Differences In Immune Composition Research Articles

Abstract A028: Disparities in suppressive immunity in Hispanic/Latina patients with hormone receptor positive breast cancer

Abstract In the US, breast cancer is the number one cancer diagnosed, and it is responsible for the most cancer related deaths among Hispanic/Latina (H/L) women. The mean age of diagnosis of breast cancer in H/L patients is 3.7 years younger than their Non-Hispanic White (NHW) counterparts. Despite being diagnosed with breast cancer at a younger age, the H/L population has a better overall survival rate across all breast cancer subtypes. This study investigates whether the immune response mounted against tumor cells in the H/L population differs from that mounted by NHW patients. We recruited and received consent from patients diagnosed with stage I- III (early stage), or stage IV/advanced (metastatic), hormone receptor positive (HR+) breast cancer. We collected blood from 66 patients (38 H/L, 28 NHW) across all stages of disease, and tumor tissue from 14 unmatched patients (7 H/L, 7 NHW), with early stage disease, at the time of tumor resection, none of whom had received neoadjuvant chemotherapy. We utilized multiparameter flow cytometry to analyze peripheral blood mononuclear cells (PBMCs), and 10X Visium for spatial transcriptomic and proteomic evaluation of patient’s tumor tissue to characterize differences in immune composition by race. We hypothesize that evaluation of intra-tumoral immune composition will reveal fewer immune suppressor cells and increased infiltration of cytotoxic immune cells in tumors from H/L patients. We also hypothesize evaluation of immune cells in circulation will mirror findings from within tumors. Suppressor immune cells, comprised of M2-polarized macrophages, myeloid derived suppressor cells (MDSCs), and T regulatory cells (Tregs) modulate T cell proliferation and cytotoxicity to affect anti-tumor immunity. We report a significant difference in the levels of both anti-tumoral and suppressive immune cells in the periphery within the H/L patient population when comparing early vs. metastatic HR+ breast cancer. This was not observed in the NHW group and there were no differences when comparing peripheral suppressor and T cell populations in H/L vs. NHW at any stage. Comparisons of intra-tumoral immune cell infiltration and differences in transcription profiles are forthcoming. The findings from this study will be some of the first to look at the immune composition in H/L women with HR+ disease, and could provide data to support larger studies leading to a paradigm shift in how we approach clinical trial design for immune checkpoint inhibition. Citation Format: Sabrina Carrel, Michelle Li, Batul Al-Zubeidy, Dominic Zavala, Edgar Gonzales, Aaron Baugh, Sofi Castanon, Robert Hsu, Dimitrios N Sidiropoulos, Yanling Ma, Michael Press, Evanthia T Roussos Torres. Disparities in suppressive immunity in Hispanic/Latina patients with hormone receptor positive breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr A028.

Efficacy of PD-1 checkpoint inhibitor therapy in melanoma and beyond: are peripheral T cell phenotypes the key?

Immunotherapy treatment strategies have proven effective in a limited portion of patients, where identifying responders from non-responders to treatment remains a challenge. While some indications can be drawn from invasive biopsies, we need more accessible methods for predicting response and better correlates of response prior to starting therapy. Recent work has identified differences in immune composition at baseline in peripheral blood from melanoma patients responding to PD-1 blockade treatment. Through flow cytometric analysis of T cell receptors, phenotypical features of CD8+ and CD4+ T cells and Tregs could allow for the stratification of treatment response. Analysing T cells within peripheral blood could potentially allow for the stratification of PD-1 treatment response prior to therapy in different cancer settings.

