Lymphocyte Tumor Research Articles

Normalization of Tumor Vessels by Lenvatinib-Based Metallo-Nanodrugs Alleviates Hypoxia and Enhances Calreticulin-Mediated Immune Responses in Orthotopic HCC and Organoids.

Immunocheckpoint inhibitors combined with Lenvatinib is the first line treatment for hepatocellular carcinoma (HCC), but their potency is hampered by the low response rate and adverse events. Herein, a targeted therapeutic strategy through the coassembly of Lenvatinib, Adriamycin, Fe3+ ion, and a natural polyphenol (metallo-nanodrugs) is presented by coordination effect for potentiating tumor vascular normalization and systematic chemo-immunotherapy to effectively inhibit the progression of HCC in both orthotopic model and patients-derived organoids. In mice with orthotopic HCC, the obtained metallo-nanodrugs efficiently increase the drug accumulation in orthotopic tumors and can respond to acidic tumor environment. The promotion of tumor vascular normalization by metallo-nanodrugs is observed, which enhances the infiltrating T lymphocytes in tumor, and reinforces the calreticulin-mediated antitumor immunity through alleviating hypoxia, reducing regulatory T cells, and down-regulating PDL1 expression of tumors. The excellent therapeutic efficiency with complete remission of orthotopic tumors (3/6) and long-term survival of mice (4/6, 42 days) are also achieved. Furthermore, the excellent therapeutic effect of metallo-nanodrugs is also validated in 5 patient-derived organoids, and hence can provide a marvelous systemic chemo-immunotherapy strategy for enhancing HCC treatment.

Abstract 6389: Differential role of CD8 cells and CD4 cells in anti-tumor immunity triggered by tumor cell apoptosis using HSVtk/GCV system

Abstract Immune cells can recognize tumor antigens and elicit an adaptive immune response, potentially resulting in tumor regression. Drug-induced dead tumor cells have been reported to enhance anti-tumor immunity, but results are conflicting, possibly due to the immunosuppressive effects of the cytotoxic drugs. To investigate whether apoptotic tumor cells trigger anti-tumor immunity independent of anti-cancer treatment, local immune responses were evaluated after direct induction of tumor cell apoptosis using a herpes simplex virus thymidine kinase (HSV-tk)/ganciclovir (GCV) system. The inflammatory cytokines IFNγ and IL-12β and anti-inflammatory factor PD-1 and TGFβ1 were upregulated at the tumor site after apoptosis induction. HSV-tk/GCV-induced tumor cell apoptosis resulted in suppression of tumor growth in vivo and promoted the infiltration of CD8 and CD4 T lymphocytes in tumor. To explore the role of T cells after induction of tumor cell death, GCV was administered together with anti-CD8 or anti-CD4 depleting antibodies to syngeneic HSV-tk transfected tumor-bearing mice. CD8 T cell depletion abrogated the anti-tumor efficacy of GCV, indicating that tumor regression was mainly dependent on CD8 T cells. This anti-tumor immunity was enhanced by anti-PD-1 antibody injection. On the contrary, CD4 T cell depletion further inhibited tumor growth than GCV alone, suggesting the potential role of CD4 T cells in suppressive tumor immunity. To elucidate this immunological mechanism, tumor tissues were evaluated after tumor cell apoptosis and CD4 T cell depletion. Foxp3, a regulatory T cell marker, was decreased after CD4 T cell depletion. In addition, Arg1, a marker of immunosuppressive myeloid cells, was significantly downregulated, which was expressed on tumor-associated macrophages (TAMs) and monocytic myeloid-derived suppressor cells (mMDSCs). Collectively, these findings indicate that tumor cell apoptosis accelerates CD8 T cell-dependent anti-tumor immunity and CD4 T cell-mediated Arg1 production from TAMs and mMDSCs. Citation Format: Sho Umegaki, Hidekazu Shirota, Tomoyuki Iwasaki, Yuki Kasahara, Chikashi Ishioka. Differential role of CD8 cells and CD4 cells in anti-tumor immunity triggered by tumor cell apoptosis using HSVtk/GCV system [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 6389.

