Identifying a potential biomarker for anti-PD-1 immunotherapy in patients with advanced stage, surgically-resectable endometrial cancer.

5590 Background: The FDA approval of pembrolizumab for patients with MSI-H or dMMR tumors has led to the treatment of a select cohort of endometrial cancer (EC) patients. We sought to ascertain tumor immune modulatory effects in the front-line setting for advanced stage III/IV EC patients regardless of MSI-H or dMMR. The primary objective was to determine the safety of preoperative and maintenance pembrolizumab. The secondary objective was to examine pembrolizumab-induced changes in peripheral immune effector phenotype in order to identify potential biomarkers of clinical response. Methods: In an open label, single-arm Phase I trial, 8 EC patients were treated with 2 doses of preoperative pembrolizumab IV prior to surgery followed by chemotherapy and 4 doses of pembrolizumab IV. As an initial study, pre- and post-treatment (on the day of surgery) peripheral blood was collected from 3 patients as well as a healthy control and processed for high-dimensional single-cell mass cytometry (CyTOF) using an optimized antibody panel. Results: Six of 8 patients completed the treatment. One patient had rapid cancer progression and another had an exacerbation of comorbidities. Peripheral blood from 3 patients with pathological response were then immunoprofiled using CyTOF. Data analysis revealed that the frequencies of CD8+ T cells, B cells and CD56hiCD16- NK cells were lower, whereas the frequency of CD14+CD16-HLA-DRhi classical monocytes was higher in the cancer patients compared to controls. Cancer patients had lower frequencies of circulating CD4+ and CD8+ naïve T cells but higher frequencies of effector CD8+ and CD4+ T cells. Notably, the median expression of Granzyme B in CD8+ and CD4+ T cells was higher and median expression of signal regulatory protein (SIRP), CD172a-b on monocytes was lower for cancer patients compared to control. The frequencies of NK and myeloid cells expressing CD137(4-1BB), PD-1+NK cells, and PD-L1+DCs were greater in post-compared to pre-pembrolizumab. Conclusions: This is the first trial to evaluate the use of neoadjuvant pembrolizumab in advanced stage EC patients. Here, we present peripheral immune correlative data and show an increase in markers of activation in patients with pathologic responses to pembrolizumab. Additional data from this ongoing study will help us to identify candidate predictive biomarkers. Clinical trial information: NCT02630823.

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Proliferation of PD-1+ CD8 T cells in peripheral blood after PD-1–targeted therapy in lung cancer patients

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1705327114 ◽

2017 ◽

Vol 114 (19) ◽

pp. 4993-4998 ◽

Cited By ~ 229

Author(s):

Alice O. Kamphorst ◽

Rathi N. Pillai ◽

Shu Yang ◽

Tahseen H. Nasti ◽

Rama S. Akondy ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cancer Patients ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Cd8 T Cell ◽

Blood Analysis ◽

Advanced Stage ◽

Nsclc Patients ◽

Peripheral Blood Analysis

Exhausted T cells in chronic infections and cancer have sustained expression of the inhibitory receptor programmed cell death 1 (PD-1). Therapies that block the PD-1 pathway have shown promising clinical results in a significant number of advanced-stage cancer patients. Nonetheless, a better understanding of the immunological responses induced by PD-1 blockade in cancer patients is lacking. Identification of predictive biomarkers is a priority in the field, but whether peripheral blood analysis can provide biomarkers to monitor or predict patients’ responses to treatment remains to be resolved. In this study, we analyzed longitudinal blood samples from advanced stage non–small cell lung cancer (NSCLC) patients (n = 29) receiving PD-1–targeted therapies. We detected an increase in Ki-67+ PD-1+ CD8 T cells following therapy in ∼70% of patients, and most responses were induced after the first or second treatment cycle. This T-cell activation was not indiscriminate because we observed only minimal effects on EBV-specific CD8 T cells, suggesting that responding cells may be tumor specific. These proliferating CD8 T cells had an effector-like phenotype (HLA-DR+, CD38+, Bcl-2lo), expressed costimulatory molecules (CD28, CD27, ICOS), and had high levels of PD-1 and coexpression of CTLA-4. We found that 70% of patients with disease progression had either a delayed or absent PD-1+ CD8 T-cell response, whereas 80% of patients with clinical benefit exhibited PD-1+ CD8 T-cell responses within 4 wk of treatment initiation. Our results suggest that peripheral blood analysis may provide valuable insights into NSCLC patients’ responses to PD-1–targeted therapies.

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Peripheral blood gamma-delta T cells in advanced-stage cancer patients

Advances in Therapy ◽

10.1007/s12325-008-0032-z ◽

2008 ◽

Vol 25 (3) ◽

pp. 218-224 ◽

Cited By ~ 5

Author(s):

Oguz Bilgi ◽

Bülent Karagoz ◽

Orhan Turken ◽

E. Gokhan Kandemir ◽

Ahmet Ozturk ◽

...

