancer patients treated with immune checkpoint blockades are still largely unsatisfied with the benefits of these drugs. One of the reasons is that cancer cells also use other immune checkpoints such as lymphocyte activation gene-3 (LAG-3), T cell immunoreceptor with Ig and ITIM domains, and T-cell immunoglobulin and mucin-domain containing-3. In this study, we screened phage displayed-peptide libraries for peptides that selectively bind to LAG-3. After five rounds of screening, we selected a candidate peptide and named it LAG3Pep. LAG3Pep preferentially bound to human LAG-3-transfected HEK 293T cells over mock-transfected cells and phorbol myristate acetate/ionomycin/chloroquine-stimulated Jurkat T cells over unstimulated cells. LAG3Pep also bound to splenocytes isolated from tumor-bearing mice, suggesting the reactivity across human and mouse LAG-3. Pull-down assay of human recombinant LAG-3 protein using biotin-labeled LAG3Pep yielded a protein band of LAG-3. LAG3Pep showed a high-affinity binding to the LAG-3 protein in SPR assays. Pre-treatment of a LAG-3-blocking antibody competed the binding of LAG3Pep to stimulated Jurkat T cells. LAG3Pep inhibited the binding of the LAG-3 protein to THP-1 cells expressing HLA-DR, a well-known LAG-3 ligand. In addition, treatment of T-cells with LAG3Pep in combination with a PD-L1-blocking antibody recovered the secretion of IL-2 by T-cells which was suppressed by FGL-1, another ligand of LAG-3, secreted from HepG2 tumor cells in co-culture. Intravenous administration of LAG3Pep in combination with a PD-L1-blocking peptide showed an anti-tumor growth activity in the MC38 syngeneic mouse colon tumor model. These results show that LAG3Pep acts as LAG-3 blockade and, in combination with a PD-L1-blockade, could be a promising tool for cancer immunotherapy.

Citation Format: Seok-Min Lee, Byungheon Lee. Identification of a lymphocyte activation gene-3-binding peptides using phage displayed-peptide libraries for cancer immunotherapy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6371.
Abstract 6436: 1C3, a novel non-blocking anti-human TNFR2 antibody, promotes effector T cell responses and demonstrates potent anti-tumor activity
Authors: Yang Yongfei , Cao Shuzhen , Zhang Wenjiao , Zhang Jing , Shao Zhe , Jiang Xueyuan , Duan Qingya , Tian Maopeng , Mei Sen , Yu Zhaoxue
Published: June 2023

Regulatory T cells (Tregs) are a subset of immunosuppressive CD4+ T lymphocytes which can inhibit effector T cell activity in the tumor microenvironment (TME) and promote tumor growth. Depletion of Tregs is a promising immunotherapy strategy for treatment of cancers. Tumor necrosis factor receptor-2 (TNFR2) is highly expressed on both Tregs and many kinds of tumor cells, making it a potential candidate for tumor-targeted antibody therapy. A fully human, non-blocking agonist antibody to TNFR2, 1C3, was generated from RenMab™ mice, which contain the full human immunoglobulin variable domain. 1C3 was selected from a large panel of antibody candidates after unbiased, high-throughput efficacy screening conducted in syngeneic tumor mouse models established in humanized TNFR2 mice. In vitro, 1C3 effectively promoted proliferation of cytotoxic T cells. Subsequently, the in vivo anti-tumor efficacy and safety of 1C3 was evaluated in syngeneic tumor mouse models expressing human TNFR2. The results demonstrated that 1C3 monotherapy significantly inhibited tumor growth in a dose-dependent manner. In combination with anti-human PD-1 or PD-L1 antibodies, 1C3 also greatly potentiated anti-tumor responses compared to monotherapy of each agent. Analysis of tumor-infiltrating lymphocytes (TIL) demonstrated that 1C3 treatment significantly increased the ratio of effector T cells to Tregs in the TME. Moreover, 1C3 was well tolerated in mice, and no adverse effects were observed even at high doses (e.g. 100 mg/kg). In conclusion, these data demonstrate that 1C3 is a novel anti-human TNFR2 non-blocking agonist antibody with effective anti-tumor efficacy and favorable safety. 1C3 may exert these effects by regulating Treg and effector T cell activities.