Abstract 5399: Deep learning-based ensemble model using H&E images for the prediction of KRAS G12C mutations in non-small cell lung cancer

Abstract Background: As the KRAS G12C mutation became targetable in non-small cell lung cancer (NSCLC), tissue based KRAS mutation test is now an essential practice for the treatment decision. Recently, predicting KRAS mutations using deep-learning models with H&E images to potentially increase the pre-test probability has been reported with modest performance. Herein, we conducted a novel approach to improve the performance of KRAS G12C prediction based on an ensemble model trained not solely on H&E images, but also with multi-layered semantic content produced by a pre-trained artificial-intelligence (AI) analyzer, Lunit SCOPE IO. Methods: The Cancer Genome Atlas LUAD and LUSC (TCGA-Lung) samples were used for model development. A self-supervised vision transformer was used to extract deep features from raw H&E images; and an AI-based pathology profiling analyzer extracted semantic contents such as the spatial information of tumor cells, lymphocytes, cancer epithelium, and cancer stroma. A set of classifiers was trained based on the two features, and the ensemble of these features was used to improve robustness. The final model was evaluated through cross-validation and assessed on independent NSCLC samples from Samsung Medical Center (SMC) who tested KRAS mutation by various methods including whole exome sequencing or target sequencing. Results: TCGA-Lung dataset (n = 930) includes 150 (16.1%) KRAS driver mutations, and 62 (6.7%) KRAS G12C. The best cross-validation performances of the models predicting KRAS G12C, measured by mean area-under-the-curve (AUC), were 0.768 trained by only H&E images (HE-only), 0.714 by AI semantic content (AISC) with MLP classifier (AI-MLP), and 0.697 by AISC with random forest (AI-RF), respectively. An ensemble of the three models showed an increased AUC of 0.787 in TCGA-Lung by cross-validation. These models were applied to an independent SMC dataset (n = 363), including 54 (14.9%) KRAS driver mutations, and 22 (6.1%) KRAS G12C. The mean AUC to predict KRAS G12C by HE-only, AI-MLP, and AI-RF models were 0.599, 0.644, and 0.678, respectively, implying limited robustness. However, the AUC of an ensemble of the 3 models was 0.745 in the SMC dataset, showing 71.0% sensitivity and 72.7% specificity. Similar results were observed regardless of the KRAS testing method (TruSight Oncology 500 panel; n = 249; AUC 0.787, other tests; n = 114; AUC 0.720), the tissue size (surgical resection; n = 138; AUC 0.724, biopsy n = 225; AUC 0.763), and histology (excluding squamous cell carcinoma, n = 286; AUC 0.697). Conclusions: An AI-based ensemble model combining H&E images with semantic contents extracted from pre-developed AI model significantly improved the accuracy and the robustness of KRAS G12C mutation prediction using H&E sample in NSCLC. Citation Format: Sehhoon Park, Jongchan Park, Minuk Ma, Hyun-Ae Jung, Jong-Mu Sun, Yoon-La Choi, Jin Seok Ahn, Myung-Ju Ahn, Sanghoon Song, Gahee Park, Sukjun Kim, Huijeong Kim, Seunghwan Shin, Chan-Young Ock, Se-Hoon Lee. Deep learning-based ensemble model using H&E images for the prediction of KRAS G12C mutations in non-small cell lung cancer. [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 5399.

Abstract 3231: Temporal changes in intratumoral and systemic lymphocytes in response to short and long course radiotherapy regimens in locally advanced rectal cancer

Abstract Background. Two pre-operative radiotherapy (RT) regimens are in common use for patients (pts) with locally advanced rectal cancer (LARC): Conventionally fractionated, long course chemoradiation (LCCRT, 25x1.8Gy with concomitant chemotherapy) or hypofractionated short-course RT (5X5Gy) followed by systemic chemotherapy (SCRT-C). RT is thought to induce immune responses to cellular damage, but the understanding of their evolution and relation to fractionation is limited. We are conducting a serial biospecimen collection in such pts with baseline, 2, 6 and 12 week (wk) sampling. We report serial measurement of tumor and peripheral blood lymphocyte responses for each regimen. Method. Multiplex immunofluorescence quantified tumor infiltrating lymphocytes (CD8, FOXP3). Routine diagnostic flow cytometry quantified circulating lymphocytes. A multiplex ELISA quantified cytokines in blood plasma. Bulk RNA-sequencing (QuantSeq) quantified gene expression within the tumor. Result. We report results for 20pts who received RT for stage III/IV LARC (13 LCCRT, 7 SCRT-C). Biopsy results are available for 10pts, circulating lymphocytes for 20pts and peripheral blood cytokines for 16pts. In LCCRT patients (n=8), relative to baseline, tumor infiltrating cytotoxic (CD8+) T cells were uniformly decreased during (2wks, P<0.05) and directly after treatment (6wks, P<0.05) before returning to baseline at 12wks. T regulatory cells (FOXP3+) similarly significantly decreased at 2wks and 6wks but remained below baseline at 12wks (P<0.05). Circulating lymphocytes also fell at wk2 and wk6 after commencing LCCRT and had begun recovering by wk12(n=13; P<0.05). The concentrations of circulating interleukins secreted by (IL2, IL8) or which activate T-lymphocytes (IL2, IL15) were reduced in the circulation at wk2 (n= 9; P<0.05) and wk6 (NS). In SCRT-C patients, we noted an increase in CD8 T cells at 2wks in 2/2 pts, also reflected in gene expression data, and an increase in FOXP3 T cells in 1/2 pts. Circulating lymphocytes were similarly decreased at wk2 in both the SCRT-C (n=7) and LCCRT (n=13) pts, but this reduction was less marked within the SCRT-C cohort at wk6 (P<0.01) and wk12 (P<0.05) relative to LCCRT patients. IL2, IL8 and IL15 did not change during or after SCRT-C (n=7; NS). Conclusion. LCCRT caused a drop in T cells during treatment, whilst SCRT-C appears to induce intra-tumoral T cell responses from wk2 and abrogates systemic reactions to a lesser extent. These results require evaluation in a larger cohort but have implications for understanding how RT induces microenvironmental changes and impacts pelvic bone marrow. We show in vivo that SCRT may be more immunostimulatory in LARC, with implications for trials combining RT with immunotherapy. Citation Format: Lily V. Hillson, Ross K. McMahon, Kathryn A. Pennel, Jean A. Quinn, Leia Jones, Raheleh Amirkhah, Aula Ammar, Phimmada Hatthakarnkul, Annabelle Ferguson, Simon W. Milling, Alec McDonald, Philip D. Dunne, Joanne Edwards, Sean M. O'Cathail, Campbell S. Roxburgh. Temporal changes in intratumoral and systemic lymphocytes in response to short and long course radiotherapy regimens in locally advanced rectal cancer [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 3231.