Abstract P416: Differences In Immune Cell Composition And Response Between Spontaneously Hypertensive Bph/2 Mice And Normotensive Bpn/3 Mice

There is considerable evidence for a role for the immune system in different animal models of hypertension. However, whether there are differences in the immune systems of Bph/2 spontaneously hypertensive mice and Bpn/3 normotensive mice is not known. To address this, we characterized the immune composition and response of male Bph/2 and Bpn/3 mice. Bph/2 mice had significantly lower body weight compared to Bpn/3 mice (Bpn: 32.0 ± 0.5 g; Bph: 23.0 ± 0.5 g) with a proportionately lower spleen weight (Bpn: 70.1 ± 9.8 mg; Bph: 48.6 ± 2.1 mg). As expected, Bph/2 mice had higher blood pressure than Bpn/3 mice. Bph/2 mice also had a higher percentage of CD4 T cells in brachial lymph nodes compared to Bpn/3 mice (Bpn: 48.0 ± 0.57, Bph: 58.6 ± 1.94) and a lower percentage of CD8 T cells (Bpn: 46.4 ± 1.07, Bph: 38.9 ± 1.84), causing an increased CD4:CD8 T cell ratio in Bph/2 mice (Bpn: 1.04 ± 0.03, Bph: 1.53 ± 0.12). This trend was also observed in inguinal lymph nodes, but not in spleens or mesenteric lymph nodes. There were also differences in the response of anti-CD3/anti-CD28-activated splenocytes. Specifically, IFNγ secretion was markedly lower in splenic T cells from Bph/2 mice compared to those from Bpn/3 mice 24 h after activation (Bpn: 4902 ± 1532 pg/ml, Bph/2: 921 ± 462 pg/ml). Similarly, IL-2 (Bpn: 3960 ± 1070 pg/ml; Bph: 1121 ± 247 pg/ml), IL-6 (Bpn: 297 ± 94 pg/ml, Bph: 64 ± 15 pg/ml) and TNFα (Bpn: 170 ± 33 pg/ml; Bph: 59 ± 15 pg/ml) induction was also lower in T cells from Bph/2 mice. Interestingly, we also found that Bpn/3 mice do not express the NK cell marker, NK1.1. Taken together, the data suggest significant differences in immune composition and response between Bph/2 and Bpn/3 mice and indicate a need to further evaluate the role of the immune system in the development of hypertension in this model.

Targeting bladder cancer: A sex sensitive perspective in mutations and outcomes.

The incidence of bladder cancer (BC) is more common in males, however, the clinical outcome for females tends to be more unfavorable, as demonstrated by a 21% increase in mortality compared to males within two years of diagnosis. While it was previously believed that the differences in outcome were solely the result of differences in sex chromosomes and hormones, it is now acknowledged that a more intricate interplay of factors is at play. By acquiring a more comprehensive understanding of these sex-specific effects, future efforts in precision medicine can be customized to an individual's biological sex. This narrative review aims to summarize our knowledge of the molecular classification of sex differences in BC by compiling the existing evidence on genetic disparities between males and females and evaluating these disparities in both noninvasive bladder cancer (NMIBC) and muscle invasive bladder cancer (MIBC). Our findings emphasize the significance of considering sex as a factor in future clinical trials and registry studies due to established differences in immune composition, molecular profiling, and genetic mutations between males and females. Further investigation into the molecular processes involved in the evasion or resistance of immune-based therapies, such as Bacillus Calmette-Guérin and other immunotherapies, is essential to identify markers of response or resistance that vary between male and female patients. This will aid in optimizing treatment and promoting equitable outcomes, particularly in NMIBC cases.

Single-cell atlas reveals different immune environments between stable and vulnerable atherosclerotic plaques.

Regardless of the degree of stenosis, vulnerable plaque is an important cause of ischemic stroke and thrombotic complications. The changes of the immune microenvironment within plaques seem to be an important factor affecting the characteristics of the plaque. However, the differences of immune microenvironment between stable and vulnerable plaques were remained unknown. In this study, RNA-sequencing was performed on superficial temporal arteries from 5 traumatic patients and plaques from 3 atherosclerotic patients to preliminary identify the key immune response processes in plaques. Mass cytometry (CyTOF) technology was used to explore differences in immune composition between 9 vulnerable plaques and 12 stable plaques. Finally, immunofluorescence technique was used to validate our findings in the previous analysis. Our results showed that more CD86+CD68+ M1 pro-inflammatory macrophages were found in vulnerable plaques, while CD4+T memory cells were mainly found in stable plaques. In addition, a CD11c+ subset of CD4+T cells with higher IFN-r secretion was found within the vulnerable plaque. In two subsets of B cells, CD19+CD20-B cells in vulnerable plaques secreted more TNF-a and IL-6, while CD19-CD20+B cells expressed more PD-1 molecules. In conclusion, our study suggested that M1-like macrophages are the major cell subset affecting plaque stability, while functional B cells may also contribute to plaque stability.