Citation Format: Yongfei Yang, Shuzhen Cao, Wenjiao Zhang, Jing Zhang, Zhe Shao, Xueyuan Jiang, Qingya Duan, Maopeng Tian, Sen Mei, Zhaoxue Yu. 1C3, a novel non-blocking anti-human TNFR2 antibody, promotes effector T cell responses and demonstrates potent anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6436.
Abstract 6454: Metabolic obesity phenotype and risk of obesity-related cancers in the women's health initiative
Authors: Karra Prasoona , Winn Maci , Anderson Garnet , Haaland Benjamin , Shadyab Aladdin H. , Neuhouser Marian , Seguin-Fowler Rebecca , Thomson Cynthia A. , Coday Mace , Wactawski-Wende Jean , Stefanick Marcia L. , Zhang Xiaochen , Cheng Ting-Yuan David , Karanth Shama , Sun Yangbo , Saquib Nazmus , Pichado Margaret , Jung Su Yon , Tabung Fred , Summers Scott A. , Holland William L. , Jalili Thunder , Gunter Marc , Hardikar Sheetal , Playdon Mary C.
Published: June 2023

Introduction: Body mass index (BMI) may misclassify obesity-related cancer (ORC) risk since metabolic dysfunction can exist at any BMI. We measured the association of metabolic dysfunction, independent of BMI with risk of ORC.

Methods: Study included 60% white and 34% black Women’s Health Initiative participants with available baseline cardiovascular disease biomarkers. Metabolic obesity phenotypes were classified as normal weight with no metabolic dysfunction (NWNMD), normal weight with metabolic dysfunction (NWMD), overweight/obese with no metabolic dysfunction (OONMD) and overweight/obese with metabolic dysfunction (OOMD) when defined by Wildman and ATP III criteria. We performed a Cox proportional hazards regression with death as a competing risk adjusting for confounders.

Results: Defining by Wildman and ATP III criteria, 17.0%, 6.5%, 31.0%, 45.0%; and 22.2%, 2.4%, 41% and 35% were NWNMD, NWMD, OONMD and OOMD respectively. The risk of all ORC combined was elevated among NWMD (HR 1.12, 95% CI: 0.90-1.39), OONMD (HR 1.15, 95% CI: 1.00-1.32) and OOMD (HR 1.35, 95% CI: 1.18-1.54) when compared to NWNMD using Wildman criteria. Results were similar when defined by ATP III, hs-CRP and HOMA-IR. When stratified by cancer type, NWMD were at increased risk of colorectal cancer when compared to NWNMD (HR 1.70, 95% CI: 1.02-2.82). Individuals who were overweight/obese were at increased risk of cancer for all cancer types though the effect estimates were lower among OONMD phenotype.

Conclusions: NWMD and OOMD individuals are at increased risk of all ORC combined, independent of BMI status when compared to individuals with NWNMD.

Citation Format: Prasoona Karra, Maci Winn, Garnet Anderson, Benjamin Haaland, Aladdin H. Shadyab, Marian Neuhouser, Rebecca Seguin-Fowler, Cynthia A. Thomson, Mace Coday, Jean Wactawski-Wende, Marcia L. Stefanick, Xiaochen Zhang, Ting-Yuan David Cheng, Shama Karanth, Yangbo Sun, Nazmus Saquib, Margaret Pichado, Su Yon Jung, Fred Tabung, Scott A. Summers, William L. Holland, Thunder Jalili, Marc Gunter, Sheetal Hardikar, Mary C. Playdon. Metabolic obesity phenotype and risk of obesity-related cancers in the women's health initiative [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6454.
Abstract 6542: Detection of androgen receptor splice variants from clinical sequencing
Authors: Hwangbo Suhyun , Lee Sungyoung , Kim Sheehyun , Yun Hongseok
Published: June 2023