Can Neutrophil to Lymphocyte Ratio, Platelet to Lymphocyte Ratio and Tumor Necrosis Factor Alpha Predict Lupus Nephritis Flares?

Can Neutrophil to Lymphocyte Ratio, Platelet to Lymphocyte Ratio and Tumor Necrosis Factor Alpha Predict Lupus Nephritis Flares?

Dissecting tumor lymphocyte infiltration to predict benefit from immune-checkpoint inhibitors in metastatic colorectal cancer: lessons from the AtezoT RIBE study

BackgroundTumor immune cells influence the efficacy of immune-checkpoint inhibitors (ICIs) and many efforts aim at identifying features of tumor immune microenvironment able to predict benefit from ICIs in proficient mismatch...

Staphylococcal enterotoxin B as DNA vaccine against breast cancer in a murine model.

Recently, many efforts have been made to treat cancer using recombinant bacterial toxins and this strategy has been used in clinical trials of various cancers. Therapeutic DNA cancer vaccines are now considered as a promising strategy to activate the immune system against cancer. Cancer vaccines could induce specific and long-lasting immune responses against tumors. This study aimed to evaluate the antitumor potency of the SEB DNA vaccine as a new antitumor candidate against breast tumors in vivo. To determine the effect of the SEB construct on inhibiting tumor cell growth in vivo, the synthetic SEB gene, subsequent codon optimization, and embedding the cleavage sites were sub-cloned to an expression vector. Then, SEB construct, SEB, and PBS were injected into the mice. After being vaccinated, 4T1 cancer cells were injected subcutaneously into the right flank of mice. Then, the cytokine levels of IL-4 and IFN-γ were estimated by the ELISA method to evaluate the antitumor activity. The spleen lymphocyte proliferation, tumor size, and survival time were assessed. The concentration of IFN-γ in the SEB-Vac group showed a significant increase compared to other groups. The production of IL-4 in the group that received the DNA vaccine did not change significantly compared to the control group. The lymphocyte proliferation increased significantly in the mice group that received SEB construct than PBS control group (p < 0.001). While there was a meaningful decrease in tumor size (p < 0.001), a significant increase in tumor tissue necrosis (p < 0.01) and also in survival time of the animal model receiving the recombinant construct was observed. The designed SEB gene construct can be a new model vaccine for breast cancer because it effectively induces necrosis and produces specific immune responses. This structure does not hurt normal cells and is a safer treatment than chemotherapy and radiation therapy. Its slow and long-term release gently stimulates the immune system and cellular memory. It could be applied as a new model for inducing apoptosis and antitumor immunity to treat cancer.