Effects of immune inflammation in head and neck squamous cell carcinoma: Tumor microenvironment, drug resistance, and clinical outcomes.

Background: Head and neck squamous cell carcinoma (HNSCC) is a malignant tumor with a very high mortality rate, and a large number of studies have confirmed the correlation between inflammation and malignant tumors and the involvement of inflammation-related regulators in the progression of HNSCC. However, a prognostic model for HNSCC based on genes involved in inflammatory factors has not been established. Methods: First, we downloaded transcriptome data and clinical information from patients with head and neck squamous cell carcinoma from TCGA and GEO (GSE41613) for data analysis, model construction, and differential gene expression analysis, respectively. Genes associated with inflammatory factors were screened from published papers and intersected with differentially expressed genes to identify differentially expressed inflammatory factor-related genes. Subgroups were then typed according to differentially expressed inflammatory factor-related genes. Univariate, LASSO and multivariate Cox regression algorithms were subsequently applied to identify prognostic genes associated with inflammatory factors and to construct prognostic prediction models. The predictive performance of the model was evaluated by Kaplan-Meier survival analysis and receiver operating characteristic curve (ROC). Subsequently, we analyzed differences in immune composition between patients in the high and low risk groups by immune infiltration. The correlation between model genes and drug sensitivity (GSDC and CTRP) was also analyzed based on the GSCALite database. Finally, we examined the expression of prognostic genes in pathological tissues, verifying that these genes can be used to predict prognosis. Results: Using univariate, LASSO, and multivariate cox regression analyses, we developed a prognostic risk model for HNSCC based on 13 genes associated with inflammatory factors (ITGA5, OLR1, CCL5, CXCL8, IL1A, SLC7A2, SCN1B, RGS16, TNFRSF9, PDE4B, NPFFR2, OSM, ROS1). Overall survival (OS) of HNSCC patients in the low-risk group was significantly better than that in the high-risk group in both the training and validation sets. By clustering, we identified three molecular subtypes of HNSCC carcinoma (C1, C2, and C3), with C1 subtype having significantly better OS than C2 and C3 subtypes. ROC analysis suggests that our model has precise predictive power for patients with HNSCC. Enrichment analysis showed that the high-risk and low-risk groups showed strong immune function differences. CIBERSORT immune infiltration score showed that 25 related and differentially expressed inflammatory factor genes were all associated with immune function. As the risk score increases, specific immune function activation decreases in tumor tissue, which is associated with poor prognosis. We also screened for susceptibility between the high-risk and low-risk groups and showed that patients in the high-risk group were more sensitive to talazoparib-1259, camptothecin-1003, vincristine-1818, Azd5991-1720, Teniposide-1809, and Nutlin-3a (-) -1047.Finally, we examined the expression of OLR1, SCN1B, and PDE4B genes in HNSCC pathological tissues and validated that these genes could be used to predict the prognosis of HNSCC. Conclusion: In this experiment, we propose a prognostic model for HNSCC based on inflammation-related factors. It is a non-invasive genomic characterization prediction method that has shown satisfactory and effective performance in predicting patient survival outcomes and treatment response. More interdisciplinary areas combining medicine and electronics will be explored in the future.

The X in seX-biased immunity and autoimmune rheumatic disease.

Sexual dimorphism in the composition and function of the human immune system has important clinical implications, as males and females differ in their susceptibility to infectious diseases, cancers, and especially systemic autoimmune rheumatic diseases. Both sex hormones and the X chromosome, which bears a number of immune-related genes, play critical roles in establishing the molecular basis for the observed sex differences in immune function and dysfunction. Here, we review our current understanding of sex differences in immune composition and function in health and disease, with a specific focus on the contribution of the X chromosome to the striking female bias of three autoimmune rheumatic diseases.