Androgen receptor (AR) splice variants (AR-Vs) have been widely studied on its important role in prostate cancer (PC) progression. In particular, AR-Vs lacking ligand binding domains, such as AR splice variant 7 (AR-v7) that links exon 3 and cryptic exon 3 or AR variant that losses exons 5 to 7 (ARv567es), have been demonstrated as one of the resistant mechanisms to androgen deprivation therapy in PC. Despite its clinical importance, the current algorithms to detect fusions such as TopHat-Fusion have limitation to detect intra-chromosomal rearrangements including AR-Vs. In this respect, we propose a novel approach to identify AR-Vs from targeted RNA sequencing (RNA-seq) dataset. In short, our two-step approach first gathers soft-clipped or divided reads that are adjacent to the splicing sites of AR-Vs and then yields the number of splitting reads that support the existence of AR-Vs. Next, the algorithm selects the paired-end reads whose one side and the other side are mapped to the preceding and following exons, respectively. The final number of spanning reads are calculated following to the removal of low-quality reads. We validated the proposed approach using two large-scale, independent RNA-seq datasets of PC samples: 34 samples from Seoul National University Hospital (SNUH) and 558 samples from The Cancer Genome Atlas (TCGA) dataset. Overall, our approach successfully identified samples with putative AR-Vs, including AR-v7 and ARv567es. In addition, our approach successfully detected AR-v7 with reliable number of supporting reads, from the RNA-seq dataset of AR-v7-expressing cell line. Finally, statistical analyses of 34 SNUH PC patients suggested potential detection threshold in clinical sequencing settings (at least 10 split reads and 1% proportion). In the SNUH dataset, The AR-v7 positive group, which included 6 patients (17.1%) with values above the threshold, had higher AR-v7 expression levels than the negative group (p=4.8E-04). The similar pattern was also found in the analysis of TCGA dataset. Owing to its flexibility, we also confirmed that the developed approach can be used to detection of other AR-Vs, including AR-v3 and ARv567es. We expect that the further in-depth analyses including larger samples and clinical outcomes can discover clinical applicability of AR-Vs.

Citation Format: Suhyun Hwangbo, Sungyoung Lee, Sheehyun Kim, Hongseok Yun. Detection of androgen receptor splice variants from clinical sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6542.
Abstract 6636: Metformin suppresses tumor immune evasion through downregulation of PD-L1 expression in cancer cells and PD-1 expression in NK cells and T cells
Authors: Park Su Hwan , Lee Jong-Ho
Published: June 2023

Up-regulation of programmed cell death 1 (PD-1) receptor on T cells and its ligand (PD-L1) on cancer cells is reported in various tumor microenvironment, and the interaction between PD-L1 and PD-1 inhibits the cytotoxic activity of NK cells and T cells, allowing cancer cells to evade immune destruction. Extensive studies suggested that metformin, the most widely used first-line drug for treatment of type 2 diabetes, had a potential efficacy of enhancing anti-tumor immune response. However, the detail mechanisms underlying the efficacy are unknown. Here, we showed that metformin decreases AKT-mediated β-catenin S552 phosphorylation and β-catenin transactivation in an AMPK activation-dependent manner, resulting in reduced CD274 transcription in cancer cells. In the cocultures of cancer cells with NK cells or T cells, cancer cells induced PD-1 protein stability and expression in the NK cells and T cells, these PD-1 upregulation was also observed in infiltrated NK cells and T cells in vivo mouse tumors. Metformin reduced the half-life and expression of PD-1 in an AMPK activation-dependent manner in vitro and in vivo. These inhibitory roles of metformin in PD-L1 expression in cancer cells and in cancer cell-induced PD-1 expression in NK cells and T cells led to ameliorated the cancer cell-reduced cytotoxic activity of NK cells and T cells and inhibited tumor growth in vivo models. These findings suggest that metformin may act as both PD-L1 and PD-1 blockades within tumor microenvironment, providing a mechanistic insight for the efficacy of metformin to improve immunotherapy in human cancer.