CX3CR1 deficiency-induced TIL tumor restriction as a novel addition for CAR-T design in solid malignancies

Advances in the understanding of the tumor microenvironment have led to development of immunotherapeutic strategies, such as chimeric antigen receptor Tcells (CAR-Ts). However, despite success in blood malignancies, CAR-T therapies in solid tumors have been hampered by their restricted infiltration. Here, we used our understanding of early cytotoxic lymphocyte infiltration of human lymphocytes in solid tumors invivo to investigate the receptors in normal, adjacent, and tumor tissues of primary non-small-cell lung cancer specimens. We found that CX3CL1-CX3CR1 reduction restricts cytotoxic cells from the solid-tumor bed, contributing to tumor escape. Based on this, we designed a CAR-T construct using the well-established natural killer group 2, member D (NKG2D) CAR-T expression together with overexpression of CX3CR1 to promote their infiltration. These CAR-Ts infiltrate tumors at higher rates than control-activated Tcells or IL-15-overexpressing NKG2D CAR-Ts. This construct also had similar functionality in a liver-cancer model, demonstrating potential efficacy in other solid malignancies.

Construction a new nomogram prognostic model for predicting overall survival after radical resection of esophageal squamous cancer.

Esophageal cancer is one of the deadliest malignancies in the world, and 5-year overall survival (OS) of esophageal cancer ranges from 12% to 20%. Surgical resection remains the principal treatment. The American Joint Commission on Cancer (AJCC) TNM (tumor, node, and metastasis) staging system is a key guideline for prognosis and treatment decisions, but it cannot fully predict outcomes. Therefore, targeting the molecular and biological features of each patient's tumor, and identifying key prognostic biomarkers as effective survival predictors and therapeutic targets are highly important to clinicians and patients. In this study, three different methods, including Univariate Cox regression, Lasso regression, and Randomforest regression were used to screen the independent factors affecting the prognosis of esophageal squamous cell carcinoma and construct a nomogram prognostic model. The accuracy of the model was verified by comparing with TNM staging system and the reliability of the model was verified by internal cross validation. Preoperative neutrophil lymphocyte ratio(preNLR), N-stage, p53 level and tumor diameter were selected to construct the new prognostic model. Patients with higher preNLR level, higher N-stage, lower p53 level and larger tumor diameter had worse OS. The results of C-index, Decision Curve Analysis (DCA), and integrated discrimination improvement (IDI) showed that the new prognostic model has a better prediction than the TNM staging system. The accuracy and reliability of the nomogram prognostic model were higher than that of TNM staging system. It can effectively predict individual OS and provide theoretical basis for clinical decision making.

Cancer cell membrane–encapsulated biomimetic nanoparticles for tumor immuno-photothermal therapy

Immune checkpoint blockade is one of the most promising approaches in cancer immunotherapy, but patient response rates are low due to low tumor immunogenicity. Herein, we proposed a biomimetic nanoparticle loaded with indocyanine green (ICG) and NLG919 (an effective IDO-1 enzyme inhibitor), which can enhance the immunogenicity of tumor through photothermal and increase the infiltration of cytotoxic T lymphocytes in tumor, thus enhancing the immunotherapy response. In a 4T1 bilateral tumor model, CFIN targeted to the tumor site, which mediated the superior photothermal effect and eliminated the primary tumor. More importantly, high temperature improved the immunogenicity of tumors and created a favorable niche for tumor-infiltrating lymphocytes (TILs) recruitment. At the same time, NLG919 was released to inhibit the activity of IDO-1 and prevented the degradation of tryptophan, thus reversing the microenvironment of immunosuppressed tumors and producing strong prevention of distant metastasis of tumors. Therefore, CFIN combines the advantages of photothermal therapy and immunotherapy to activate mature dendritic cells, initiate cytotoxic T lymphocytes, trigger the body’s innate and adaptive immune responses, eliminate primary tumors, and inhibit distal tumor growth. More importantly, CFIN can be used as a nanovaccine to provide long-term immune memory effect immunity and prevent tumor recurrence. This strategy provides a simple but promising approach for treating metastatic cancer and preventing tumors.

A bispecific peptide-polymer conjugate bridging target-effector cells to enhance immunotherapy.

Peptide-based immune checkpoint inhibitors exhibited remarkable therapeutic benefits although their application was hindered by quick blood clearance and low affinity with receptors. The modification of the peptides into artificial antibodies was an ideal platform to solve these problems, and one of the optional pathways was the conjugation of peptides with a polymer. More importantly, the bridging effect, mediated by bispecific artificial antibodies, could promote the interaction of cancer cells and T cells, which will benefit cancer immunotherapy. Herein, we prepared a bispecific peptide-polymer conjugate (octa PEG-PD1-PDL1) by simultaneously conjugating PD1-binding and PDL1-binding peptides onto 8-arm-PEG. octa PEG-PD1-PDL1 bridged T cells and cancer cells and thus enhanced T cell-mediated cytotoxicity against cancer cells. Meanwhile, the tumor-targeting octa PEG-PD1-PDL1 increased the infiltration of cytotoxic T lymphocytes in tumors and reduced their exhaustion. It effectively activated the tumor immune microenvironment and exerted a potent antitumor effect against CT26 tumor models with a tumor inhibition rate of 88.9%. This work provided a novel strategy to enhance tumor immunotherapy through conjugating bispecific peptides onto a hyperbranched polymer to effectively engage target-effector cells. This article is protected by copyright. All rights reserved.