Keyword(s):

T Cells ◽

Cancer Patients ◽

Peripheral Blood ◽

Gamma Delta T Cells ◽

Advanced Stage ◽

Gamma Delta ◽

Advanced Stage Cancer

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CD8+CD28− cells and CD4+CD25+ regulatory T cells in the peripheral blood of advanced stage lung cancer patients

Medical Oncology ◽

10.1007/s12032-008-9165-9 ◽

2009 ◽

Vol 27 (1) ◽

pp. 29-33 ◽

Cited By ~ 53

Author(s):

Bülent Karagöz ◽

Oğuz Bilgi ◽

Mahmut Gümüs ◽

Alev Akyol Erikçi ◽

Özkan Sayan ◽

...

Keyword(s):

Lung Cancer ◽

T Cells ◽

Regulatory T Cells ◽

Cancer Patients ◽

Peripheral Blood ◽

Advanced Stage ◽

Lung Cancer Patients ◽

Advanced Stage Lung Cancer ◽

Stage Lung Cancer

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CHAPTER 9: Immunology of TBEV-Infections

Tick-borne encephalitis - The Book ◽

10.33442/26613980_9-3 ◽

2020 ◽

Keyword(s):

Central Nervous System ◽

T Cells ◽

Cerebrospinal Fluid ◽

Nervous System ◽

Nk Cells ◽

Peripheral Blood ◽

Sample Collection ◽

Tick Borne Encephalitis ◽

Viral Infectious Disease ◽

The Central Nervous System

Tick-borne encephalitis (TBE) is a viral infectious disease of the central nervous system caused by the tick-borne encephalitis virus (TBEV). TBE is usually a biphasic disease and in humans the virus can only be detected during the first (unspecific) phase of the disease. Pathogenesis of TBE is not well understood, but both direct viral effects and immune-mediated tissue damage of the central nervous system may contribute to the natural course of TBE. The effect of TBEV on the innate immune system has mainly been studied in vitro and in mouse models. Characterization of human immune responses to TBEV is primarily conducted in peripheral blood and cerebrospinal fluid, due to the inaccessibility of brain tissue for sample collection. Natural killer (NK) cells and T cells are activated during the second (meningo-encephalitic) phase of TBE. The potential involvement of other cell types has not been examined to date. Immune cells from peripheral blood, in particular neutrophils, T cells, B cells and NK cells, infiltrate into the cerebrospinal fluid of TBE patients.

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186 Distinct immune signatures predicting clinical response to PD-1 blockade therapy in gynecological cancers revealed by high-dimensional immune profiling

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0186 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A200-A200

Author(s):

Yuki Muroyama ◽

Sasikanth Manne ◽

Alexandar Huang ◽

Divij Mathew ◽

...

Keyword(s):

Ovarian Cancer ◽

T Cells ◽

Endometrial Cancer ◽

T Cell ◽

Clinical Response ◽

Cancer Patients ◽

T Cell Response ◽

High Dimensional ◽

Uterine Endometrial Cancer ◽

Immune Profiling

BackgroundAlthough immune checkpoint blockade revolutionized cancer therapy, response rates have been mixed in gynecological malignancies. While uterine endometrial cancer with high microsatellite instability (MSIHI) and high tumor mutational burden (TMB) respond robustly to checkpoint blockade, high-grade serous ovarian cancer (HGSOC) with low TMB respond modestly. Currently, there has been no known immune signature or T cell phenotype that predicts clinical response in gynecological tumors.MethodsTo dissect the immune landscape and T cell phenotypes in gynecological cancer patients receiving PD-1 blockade, we used high-dimensional cytometry (flow cytometry and mass cytometry (CyTOF)). We performed longitudinal deep immune profiling of PBMC from patients with recurrent uterine endometrial cancer receiving single-arm nivolumab, and HSGOC patients receiving neoadjuvant nivolumab plus platinum-based chemotherapy prior to debulking surgery.ResultsChemotherapy-resistant MSI-H uterine cancer patients treated with nivolumab had a proliferative T cell response 2–4 weeks post PD-1 blockade, consistent with responses seen in high TMB melanoma and lung cancer. The responding Ki67+ CD8 T cell population was largely CD45RAloCD27hi or CD45RAloCD27lo and highly expressed PD1, CTLA-4, and CD39, consistent with the phenotype of exhausted T cells (TEX). These exhausted-like cells are enriched in responders, whereas early expansion Tregs are enriched in non-responders. Unlike patients with uterine endometrial cancer, patients with TMBlo ovarian cancer did not have a clear proliferative CD8 T cell response after neoadjuvant nivolumab plus chemotherapy treatment, suggesting systemic immune suppression. At baseline, ovarian cancer without recurrence have more terminally differentiated effector-like CD8 T cells, and patients with recurrence have more naive-like cells. Thus, both high and low TMB gynecological tumors have distinct immune landscapes associated with clinical response. Additionally, in MSI-H uterine endometrial cancer patients, the length of time between the prior chemotherapy and the initiation of immunotherapy was negatively correlated with T cell reinvigoration post immunotherapy and clinical response. This suggests the importance of optimize therapeutic timing to maximize the therapeutic efficacy when combining immunotherapy and chemotherapy.ConclusionsCollectively, our immune profiling revealed the distinct immune signatures associated with clinical response to PD-1 blockade in gynecological cancers. Our results also suggest that TMBhi inflamed versus TMBlo cold tumor microenvironment, and timing of chemo/immunotherapy could impact differentiation and functions of T cells.Ethics ApprovalThe study was approved by MSKCC Ethics Board, approval number 17–180 and 17–182.