Citation Format: Su Hwan Park, Jong-Ho Lee. Metformin suppresses tumor immune evasion through downregulation of PD-L1 expression in cancer cells and PD-1 expression in NK cells and T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6636.
Abstract 6645: A systematic approach for detection of tumor genomic alterations which alter the immune microenvironment
Authors: Vella Raven , Hoskins Emily L. , Reeser Julie W. , Wing Michele R. , Samorodnitsky Eric , Paruchuri Anoosha , Smith Amy , Stein Leah , Dao Thuy , Risch Zachary , Roychowdhury Sameek
Published: June 2023

Objectives: Leveraging whole exome and transcriptome data from large cancer datasets, we sought to develop a computational approach to nominate candidate genomic alterations that are associated with hot or cold tumor. We anticipate this tool will identify candidate novel drug targets and biomarkers for immunotherapy in cancer.

Methods: The user selects a gene of interest (GOI) as input for the pipeline. The pipeline searches TCGA (The Cancer Genome Atlas) mutation and structural variant outputs to collect hits in the gene of interest. Each sample with an alteration in the GOI is annotated with the alteration’s predicted biological function (loss-of-function or LOF vs gain-of-function or GOF) using gene set enrichment analysis and pathway analysis from RNA sequencing. After annotating by sample, the alterations themselves are scored and recorded. Next, Quantiseq and Trust4 are employed to estimate features of the immune microenvironment in both the LOF and GOF samples. These results along with cancer type distributions, genomic position maps, and oncoplots are delivered to the user for further investigation. We are creating an online application to provide these results to the public. Simultaneously, a command line tool is being developed for application to any data source.

Results: We will present findings for the pipeline using positive controls including genomic alterations in ADAR, MTAP, IDH1, and CD274 (PD-L1). Preliminarily, we identified candidate GOF mutations (n=81) in PD-L1 that have inflamed tumor immune microenvironments like those of samples with known GOF structural variations in PD-L1. These samples have an overall “hot” immune microenvironment with significantly (p<.05) increased M1 macrophages, increased expression of IFN-gamma, and decreased M2 macrophages.

Conclusion: Currently there are no systematic approaches to interrogate individual genomic alterations and their association with an altered immune microenvironment. Our approach is pragmatic for any gene of interest, can be applied to any database, and identifies clinically actionable targets. This approach will identify heretofore unstudied connections between genomic alterations and immune microenvironment and support personalized drug development strategies in immuno-oncology.

Citation Format: Raven Vella, Emily L. Hoskins, Julie W. Reeser, Michele R. Wing, Eric Samorodnitsky, Anoosha Paruchuri, Amy Smith, Leah Stein, Thuy Dao, Zachary Risch, Sameek Roychowdhury. A systematic approach for detection of tumor genomic alterations which alter the immune microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6645.
Supplementary Figure 9 from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

Pearson's correlation coefficients analysis between immuno-PET signals and survival in ICI and control groups.
Supplementary Figure 8 from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

MRI analysis of tumor volume prior to immuno-PET.
Supplementary Figure 3 from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

CD69 analysis in single-cell RNA-seq data.
Supplementary Figure 2 from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

CD69 validation by tissue staining.
Supplementary Figure 10 from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

MRI analysis of tumor volume before and after treatment.
Supplementary Figure 1 from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

CD69 is upregulated upon T-cell activation.
Data from Immuno-PET Imaging of CD69 Visualizes T-Cell Activation and Predicts Survival following Immunotherapy in Murine GBM.
Authors: Nisnboym Michal , Vincze Sarah R. , Xiong Zujian , Sneiderman Chaim T. , Raphael Rebecca A. , Li Bo , Sever ReidAnn E. , Jaswal Ambika P. , Day Kathryn E. , Latoche Joseph D , Foley Lesley M , Karimi Hanieh , Hitchens T. Kevin , Agnihotri Sameer , Hu Baoli , Rajasundaram Dhivyaa , Anderson Carolyn J , Blumenthal Deborah T. , Pearce Thomas M. , Uttam Shikhar , Nedrow Jessie R. , Panigrahy Ashok , Pollack Ian F , Lieberman Frank S , Drappatz Jan , Raphael Itay , Edwards Wilson B. , Kohanbash Gary
Published: June 2023