Synthetic enforcement of STING signaling in cancer cells appropriates the immune microenvironment for checkpoint inhibitor therapy

Immune checkpoint inhibitors (ICIs) enhance anticancer immunity by releasing repressive signals into tumor microenvironments (TMEs). To be effective, ICIs require preexisting immunologically "hot" niches for tumor antigen presentation and lymphocyte recruitment. How the mutational landscape of cancer cells shapes these immunological niches remains poorly defined. We found in human and murine colorectal cancer (CRC) models that the superior antitumor immune response of mismatch repair (MMR)-deficient CRC required tumor cell-intrinsic activation of cGAS-STING signaling triggered by genomic instability. Subsequently, we synthetically enforced STING signaling in CRC cells with intact MMR signaling using constitutively active STING variants. Even in MMR-proficient CRC, genetically encoded gain-of-function STING was sufficient to induce cancer cell-intrinsic interferon signaling, local activation of antigen-presenting cells, recruitment of effector lymphocytes, and sensitization of previously "cold" TMEs to ICI therapy in vivo. Thus, our results introduce a rational strategy for modulating cancer cell-intrinsic programs via engineered STING enforcement to sensitize resistant tumors to ICI responsiveness.

Immunohistochemical profiling of the tumour microenvironment in borderline and malignant ovarian tumours in young women.

Ovarian cancer is a major cause of cancer-related deaths in women. Our previous study highlighted the interaction between cancer cells and the host immune response in solid cancers. The present study aimed to analyse the proportion, density and distribution of T and B lymphocytes within the tumour and surrounding stroma, and their prognostic significance in young women with borderline and malignant ovarian surface epithelial tumours. Full clinicopathological and outcome data were collected for 57 women aged <50 years diagnosed between January 2010 and December 2015. Representative tumour sections were stained for CD3 (T cells) and CD20 (B cells) and tumour-infiltrating lymphocytes (TILs) were scored following the TILs Working Group Recommendations and described as stromal, intra-tumoural, lymphoid aggregates and touching lymphocytes. Data were statistically analysed and the association with clinicopathological variables was assessed. The median age was 41 years and the most common histological type was serous carcinoma (n=21). The risk of malignancy index was a significant predictor of ovarian cancer diagnosis (P<0.05). A total of 15 out of 34 patients with cancer died. There was significantly greater stromal infiltration of CD3 and CD20 TILs (P=0.01 and P=0.03, respectively) and higher intratumoral CD20 expression in ovarian epithelial cancers compared with borderline tumours. The highest CD3 stroma count and density were observed in serous carcinoma, which also exhibited the highest numbers of CD3 and CD20 aggregates. There was no statistically significant difference between touching lymphocytes and tumour histological subtype. There was no significant association between TIL expression and patient survival. The count, distribution and density of T and B lymphocytes in ovarian tumours varied depending on tumour type and invasiveness. Their topographic distribution within the tumour and surrounding stroma did not impact prognosis in young women with ovarian cancer. TIL analysis in an older age group of women with ovarian tumours is ongoing to determine its potential prognostic significance.

The significance of spread through air spaces in the prognostic assessment model of stage I lung adenocarcinoma and the exploration of its invasion mechanism.

Spread through air spaces (STAS) is a crucial invasive mode of lung cancer and has been shown to be associated with early recurrence and metastasis. We aimed to develop a prognostic risk assessment model for stage I lung adenocarcinoma based on STAS and other pathological features and to explore the potential relationship between CXCL-8, Smad2, Snail, and STAS. 312 patients who underwent surgery at Harbin Medical University Cancer Hospital with pathologically diagnosed stage I lung adenocarcinoma were reviewed in the study. STAS and other pathological features were identified by H&E staining, and a prognostic risk assessment model was established. The expression levels of CXCL8, Smad2, and Snail were determined by immunohistochemistry. The nomogram was established based on age, smoking history, STAS, tumor lymphocyte infiltration, tissue subtype, nuclear grade, and tumor size. The C-index for DFS was (training set 0.84 vs validation set 0.77) and for OS was (training set 0.83 vs validation set 0.78). Decision curve analysis showed that the model constructed has a better net benefit than traditional reporting. The prognostic risk score validated the risk stratification value for stage I lung adenocarcinoma. STAS was an important prognostic factor associated with stronger invasiveness and higher expression of CXCL8, Smad2, and Snail. CXCL8 was associated with poorer DFS and OS. We developed and validated a survival risk assessment model and the prognostic risk score formula for stage I lung adenocarcinoma. Additionally, we found that CXCL8 could be used as a potential biomarker for STAS and poor prognosis, and its mechanism may be related to EMT.

