bluuebunny/crossref_metadata_2025 huggingface dataset:
Potential elimination of multi-drug resistant cancer cells by targeted non-engrafting intentionally mismatched activated donor lymphocytes.
Authors: Slavin Shimon
Published: May 2019

e14007

Background: Immunotherapy by targeted autologous T cells, TIL or CAR-T, are considered most effective modalities for treatment of otherwise resistant cancer, yet cure is rarely achieved. In contrast, cure of resistant disease by donor lymphocyte infusion (DLI) following allogeneic stem cell transplantation suggests that alloreactive lymphocytes represent a most effective approach for elimination of resistant malignant cells. The first patient relapsing following supra-lethal chemoradiotherapy is alive & well > 34 years. Using murine B cell leukemia (BCL1) and metastatic breast cancer (4T1) we proved that non-engrafting intentionally mismatched lymphocytes activated with IL-2 that were consistently rejected cured otherwise lethal cancer with no graft-vs-host disease (GVHD). Methods: We first applied graft-vs-tumor effects by non-engrafting Intentionally Mismatched Activated Killers (IMAK) following mild immunosuppressive conditioning to maximize homeostatic proliferation of activated T & NK cells. IMAK was first applied inpatients with multiple myeloma and lymphoma treated with donor lymphocytes pre-activated in vitro prior to infusion and for 5 days following infusion withlow dose IL-2.We next studied feasibility of Allogeneic Targeted Activated Cancer Killer cells (ATACK) using IL-2 activated haploidentical donor lymphocytes including T & NK cells targeted against antigens over-expressed on the surface of malignant cells: Her-2/neu, EGFR, VEGF and CD20 using commercially available Herceptin, Erbitux, Avastin or Rituximab, respectivelyin a pilot study involving 16 patients with advanced metastatic solid tumors. Results: Using IMAK donor lymphocytes were uniformly rejected and no patient developed GVHD. Disease-free survival > 20 years was documented in our first myeloma patient with plasmacytomas and in another relapsing after autologous SCTand in 2 patients with non-Hodgkin lymphoma. Longterm progression-free survival was reported in 5 of 31 patients with resistant solid tumors. Following ATACK transient mild toxicity was manageable in outpatient clinic and no GVHD developed. Conclusions: Using ATACK against minimal residual disease accomplishable by most cancer patients following conventional 1st line treatment may result in cure of otherwise resistant cancer while avoiding GVHD.
Development of a novel dendritic cell-based immunotherapy targeting cancer stem cells.
Authors: Calmeiro João , Carrascal Mylène , Mendes Luís , Duarte Iola F. , Gomes Célia , Serra João , Falcão Amilcar , Cruz Maria Teresa , Neves Bruno Miguel
Published: May 2019

e14009

Background: Dendritic cells (DCs) are one of the central tools in cellular anti-tumor immunotherapy, being characterized by their capacity for acquiring and processing antigens and ability to produce strong antitumor immune responses. The production of clinical grade ex-vivo monocyte-derived DCs (Mo-DCs) is the most frequent approach for antitumor vaccines production. Recently, therapeutic resistance to radio/chemotherapy and disease recurrence was shown to be in part due to a small cancer stem cell (CSCs) population present in tumors. Methods: Here, we aim to target and eradicate CSCs by developing a novel DC-based immunotherapy vaccine for pancreatic and non-small cells lung cancer (NSCLC), comparing the loading of CSCs vs. classical tumor lysates. Results: CSCs from PANC-1 (pancreatic cancer) and A549 (NSCLC) cell lines were successfully isolated and characterized, overexpressing stem-like markers: NANOG, OCT4, SOX2 and CD133. CSCs resistance to Gemcitabine was also assessed. Before comparing the 2 types of vaccine loading, we also analyzed the impact of 3 GMP free-serum culture media on the phenotype and functional abilities of Mo-DCs. DCs cultured in X-VIVO 15 and AIM-V media show enhanced production of IL-12 and are able to induce a superior stimulation of T cells, mainly CTLs and Th1 subsets. By contrast, DCs cultured in DendriMACS are more prone to induce Treg polarization. Conclusions: Overall, our data demonstrate that blood monocytic precursors present considerable plasticity allowing a tailored differentiation of DCs just by changing the nutritive support. We also highlight the need of critically defining the culture medium to be used in DC cancer immunotherapy in order to attain desired cell characteristics and by consequent robust responses. Finally, our preliminary results indicate that loading DCs with CSCs antigens may be an effective strategy to target and destroy this resilient cancer cell population.
A phase Ib clinical trial on intratumoral administration of autologous CD1c (BDCA-1)+ myeloid dendritic cells (myDC) in combination with ipilimumab (IPI) and avelumab (AVE) plus intravenous low-dose nivolumab (NIVO) in patients with advanced solid tumors.
Authors: Schwarze Julia Katharina , Awada Gil , Cras Louise , Forsyth Ramses , Van Riet Ivan , Neyns Bart
Published: May 2019