The immune landscape of common CNS malignancies: implications for immunotherapy.

Immunotherapy has enabled remarkable therapeutic responses across cancers of various lineages, albeit with some notable exceptions such as glioblastoma. Several previous misconceptions, which have impaired progress in the past, including the presence and role of the blood-brain barrier and a lack of lymphatic drainage, have been refuted. Nonetheless, a subset of patients with brain metastases but, paradoxically, not the vast majority of those with gliomas are able to respond to immune-checkpoint inhibitors. Immune profiling of samples obtained from patients with central nervous system malignancies using techniques such as mass cytometry and single-cell sequencing along with experimental data from genetically engineered mouse models have revealed fundamental differences in immune composition and immunobiology that not only explain the differences in responsiveness to these agents but also lay the foundations for immunotherapeutic strategies that are applicable to gliomas. Herein, we review the emerging data on the differences in immune cell composition, function and interactions within central nervous system tumours and provide guidance on the development of novel immunotherapies for these historically difficult-to-treat cancers.

Evolution of delayed resistance to immunotherapy in a melanoma responder.

Despite initial responses1-3, most melanoma patients develop resistance4 to immune checkpoint blockade (ICB). To understand the evolution of resistance, we studied 37 tumor samples over 9 years from a patient with metastatic melanoma with complete clinical response to ICB followed by delayed recurrence and death. Phylogenetic analysis revealed co-evolution of seven lineages with multiple convergent, but independent resistance-associated alterations. All recurrent tumors emerged from a lineage characterized by loss of chromosome 15q, with post-treatment clones acquiring additional genomic driver events. Deconvolution of bulk RNA sequencing and highly multiplexed immunofluorescence (t-CyCIF) revealed differences in immune composition among different lineages. Imaging revealed a vasculogenic mimicry phenotype in NGFRhi tumor cells with high PD-L1 expression in close proximity to immune cells. Rapid autopsy demonstrated two distinct NGFR spatial patterns with high polarity and proximity to immune cells in subcutaneous tumors versus a diffuse spatial pattern in lung tumors, suggesting different roles of this neural-crest-like program in different tumor microenvironments. Broadly, this study establishes a high-resolution map of the evolutionary dynamics of resistance to ICB, characterizes a de-differentiated neural-crest tumor population in melanoma immunotherapy resistance and describes site-specific differences in tumor-immune interactions via longitudinal analysis of a patient with melanoma with an unusual clinical course.

Abstract PS17-45: Metastatic tissues display organ specific immune infiltration archetypes; lessons from a rapid autopsy tissue collection study