Investigation of the effect of perineural invasion on survival rates in rectal tumors.

Purpose: Colorectal cancers compose the third most common cancer group leading to death, with increasing rates in developed countries after lung and breast cancers. Rectal cancers make up about 30% of all colorectal cancers. Although many factors affecting recurrence and survival in rectal cancer have been identified, the stage of the tumor is the most important prognostic factor. However, many additional parameters that can be associated with survival and affect the adjuvant treatment plan have been identified besides the stage. Perineural invasion (PNI) is one of the parameters that can be associated with survival and affect the adjuvant treatment plan in colorectal cancers. The aim of this study is to investigate the effects of various histopathological prognostic factors, including PNI, on disease-free survival, overall survival, and recurrence rates in patients with rectal cancer who underwent surgery. Materials and methods: The clinical records of patients diagnosed with rectal cancer and operated on at the Department of General Surgery, Faculty of Medicine, Pamukkale University between 2008-2014 were reviewed. To investigate the factors affecting disease-free survival, recurrence rates, and overall survival, the presence of PNI, serum CEA levels, cTNM stage, pT stage, pN stage, number of metastatic LNs, tumor budding status, histopathological grades, lymphocyte infiltration, and presence of desmoplasia were evaluated in patients. &#x0D; Results: A total of 124 patients were included in the study. 42 patients (33.9%) had PNI while 82 patients (66.1%) did not have PNI. Of the 45 patients with recurrence, 26 had PNI while 19 did not have PNI. Of the 79 patients without recurrence, 16 had PNI while 63 did not have PNI. Similarly, PNI was seen more frequently in deaths related to the disease. The 3-year average survival of patients with and without PNI was 55% and 89%, respectively (p

Abstract P6-01-47: Stromal tumor infiltrating lymphocytes and pathological complete response in patients with inflammatory breast cancer treated with neoadjuvant chemotherapy

Abstract Background: Inflammatory breast cancer (IBC) is a rare (1-5%), but aggressive form of breast cancer (BC), accounting for ~10% of BC mortality. In early setting (M0), standard of care is neoadjuvant chemotherapy (NACT), followed by surgery. Nevertheless, outcome is still relatively poor. Pathological complete response (pCR) after NACT is prognostic in BC in general, and can be predicted by a high percentage of stromal tumor infiltrating lymphocytes (sTIL) in the primary tumor. The predictive value of sTIL in IBC has only been sporadically investigated, often in smaller series. Our aim was to determine which variables, including sTIL, are associated with pCR and to determine the prognostic value of pCR in IBC in a large multicentric, retrospective cohort. Patients &amp; Methods: We included patients with IBC treated with NACT+/- anti-Human Epidermal growth factor Receptor 2 (HER2) therapy, followed by surgery from 10/1996 to 10/2021 in 7 different European hospitals. Clinicopathological variables were collected and central pathological review was performed, including sTIL scoring. This study focused on M0 cases. Considered clinicopathological variables were: age, histology, tumor grade, estrogen receptor status (ER), HER2 status, focality (unifocal vs not), and baseline locoregional nodal status (Table 1). Associations between pCR, clinicopathological variables and sTIL were assessed using Firth’s logistic regression models: Model 1 was adjusted for center, Model 2 additionally included all variables of interest. Similarly, linear regression was used to investigate the association between sTIL and clinicopathological features. Univariable and multivariable Cox regression was used to evaluate the role of pCR on disease free survival (DFS), distant recurrence free survival (DRFS) and overall survival (OS). DFS and DRFS were analyzed considering death without the respective event as competing risk. Results: 494 patients were included. The distribution according to receptor status was: ER-/HER2- (24.3%), ER+/HER2- (34.4%), ER+/HER2+ (13%) and ER-/HER2+ (20.2%). pCR rate was 26% and per receptor status: ER-/HER2- (28%), ER+/HER2- (10%), ER+/HER2+ (42%) and ER-/HER2+ (45%). pCR was associated with grade (G3 vs G1/2, OR =2.79 (1.70 − 4.74), p &amp;lt; .001), ER-status (positive vs negative, OR = 0.39 (0.26 − 0.60), p &amp;lt; .001) and HER2 status (positive vs negative, OR = 3.74 (2.43 − 5.81), p &amp;lt; .001) in Model 1. Only the association with HER2 status remained significant in Model 2 (OR = 5.34 (2.83 − 10.47), p &amp;lt; .001). sTIL was scored for 385 patients. Median sTIL was 5.3% [IQR 2.0%;16.7%] and according to receptor status: ER-/HER2- (10%), ER+/HER2- (2.5%), ER+/HER2+ (6.7%) and ER-/HER2+ (8.3%). Higher sTIL was associated with NST (p = .032), grade 3 (p = .015), and ER-negativity (p = .007) in Model 1. This was no longer significant in Model 2, but the direction of the trends was preserved. sTIL was associated with pCR (5% increment, OR = 1.13 (1.05 − 1.22), p = .002), but no longer after adjustment. No association between pCR and sTIL was found stratifying by receptor status. The median FU was 9.4 years and multivariable Cox regression models revealed that ER+ and HER2+ status and achieving pCR were significantly associated with better DFS, DRFS, and OS (Table 1). Conclusion: Our results indicate that patients with HER2+ tumors have a higher probability of achieving pCR and that pCR has an independent prognostic role in IBC. This is the largest IBC study with centrally scored sTIL, demonstrating that sTIL is associated with pCR but its role as an independent predictor of pCR is still not certain. Citation Format: Maxim De Schepper, Ha-Linh Nguyen, François Richard, Florence Lerebours, Roman Vion, Florian Clatot, Anca Berghian, Marion Maetens, Sophia Leduc, Edoardo Isnaldi, Chiara Molinelli, Matteo Lambertini, Frederica Grillo, Gabriele Zoppoli, Luc Dirix, Hilde Wuyts, Kevin Punie, Hans Wildiers, Chantal Remmerie, Ann Smeets, Ines Nevelsteen, Patrick Neven, Anne Salomon, Denis Larsimont, Caroline Duhem, Patrice Viens, François BERTUCCI, Elia Biganzoli, Peter Vermeulen, Giuseppe Floris, Christine Desmedt. Stromal tumor infiltrating lymphocytes and pathological complete response in patients with inflammatory breast cancer treated with neoadjuvant chemotherapy [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-01-47.