Abstract
Glioblastoma (GBM) is the most common and malignant primary brain tumor in adults. Immunotherapy may be promising for the treatment of some GBM patients, however, there is a need for non-invasive neuroimaging techniques to predict immunotherapeutic responses. The effectiveness of most immunotherapeutic strategies requires T-cells activation. Therefore, we aimed to evaluate an early marker of T-cell activation, CD69, for its use as an imaging biomarker of response to immunotherapy for GBM. Herein, we performed CD69 immunostaining on human and mouse T-cells following in vitro activation and post immune-checkpoint inhibitors (ICI) in an orthotopic syngeneic mouse glioma model. CD69 expression on tumor-infiltrating leukocytes was assessed using single-cell RNA sequence (scRNA-seq) data from recurrent GBM patients receiving ICI. Radiolabeled CD69 antibody (Ab) positron emission tomography/computed tomography (PET/CT) imaging (CD69 immuno-PET) was performed on GBM-bearing mice longitudinally to quantify CD69 and its association with survival following immunotherapy. We show CD69 expression is upregulated upon T-cell activation and on tumor-infiltrating lymphocytes (TILs) in response to immunotherapy. Similarly, scRNA-seq data demonstrated elevated CD69 on TILs from ICI-treated recurrent GBM patients as compared with TILs from control cohorts. CD69 immuno-PET studies showed a significantly higher tracer uptake in the tumors of ICI-treated mice compared with controls. Importantly, we observed a positive correlation between survival and CD69 immuno-PET signals in immunotherapy-treated animals and established a trajectory of T-cell activation by virtue of CD69-immuno-PET measurements. Our study supports the potential use of CD69 immuno-PET as an immunotherapy response assessment imaging tool for GBM patients.
PENGARUH KECANDUAN PENGGUNAAN SMARTPHONE TERHADAP HASIL SCHIRMER TEST PADA MAHASISWA FK UPH TAHUN 2019
Authors: Irma Josiah , Angelina Jennifer
Published: June 2023

Pendahuluan : Penggunaan gadget terutama smartphone telah menjadi kebutuhan dasar bagi manusia. Berdasarkan penelitian yang telah dilakukan pada tahun 2019, pengguna gadget mayoritas adalah perempuan dengan persentase 71% dan sebanyak 2/5 dari mahasiswa menggunakan gadget selama 1-3 jam setiap harinya. Prevalensi populasi yang mengalami mata kering di Asia Tenggara memiliki variasi antara 20% - 52,4%. Pengguna smartphone dapat mengalami computer vision syndrome yang dapat menyebabkan mata kering. Salah satu pemeriksaan fisik yang dapat dilakukan adalah menggunakan schirmer test. Penelitian ini bertujuan untuk mengetahui apakah terdapat pengaruh kecanduan penggunaan smartphone terhadap hasil schirmer test pada mahasiswa Fakultas Kedokteran Universitas Pelita Harapan. Metode Penelitian : Penelitian ini menggunakan metode studi observasional dengan desain potong lintang pada 119 responden mahasiswa FK UPH yang menggunakan smartphone sehari-hari. Hasil Analisis Data : Data didapatkan dari 130 responden menggunakan kuesioner SAS-SV serta schirmer test. Data diolah menggunakan uji chi square. Penelitian ini menunjukkan adanya hasil yang signifikan antara kecanduan smartphone terhadap hasil schirmer test pada mahasiswa FK UPH dengan p value 0.001. Nilai odds ratio yang didapatkan sebesar 3.989 dan confidence interval 95% sebesar 1.744-9.124. Simpulan : Terdapat hubungan positif yang signifikan antara kecanduan penggunaan smartphone terhadap hasil schirmer test pada mahasiswa FK UPH.
KARAKTERISTIK DAN PENANGANAN TRAKHOMA PASIEN RAWAT JALAN DI RS. MATA PASURUAN PERIODE OKTOBER 2019 – JANUARI 2021
Authors: Rizqillah Nur
Published: June 2023