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Erratum to: Content of HLA-G+ T Cells in the Peripheral Blood from Healthy Women and Breast Cancer Patients

Bulletin of Experimental Biology and Medicine ◽

10.1007/s10517-016-3224-6 ◽

2016 ◽

Vol 160 (4) ◽

pp. 592-592

Author(s):

E. O. Ostapchuk ◽

Yu. V. Perfil’eva ◽

Sh. Zh. Talaeva ◽

N. A. Omarbaeva ◽

N. N. Belyaev

Keyword(s):

Breast Cancer ◽

T Cells ◽

Cancer Patients ◽

Peripheral Blood ◽

Breast Cancer Patients ◽

Healthy Women

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Chapter 9: Immunology of TBEV-Infection

Tick-borne encephalitis - The Book ◽

10.33442/26613980_9-4 ◽

2021 ◽

Author(s):

Sara Gredmark-Russ ◽

Renata Varnaite

Keyword(s):

Central Nervous System ◽

T Cells ◽

Cerebrospinal Fluid ◽

Nervous System ◽

Nk Cells ◽

Peripheral Blood ◽

Sample Collection ◽

Tick Borne Encephalitis ◽

Viral Infectious Disease ◽

The Central Nervous System

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151 Potentiating the Large-Scale Expansion and Engineering of Peripheral Blood-Derived CAR NK Cells for Off-the-Shelf Application

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2021-sitc2021.151 ◽

2021 ◽

Vol 9 (Suppl 3) ◽

pp. A159-A159

Author(s):

Michael Whang ◽

Ming-Hong Xie ◽

Kate Jamboretz ◽

Hadia Lemar ◽

Chao Guo ◽

...

Keyword(s):

T Cells ◽

Cell Therapy ◽

Nk Cells ◽

Peripheral Blood ◽

Nk Cell ◽

Adoptive Cell Therapy ◽

Therapeutic Window ◽

Target Cells ◽

Healthy Donors ◽

Custom Made

BackgroundPeripheral blood natural killer (NK) cells are mature cytotoxic innate lymphocytes possessing an inherent capacity for tumor cell killing, thus making them attractive candidates for adoptive cell therapy. These NK cells are also amenable to CRISPR and chimeric antigen receptor (CAR) genomic engineering for enhanced functions. Moreover, NK cells possess an inherent capacity for off-the-shelf therapy since they are not known to cause graft-versus-host disease, unlike T cells. Presently, approved CAR cell therapy is custom-made from each patient‘s own T cells, a process that can limit patient pool, narrow therapeutic window, and contribute to product variability. In this study, we investigate whether peripheral blood NK cells from a selected donor can be edited, engineered, and expanded sufficiently for off-the-shelf use in a wide patient population.MethodsUsing the CRISPR/Cas9 system, we knocked out CISH expression in isolated peripheral blood NK cells from 3 healthy donors. Subsequently, we expanded edited NK cells by using IL-2 and sequential stimulations using NKSTIM, a modified K562 stimulatory cell line expressing membrane-bound form of IL-15 (mbIL-15) and 4-1BBL. IL-12 and IL-18 were added twice during expansion to drive memory-like NK cell differentiation. We transduced the expanded NK cells to express engineered CD19-targeted CAR and mbIL-15 during an interval between the first and second NKSTIM pulses. We assessed NK cell cytotoxicity against Nalm6 target cells by IncuCyte.ResultsIsolated peripheral blood NK cells from 3 healthy donors were successfully edited using CRISPR/Cas9, engineered to express high levels of CAR, extensively expanded using a series of NKSTIM pulses in the presence of IL-2, and differentiated into memory-like NK cells using IL-12 and IL-18. Interestingly, NK cells from the 3 donors exhibited distinct outcomes. NK cells from one donor reached a peak expansion limit of approximately 7-million-fold before undergoing contraction whereas NK cells from two donors continued to expand over the length of the study surpassing 100-million-fold expansion, without appearing to have reached a terminal expansion limit. At the end of the study, NK cells from one donor exceeded 1-billion-fold expansion and maintained 88% cytolytic activity compared to Nkarta’s standard process control in a 72-hour IncuCyte assay.ConclusionsIn this study, we demonstrate that healthy donor-derived peripheral blood NK cells are capable of expanding over billion-fold while maintaining potency. These results provide a rationale for the development of off-the-shelf CAR NK cell therapies using NK cells from donors selected to provide optimal product characteristics.Ethics ApprovalHuman samples were collected with written informed consent by an approved vendor.

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