Abstract P2-23-17: Programmed death ligand-1 expression in triple-negative breast cancer from Nigeria

Abstract Introduction: The incidence of triple negative breast cancer (TNBC) in West Africa appears to mirror the higher incidence of the disease among African American women in the United States. However, there remains a paucity of molecular data on TNBC from sub-Saharan Africa, despite the emergence of effective immunotherapies. Methods: Consecutive patients diagnosed with invasive breast cancer between March 2018-Jan 2020 were identified from a prospective clinical database and paired biobank at Obafemi Awolowo University Teaching Hospital (OAUTH). All specimens were processed and fixed in formalin within 60 minutes of excision by a trained pathologist. Tissue sections (4μm thick) representative of tumor were selected for routine evaluation of estrogen, progesterone and human epidermal growth factor receptor-2 expression by immunohistochemistry. This was performed at OAUTH with adequate controls according to the ASCO/CAP guidelines and verified at an outside institution for quality assurance. Additional sections from the FFPE blocks of TNBC specimens were further stained using the Dako PharmDx 22C3 PD-L1 commercial assay according to manufacturer protocols at Dalhousie University. External on-slide controls included tonsil, PD-L1 negative TNBC, and PD-L1 positive TNBC. PD-L1 expression was scored using the combined positive score (CPS), which is the number of 22C3 staining tumour cells, lymphocytes, and macrophages divided by the number of viable tumour cells, multiplied by 100. The threshold for a positive result was a CPS of ≥10 as per manufacturer instructions and institutional protocol. Research ethics board approval as well as data and material transfer agreements between institutions was obtained for this study. Results: From 85 cases, 32 were TNBC (37.6%). The mean age and BMI were 49.6±SD 7.7 and 26.2±5.7, respectively. The majority of patients presented with locally advanced disease (64.2% Stage III, 14.2% Stage IV). Seventy-nine percent (78.5%) of patients received an average of five cycles of neoadjuvant chemotherapy (SD 2.3). From 32 TNBC specimens, 27 had available FFPE tissue blocks, and of those, 24 had interpretable PD-L1 IHC for inclusion in the analysis. A total of 37.5% (9/24) of cases demonstrated a CPS ≥10. Conclusions: Over a third of breast cancer specimens in this Nigerian cohort were triple negative, 37.5% of which had PD-L1 CPS scores of ≥10. These results suggest a large proportion of patients in Nigeria may benefit from access to immunotherapy. This is the first reported incidence of PD-L1 expression in breast cancer from sub-Saharan Africa. Citation Format: Olalekan Olasehinde, Funmilola Wuraola, Aleksandra Kajetanowicz, Gilllian Bethune, Marcia Edelweiss, Peter Ntiamoah, Oluwole Odujoko, Avinash Sharma, Victoria Mango, Peter Kingham, Olusegun Alatise, Gregory Knapp. Programmed death ligand-1 expression in triple-negative breast cancer from Nigeria [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-23-17.