Pendahuluan: Trakhoma merupakan penyebab kebutaan ketiga setelah katarak dan glaukoma di seluruh dunia. Trakhoma penyebab gangguan penglihatan bagi sekitar 1,9 juta orang di seluruh dunia sekitar 1,4% diantaranya mengalami buta total. Di Indonesia prevalensi trakhoma yang menjadi penyebab kebutaan sekitar 0,98%. Penyakit trakhoma disebabkan oleh infeksi Chlamydia Trachomatis. Tujuan: Saat ini belum ada penelitian mengenai karakteristik dan pengobatan trakhoma di Pasuruan. Tujuan penelitian ini adalah untuk mengetahui karakteristik dan penanganan trakhoma pada pasien rawat jalan di Rumah Sakit Mata Pasuruan periode Oktober 2019 hingga Januari 2021. Metode: Penelitian yang digunakan adalah penelitian deskriptif diambil data retrospektif dengan metode cross sectional pada bulan Oktober 2019 hingga Januari 2021. Teknik pengambilan sampel dilakukan dengan cara total sampling. Data yang memenuhi kriteria inklusi selanjutnya dianalisis. Hasil: Dari 38 kasus trakhoma di Rumah Sakit Mata Pasuruan pada periode Oktober 2019-Januari 2021 didapatkan kasus terbanyak pada jenis kelamin perempuan (68,4%) dengan kategori usia 11-20 tahun (65,8%), trakhoma ODS trakhoma IIA jenis yang sering muncul (63,2%) dengan sebagian besar gejala muncul pada oculli dekstra sinistra (86,9%) dan penanganan dengan pemberian ointment oxytetracycline (55,3%). Simpulan : Secara umum trakhoma terbanyak pada jenis kelamin perempuan dengan usia 11-20 tahun, derajat yang sering muncul yaitu ODS trakhoma IIA dengan posisi gejala pada oculli dekstra sinistra, serta penanganan dengan pemberian ointment oxytetracycline. Penelitian ini merupakan hasil objektivitas peneliti terhadap kasus trakhoma, dan tidak ada maksud lain dari peneliti selain mendapatkan luaran hasil penelitian mengenai trakhoma.
IMMEDIATE MANAGEMENT OF OCULAR BEE STING RESULTS IN BETTER VISUAL OUTCOME – A CASE REPORT
Authors: Siska Siska , Pamudji Nadia Elena
Published: June 2023

Introduction: Ocular bee stings is an uncommon environmental eye injury that can result in various ocular complications. Due to the rarity of cases and conatroversial management, there are only a few literatures reported. Unfortunately, delay in management will be fatal for patients. Purpose: To highlight early management and complication of ocular bee sting case Case Presentation: A 24-year-old man presented to our hospital with chief complaint pain and blurred vision on his right eye in the last 2 days ago after being stung by bee. A part of the sting was already revoked, and the eye was rinsed off with tap water. The visual acuity on the right eye was hand movement with corneal edema, and bee sting track seen at the cornea. The anterior chamber was covered with hypopion and fibrins. An emergency bee sting extraction was quickly performed. We found the sting pierced in a diagonal way to the iris. We injected triamcinolone acetate intracamerally to make sure the exact location and how deep the sting embedded. The visual acuity improved to 6/30 a day after the surgery, but the cornea was melted and anterior uveitis still discovered, with increasing intraocular pressure. Topical and systemic corticosteroid, oral acetazolamide, topical antibiotic, and atropine were prescribed. The last condition, 2 weeks after extraction, best corrected visual acuity became 6/12 and no anterior chamber reaction discovered with corneal scarring.
PROGRESIVITAS DAN TATALAKSANA KERATOKONUS SETELAH SATU TAHUN TINDAKAN CORNEAL CROSS-LINKING
Authors: Haryono Aditia Apriyanto , Satari Karmelita , Sirait Susanti Natalya , Musa Ine Renata
Published: June 2023