e14012

Background: Intratumoral (IT) myDC play a pivotal role in initiating antitumor immune responses and "re-licensing” of antitumor cytotoxic T-lymphocytes within the tumor microenvironment. IT injection of anti-PD-L1 IgG1 mAb AVE and anti-CTLA-4 IgG1 mAb IPI may reduce the number of regulatory T cells and lyse PD-L1+ tumor cells, thereby releasing tumor antigens that can be captured and processed by IT co-administered CD1c (BDCA-1)+ myDC, reinvigorating the cancer immunity cycle. Methods: Patients (pts) with advanced solid tumors who failed standard therapy were eligible for IT injections of ≥1 non-visceral metastasis with IPI (max total dose of 10 mg) and AVE (max total dose of 40 mg) plus IV NIVO (10 mg) on day 1 followed by IT injection of autologous, non-substantially manipulated CD1c (BDCA-1)+ myDC on day 2. Administration of AVE, IPI, and NIVO was repeated every 14 days thereafter. Primary endpoints were safety and feasibility. Repetitive FNA cytology/IHC of treated lesions was performed. Results: In this ongoing trial, 6 pts (3x melanoma, 1x epithelial ovarian carcinoma, 2x triple negative breast carcinoma) were treated with IT injection of a median of 27,2x106 (range 10-43x106) CD1c (BDCA-1)+ myDC and a median of 5 (range 2-10) study drug administrations. At time of this analysis 3 pts are evaluable for response: an ongoing PR ( > 8 months) was documented in a melanoma pt who previously progressed on PD-1 and CTLA-4 inhibitors. In 2 other melanoma pts regression of the injected metastases coincided with progression of non-injected metastases. Adverse events consisted of transient grade(G)2 local pain at injection site in 2 pts, G1 pruritus in 2 pts, G2 pneumonitis in 1 pt, G1 rash in 1 pt, and pruritus and redness of the skin overlaying the injected lesion in 1 pt. Analysis of cytology/IHC results is ongoing. Conclusions: IT injection of autologous CD1c (BDCA-1)+ myDC with IT co-injection of AVE and IPI plus IV low-dose NIVO is feasible and tolerable and resulted in encouraging early signs of anti-tumor activity in injected as well as non-injected lesions. Clinical trial information: NCT03707808.
Long-term survival in patients (pts) with locally advanced or disseminated malignant melanoma (MM) or renal cell carcinoma (RCC), treated with tag-7 gene-modified (GMV) tumor cells: A phase II trial.
Authors: Novik Alexey Viktorovich , Danilova Anna Borisovna , Baldueva Irina Alexandrovna , Sluzhev Maksim Ivanovich , Nehaeva Tatyana Leonidovna , Larin Sergei , Protsenko Svetlana , Semenova Anna , Danilov Aleksei Olegovich , Moiseyenko Vladimir , Georgiev Georgii Pavlovich
Published: May 2019

e14022

Background: Immunotherapy may produce long lasting effects on survival.Magnitude of efficacy may be depended from immune factors. We evaluated the overall survival (OS) in pts treated with GMV with biomarker analysis of immunosuppressive factors (ISF) production by their tumor cells. Methods: Since 2001 to 2014 80 pts received GMV: 68 with MM and 12 pts with RCC. Treatment was given in the metastatic setting in 61 pts (MM-51, RCC – 10) and in the adjuvant (after complete cytoreduction) setting in 19 pts (MM-17, RCC-2). Stage 3 was in 26 (33%) pts, stage 4 in 54 (67%). Pt’s tumor samples were transferred to culture, transfected with TAG7and inactivated by radiation. Produced product was injected SC every 3 weeks until progression or 2y of therapy. ISF (MICA, TGF-β1, IL-10 and VEGF) were measured in the supernatants of the tumor cell cultures and used as predictive factors. Results: The 5-yr OS in intention to treat population was 25.1%. There was no differences in MM and RCC OS (Log-rank p = 0.44). Median OS in metastatic setting was 0,7 y, in adjuvant – 3,1 y. Classification trees were built on the basis of ISF production. The median OS was 6.6 y in favorable prognosis (FP) group (MICA level ≤582 pg/ml, n = 15), 4.6 mo in unfavorable (UF) group (MICA level > 582 pg/ml, n = 12) (p < 0.0001). No significant differences were found between classification trees based on the other ISF (TGF-β1, IL-10 and VEGF). In stage 3-4 MM pts with FP median OS was 2,3 y with 31% pts alive in 10y, in UF group – 0,4 y; log-rank p = 1,94E-5. No FP pts received modern immunotherapy. Conclusions: GMV shows high results in carefully selected pts with low ISF production. Method should be further investigated in pts with FP. [Table: see text]
Stochastic cancer-immune coevolution: Implications for cancer incidence and immunotherapeutic efficacy.
Authors: George Jason T , Levine Herbert , Molldrem Jeffrey J. , Garber Haven
Published: May 2019