Abstract Immune composition in the tumor microenvironment (TME) of patient tumors has proven to play a central role in the development of metastases and response to therapy. Evidence has suggested that the metastatic TME is immune aberrant, however difficulty in obtaining biopsies of metastatic tumors has made assessment of the immune TME difficult. Here we utilize a rapid autopsy tissue collection protocol to assess the infiltration and composition of the immune TME in numerous metastatic tissue sites, paired disease-free tissue sites, and the associated tissue draining lymph nodes. Post-mortem tissues were collected from nine metastatic breast cancer patients shortly after death through City of Hope’s “Legacy Project for Rapid Tissue Donation” Program. The average post-mortem interval (PMI) for tissue collection was 6 hours. Collected specimens include metastatic lesions and paired non-cancer samples from every cancer-involved organ, disease-free specimens from non-involved major organs, distant and tumor-draining lymph nodes (both cancer-infiltrated and disease free), as well as blood and spleens. Immediately following collection, specimens were processed into single cell suspension for flow cytometry. Over 80 immune cell phenotypes were assessed, including CD8+ and CD4+ T cell subsets, B cell subsets, natural killer (NK) cells, tumor associated macrophages (TAMs), dendritic cell subsets, and other immune cells. Tumor infiltrated tissues were found to have comparable immune cell densities and composition compared to paired disease-free tissues of the same organ type. However, immune cell densities in metastatic tissues and disease-free tissues were significantly different between organ types, with lung immune infiltration consistently being greater than liver, brain, and skin tissues. Differences in immune composition between tissue sites were also observed. Notably, liver tissues favored the presence of IL-2 producing central memory CD8+ T cells, while lung tissues favored the presence of CD8+ tissue resident memory T cells and CD16+ NK cells. Relative to disease-free lung tissues, tumor infiltrated lungs contained diminished frequencies of CD8+ tissue resident memory T cells and altered B cell and monocyte phenotypes. Increased levels of targetable immune pathways were observed, including increased PD-L1 and CTLA-4 expressing cells in skin metastases, and increased GARP+ B cells in bone marrow metastases. These data suggest that immune monitoring and trafficking of metastatic tissues site is dictated by organ type, which can be altered in composition by tumor infiltration. Further studies such as these may reveal organ-specific mechanisms of response to therapeutic interventions. Identity of organ location for tumor metastases may guide choices for immunotherapeutic interventions. Citation Format: Colt Egelston, Weihua Guo, Eliza R. Bacon, Kena Ihle, Diana Simons, Christian Avalos, JIayi Tan, Jian Ye, Lei Wang, Minhui Lim, James R. Waisman, Peter P Lee. Metastatic tissues display organ specific immune infiltration archetypes; lessons from a rapid autopsy tissue collection study [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS17-45.

Abstract 5096: The differences in immune composition observed between astrocytomas and oligodendrogliomas are due to genetics

Abstract We and others have shown that IDH-mutant astrocytomas and oligodendrogliomas share a common lineage hierarchy and a common cell of origin. However, IDH-mutant astrocytomas are infiltrated with immunosuppressive myeloid cells from the periphery to a degree not observed in IDH-mutant oligodendrogliomas. The abundance of blood-derived myeloid cells is prognostic in IDH-mutant glioma, and myeloid-derived suppressive cells are a barrier to emerging immunotherapies. Nonetheless, the cause of the differences observed between IDH-mutant subtypes is unknown. We performed single-cell RNA sequencing (scRNA-seq) and/or single-cell assay for transposon-accessible chromatin sequencing (scATAC-seq) on a cohort of 30 human primary IDH-mutant gliomas, including both astrocytomas and oligodendrogliomas. We integrated these single-cell multi-omics data to elucidate receptor-ligand interactions between tumor cells and myeloid cells, as well as the cis-regulatory grammars and associated transcription factors regulating these interactions. We validated these interactions in situ using imaging mass-cytometry on human specimens. We identified macrophage colony-stimulating factor (CSF1) and other genes responsible for myeloid cell chemotaxis, interlukin 10 (IL10), transforming growth factor beta (TGFB1) and genes responsible for promoting aberrant myeloid activation, as differentially expressed and showing differential chromatin accessibility between astrocytomas and oligodendrogliomas. The promoters of these genes were enriched for STAT3, SOX2, and SOX17/OCT4 recognition motifs, which were also differentially enriched between astrocytomas and oligodendrogliomas in our scATAC-seq data and differentially expressed in our scRNA-seq data. Since the scATAC-seq peaks in these genes correlated with known binding sites of the chromatin-modifying enzyme ATRX, and since ATRX possesses loss-of-function mutations in over %80 of IDH-mutant astrocytomas but almost never in oligodendrogliomas, we hypothesize that the observed differences in myeloid abundance are driven by a loss of ATRX binding in key chemotactic and immunosuppressive cytokines and their upstream transcription factors. To test this hypothesis, we knocked out ATRX in a a syngeneic, immunocompetent glioma mouse model using CRISPR and profiled the resulting tumors via a battery of genomics and protein-level assays. We conclude that the differences in immune composition between IDH-mutant glioma subtypes are due to genetics. In particular, ATRX loss of function in IDH-mutant astrocytoma leads to chromatin changes in STAT3 and downstream effector cytokines, resulting in a greater influx of immunosuppressive myeloid cells. Citation Format: Lin Wang, Husam Babikir, Karin Shamardani, Francisca Catalan, Aaron Diaz. The differences in immune composition observed between astrocytomas and oligodendrogliomas are due to genetics [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5096.