Pendahuluan : Keratokonus merupakan kelainan ektasia kornea bersifat progresif tanpa inflamasi dengan penipisan dan penonjolan kornea yang menyebabkan astigmatisme ireguler. Diagnosis keratokonus ditegakan berdasarkan anamnesis, pemeriksaan oftamologi, dan pemeriksaan topografi. Tatalaksana keratokonus berupa kacamata, lensa kontak dan corneal cross-linking. Progresivitas keratokonus dapat diturunkan melalui tindakan corneal cross-linking. Tujuan : Untuk melaporkan progresivitas dan tatalaksana keratokonus setelah satu tahun tindakan corneal cross-linking. Laporan Kasus : Perempuan usia 30 tahun datang ke Rumah Sakit Mata Cicendo dengan keluhan buram kedua mata sejak 4 bulan yang lalu. Riwayat corneal cross-linking satu tahun yang lalu. Riwayat penggunaan kacamata 5 bulan yang lalu. Riwayat menggosok mata dan alergi. Pemeriksaan tajam penglihatan mata kanan 3/60 dengan pin hole 0,2 dan mata kiri 2/60 dengan pin hole 0,2. Tajam penglihatan terbaik dengan koreksi mata kanan S-5,00 C-5,00 x 180 sebesar 1,0 dan mata kiri S-6,00 C-6,00 x 170 sebesar 1,0. Pemeriksaan tomografi pentacam dibandingkan sebelum dan setelah tindakan cross-linking. Terdapat penurunan ketebalan kornea setelah satu tahun corneal cross-linking mata kanan 4% dan mata kiri 3,2%. Terdapat peningkatan nilai Kmax pada mata kanan 0,4D sedangkan mata kiri 2,6D. Simpulan : Tomografi pentacam dapat menilai progresivitas keratokonus. Tindakan corneal cross-linking dapat menurunkan progresivitas keratokonus. Tatalaksana keratokonus berdasarkan pada derajat keratokonus.
KARAKTERISTIK DAN PENANGANAN PENDERITA SINDROM RUBELA KONGENITAL DI PUSAT MATA NASIONAL RUMAH SAKIT MATA CICENDO
Authors: Kuntorini Mayasari Wahyu , Memed Feti Karfiati , Irfani Irawati , Oktarima Primawita , Caesarya Sesy
Published: June 2023

Introduction : Congenital rubella syndrome is a collection of symptoms caused by rubella virus infection during pregnancy. Congenital rubella syndrome often associated with ocular and systemic disease that are important for early diagnosis and management to prevent visual deprivation. Purpose : To determine the characteristic, clinical manifestation and management associated with congenital rubella syndrome in Cicendo Eye Hospital National Eye Center during January 2017 – December 2020. Methods :This was retrospective observational study and conducted from January 2017 – December 2020. The medical records of patients diagnosed with laboratory confirmed congenital rubella syndrome were included. We studied the demographic profile, associated ocular and systemic manifestation, and the management. Results : The study included 137 eyes of 84 patients with congenital rubella syndrome. Median duration of postoperative follow up was 9 months. Most patients were below 6 months of age and most patients were presented with bilateral manifestation. Ocular manifestations were congenital cataract, congenital glaucoma and pigmentary retinopathy. Non ocular manifestations were congenital heart disease, hearing impairment, microcephaly and developmental delay. Irrigation aspiration with primary posterior capsulotomy and anterior vitrectomy without intraocular lens implantation was the most frequent technique. Visual axisopacity was complication that were found after surgery. Conclusions :There were various ocular and systemic manifestation of congenital rubella syndrome. Surgical management were performed in all cases with irrigation aspiration with primary posterior capsulotomy and anterior vitrectomy was the most frequent technique. Keywords : Congenital rubella syndrome, ocular and non ocular manifestation, cataract and glaucoma surgery