e14023

Background: Despite recent progress, robust treatment strategies that lead to durable remission are still lacking for many cancer types. This disease is difficult to treat owing in part to the complexity introduced by a heterogeneous population of cancer cells capable of evolving mechanisms of resistance to traditional therapy. Nonetheless, the discovery and continued optimization of T-cell immunotherapy has revolutionized the treatment of many cancers. This treatment strategy stands out from other approaches in its unique ability to co-evolve alongside an evading tumor. While promising, such therapies are also complex. For example, allogeneic stem cell transplantation leverages a donor-derived T-cell repertoire to treat patients with refractory hematologic malignancies and relies upon a delicate balance between desirable anti-tumor effects and potentially life-threatening graft-versus-host-disease. Currently, the decision to utilize this therapy and others like it is largely influenced by prior empirical evidence. Thus, there is great need for quantitative models of the cancer-immune interaction to generate testable predictions of treatment outcome, which could then be validated prior to T-cell immunotherapy administration. Methods: We develop a foundational mathematical model to investigate the properties of stochastic tumor-immune co-evolution using applied stochastic process theory and probabilistic analysis. We use this model to predict the effects of reduced immunity, T-cell diversity, and thymic turnover rates on cancer incidence, and compare model simulations to cancer evolutionary data. Results: We predict that changes in T-cell diversity, and to a lesser degree thymic turnover, increase the chance of tumor progression. When applied to experimental data, we demonstrate that the observations are consistent with co-evolution between an indolent cancer population and the adaptive immune system prior to clinical disease. Conclusions: Our results provide a fundamental framework for analyzing the interaction dynamics of an evolving threat like cancer and the adaptive immune system in order to better understand and predict immunotherapeutic efficacy.
Treatment with tumor-infiltrating lymphocytes in the changing treatment landscape of metastatic melanoma.
Authors: Borch Troels , Andersen Rikke , Ellebaek Eva , Met Özcan , Donia Marco , Svane Inge Marie
Published: May 2019

e14024

Background: Adoptive cell therapy using tumor-infiltrating lymphocytes (TIL) has been shown to induce durable complete responses in patients with metastatic melanoma (MM) who are anti-PD-1 naïve. Whether progression on or after anti-PD-1 therapy affects the outcome of TIL therapy given as a subsequent treatment line is largely unknown. To elucidate this, we analyzed updated clinical data covering a decade of TIL trials carried out in Denmark. Methods: Data from three clinical trials were pooled and data from 55 treated patients were available (ClinicalTrials.gov Identifiers: NCT00937625 (n = 31), NCT02379195 (n = 12) and NCT02354690 (n = 12)). Survival curves were computed according to the Kaplan-Meier method. Clinical response rate (RR) was evaluated according to RECIST criteria. Results: Median overall survival in the pooled cohort was 15.9 months and progression-free survival (PFS) was 3.7 months. Six patients achieved a complete response (11%), of which four are ongoing. Fourteen patients achieved a partial response (26%), of which three are ongoing. Median overall survival of responders was not reached with a median follow-up time of 40 months. RR was not statistically different depending on prior anti-PD-1 therapy (42% no prior anti-PD-1 therapy vs. 32% with prior anti-PD-1 therapy). However, there was a trend towards a shorter duration of partial responses in patients previously treated with both anti-CTLA-4 monotherapy and anti-PD-1 monotherapy, compared to patients previously treated with anti-CTLA-4 but not anti-PD-1 (P = 0.06). The two groups were balanced in respect to number of prior treatment lines. Conclusions: After progression on anti-PD-1 therapy, partial responses following TIL therapy might be shorter, but durable complete responses can be induced despite progression on prior anti-PD-1 therapy. Thus, TIL therapy remains an important treatment strategy in MM. Clinical trial information: NCT00937625, NCT02379195, NCT02354690.
Safety and efficacy of a phase I/IIa trial (NCT03066947) of a modified whole tumor cell targeted immunotherapy in patients with advanced breast cancer.
Authors: Williams William , Holmes Jarrod P. , Bhattacharya Saveri , Calfa Carmen , Dakhil Shaker R. , Lukas Jason Jerome , Tan-Chiu Elizabeth , Adams Daniel , Peoples George , Lacher Markus , Wiseman Charles L.
Published: May 2019