VISTA H-Score Is Significantly Associated with a 5-Year DFS Rate in Oral Squamous Cell Carcinoma

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer in the world. Despite its prevalence, it is often recognized in advanced stages (III or IV) when it has already spread to local lymph nodes. In this study, we investigate the V-domain Ig suppressor of T cell activation (VISTA) as a potential prognostic factor in OSCC. Tissue samples were collected from 71 oral squamous cell carcinoma patients to determine protein expression levels (using immunochemistry and the semi-quantitative H-score method). Moreover, RT-qPCR was additionally performed in 35 patients. Clinical factors in our cohort study had no impact on VISTA expression. However, VISTA expression is largely correlated with Il-33 levels in tumor cells and lymphocytes and with PD-L1 in tumor cells. The impact of VISTA expression on overall survival (OS) is rather limited, but in the case of a 5-year survival rate, a significant association has been proven. VISTA seems to be a rather weak clinicopathological marker but needs further evaluation in the context of survival. In addition, the potential of VISTA combination with Il-33 or PD-L1 should be further investigated in OSCC.

Impact of Tumor-Infiltrating Lymphocytes on Disease Progression in Human Papillomavirus-Related Oropharyngeal Carcinoma.

We aim to explore the prognostic value of tumor-infiltrating lymphocytes (TILs) in the primary tumor and metastatic lymph nodes of patients with HPV(+)OPSCC. We hypothesize that TILS density at both sites is associated with disease-free survival in HPV(+)OPSCC. Matched case-control study among HPV(+)OPSCC patients who underwent intent-to-cure surgery. Cases developed locoregional or distant recurrence. Controls were matched based on age, sex, pathologic T, N, and overall stage, year of surgery, type of adjuvant treatment received, and the Adult Comorbidity Evaluation-27 (ACE-27) score. Single tertiary care center, May 2007 to December 2016. Tumoral TILs (tTILs) density was defined as % TILs; stromal TILs (sTILs) density was defined as absent/sparse or moderate/dense crowding. Associations between TILs and time to disease progression were assessed using Cox regression models. Forty-four case-control pairs (N = 88) were included: 42 (48%) AJCC pStage I, 39 (44%) pStage II, and 7 (8%) pStage III. tTILs density ≥10% (hazard ratio [HR] 0.41, 95% confidence interval [CI] 0.17-0.99, p = .048) and a moderate/dense sTILs density (HR 0.21, 95% CI 0.06-0.75, p = .016) in the primary tumor were significantly associated with decreased risk of progression. TILs density in the lymph node was associated with decreased risk of progressionbut did not reach statistical significance. The tTILs and sTILs density correlated strongly between the primary tumor and lymph node. Concordance between the pathologists' was moderate (60%-70%). In HPV(+)OPSCC, a higher density of tumoral and stromal TILs in the primary tumor and possibly the lymph node may predict a lower risk of disease progression.

Ultrasound targeted microbubble destruction-triggered nitric oxide release via nanoscale ultrasound contrast agent for sensitizing chemoimmunotherapy

Immunotherapy had demonstrated inspiring effects in tumor treatment, but only a minority of people could benefit owing to the hypoxic and immune-suppressed tumor microenvironment (TME). Therefore, there was an urgent need for a strategy that could relieve hypoxia and increase infiltration of tumor lymphocytes simultaneously. In this study, a novel acidity-responsive nanoscale ultrasound contrast agent (L-Arg@PTX nanodroplets) was constructed to co-deliver paclitaxel (PTX) and L-arginine (L-Arg) using the homogenization/emulsification method. The L-Arg@PTX nanodroplets with uniform size of about 300 nm and high drug loading efficiency displayed good ultrasound diagnostic imaging capability, improved tumor aggregation and achieved ultrasound-triggered drug release, which could prevent the premature leakage of drugs and thus improve biosafety. More critically, L-Arg@PTX nanodroplets in combination with ultrasound targeted microbubble destruction (UTMD) could increase cellular reactive oxygen species (ROS), which exerted an oxidizing effect that converted L-Arg into nitric oxide (NO), thus alleviating hypoxia, sensitizing chemotherapy and increasing the CD8 + cytotoxic T lymphocytes (CTLs) infiltration. Combined with the chemotherapeutic drug PTX-induced immunogenic cell death (ICD), this promising strategy could enhance immunotherapy synergistically and realize powerful tumor treatment effect. Taken together, L-Arg@PTX nanodroplets was a very hopeful vehicle that integrated drug delivery, diagnostic imaging, and chemoimmunotherapy.Graphical