Organ specificity dictates tumor immune infiltration and composition in metastatic breast cancer; lessons from a rapid autopsy tissue collection study.

1032 Background: Immune composition in the tumor microenvironment (TME) of patient tumors has proven to play a central role in the propensity of tumors to metastasize and respond to therapy. Evidence has suggested that the metastatic TME is immune aberrant, however limited sample size and numbers has made assessment of the immune TME in the development of multi-organ metastases difficult. Here we utilize a rapid autopsy tissue collection protocol to assess the infiltration and composition of the immune TME in numerous metastatic tissue sites, paired disease-free tissue sites, and the associated tissue draining lymph nodes. Methods: Post-mortem tissues were collected from six metastatic breast cancer patients shortly after death through City of Hope’s “Legacy Project for Rapid Tissue Donation” Program. The average post mortem interval (PMI) for tissue collection was 6 hours. Collected specimens include metastatic lesions and paired non-cancer samples from every cancer-involved organ, disease-free specimens from non-involved major organs, distant and tumor-draining lymph nodes (both cancer-infiltrated and disease free), as well as blood. Immediately following collection, specimens were processed into single cell suspension for flow cytometry. Over 80 immune cell phenotypes were assessed, including CD8+ and CD4+ T cell subsets, B cell subsets, natural killer (NK) cells, tumor associated macrophages (TAMs), dendritic cell subsets, and other cells. Results: Tumor infiltrated tissues were found to have comparable immune cell densities and composition compared to paired disease-free tissues of the same organ type. However, immune cell densities in metastatic tissues and disease-free tissues were significantly different between organ types, with lung immune infiltration consistently being greater than liver tissues. Differences in immune composition between tissue sites were also observed. Notably, liver tissues favored the presence of central memory CD8+ T cells, while lung tissues favored the presence of CD8+ tissue resident memory T cells. Relative to disease-free lung tissues, tumor infiltrated lungs contained diminished frequencies of CD8+ tissue resident memory T cells and altered B cell phenotypes. Conclusions: These data suggest that immune monitoring and trafficking of metastatic tissues site is dictated by organ type, which can be altered in composition by tumor infiltration. Further studies such as these may reveal organ-specific mechanisms of response to therapeutic interventions.

Examining the Immune Composition of the Intestinal Mucosa by Mass Cytometry

Abstract BACKGROUND The inflammatory bowel diseases (IBD) can affect various segments of the gastrointestinal tract, and the biology behind regional differences in disease remains poorly understood, in part due to a lack of understanding of regional variation of the particular cell subsets present. By utilizing mass cytometry, our objective was to identify regional differences in the immune composition of the digestive tract in healthy individuals. METHOD Intestinal pinch biopsies were obtained from the following sites of healthy individuals (n=10); duodenum, terminal ileum, ascending colon, transverse colon, descending colon, and rectum. Immune cells were isolated from biopsies and stained with a panel of 37 markers for analysis using mass cytometry. RESULTS Marked variations in immune cell composition were observed along the digestive tract, in particular T and B lymphocytes. Increased abundance of T lymphocytes (>30% of total CD45+ cells) was identified in the duodenum and terminal ileum, whereas B lymphocytes became more abundant (>30% of total CD45+ cells) towards the distal end of the tract. In addition, intestinal homing molecules displayed variation at each site; integrin α4β7, a target for IBD therapeutic intervention, was increasingly expressed in the terminal ileum and ascending colon, compared with the descending colon (Mean 40% vs 25% abundance, n=10, p<0.01). CONCLUSIONS Mass cytometry has the ability to identify and characterize numerous distinct immune subsets throughout the intestinal mucosa, allowing a deep understanding of regional differences in immune composition. This understanding may shed light on the pathogenesis of IBD and allow standardization of intestinal sampling during the evaluation of therapeutics.