e14026

Background: SV-BR-1-GM is a GM-CSF transfected breast cancer cell line which expresses HLA class I & II antigens and has functional antigen-presenting cell activity. Prior studies suggest that partial matching of the HLA type of the patient with SV-BR-1-GM may be predictive of tumor regression. Methods: Subjects received low-dose cyclophosphamide 2-3d prior to ID injection of irradiated SV-BR-1-GM (20 million cells divided into 4 sites) and interferon-α into the inoculation sites ~2 & 4 days subsequently. Cycles were q2 weeks x 3 then q mo. Results: A total of 30 patients were screened and 23 inoculated (Table). The patients were heavily pretreated with a median of 4 prior chemo/biological therapy regimens. There were no serious or unexpected adverse events. Local injection-site irritation was the most common toxicity. Objective tumor regression was seen in 3 patients, all of whom matched SV-BR-1-GM at least at one HLA locus: one patient with regression or clearing of 20 lung metastases; one with reduction in cutaneous involvement of the breast from 80% to 30% and one with regression of a breast lesion. Another 3 patients had decreases in circulating cancer-associated macrophage-like cells (CAMLs), which has been shown to correlate with tumor stage. They also all matched at least at one HLA allele. Circulating tumor cells and circulating epithelial cells were present in low numbers and tended to parallel trends in CAMLs which were present in larger numbers. CAMLs in 21/23 patients stained positive for PD-L1. Patients with tumor regression had robust DTH responses to SV-BR-1-GM. Conclusions: SV-BR-1-GM in this regimen appears to be safe and well-tolerated and is associated with objective regression of metastatic breast cancer and/or with decreases in circulating cancer-associated cells in 6/23 (26%) or patients. HLA matching may be a predictor of response. Clinical trial information: NCT03066947. [Table: see text]
Prognostic interaction of smoking history with circulating regulatory T cells in advanced non-small cell lung cancer.
Authors: Liu Chao , Sun Yi , Yue Jinbo , Yu Jinming
Published: May 2019

e14030

Background: Smokers with non-small cell lung cancer (NSCLC) respond better to the inhibition of programmed cell death-1 (PD-1) with pembrolizumab than non-smokers do, which may be accounted for by the fact that smoking induces higher neoantigens and increased immunogenicity. Our investigation sought to highlight the prognostic interaction of smoking history with circulating regulatory T cells (Treg), CD3+, CD4+, and CD8+ T cells in advanced NSCLC. Methods: Using ﬂow cytometry, we assessed levels of circulating Treg cells, CD8+ T cells, CD4+ T cells, and CD3+ T cells in 106 eligible patients of 257 overall patients with advanced NSCLC. Results: Patients with high circulating Treg had shorter overall survival (OS) (12.5 vs. 27.5 months, P = 0.010) and progression-free survival (PFS) (7.1 vs. 16.9 months, P = 0.014) compared to patients with low Treg among non-smokers, but not among smokers (P = 0.211 for OS; P = 0.271 for PFS). Multivariate analysis showed Treg as an independent risk factor for outcomes of non-smoking patients [hazard ratio (HR): 2.71, P = 0.045 for OS; HR: 2.38, P = 0.047 for PFS], but not of smokers. Moreover, higher CD4+ T and lower CD8+ T cell counts, CD8/CD4 ratio, and CD8/Treg ratio were more abundant in non-smokers than smokers (all P < 0.05). Conclusions: Smoking impacts the prognostic value of circulating Treg in advanced NSCLC. The distribution of circulating immune cells in non-smokers and smokers was different. Assessing the effect of cigarette smoking on circulating immune cells could help individualized immunotherapy for advanced NSCLC.
Effect of radiotherapy on the residual circulating epithelial tumor cells (CETCs) and their programmed cell death ligand 1 (PD-L1) status in breast cancer patients.
Authors: Schott Dorothea , Pizon Monika , Pachmann Katharina , Pachmann Ulrich A. , Maeurer Matthias
Published: May 2019

e14034

Background: Preclinical studies have suggested that radiotherapy (RT) followed by immunotherapy could be a promising strategy for synergistic enhancement of treatment efficacy. RT induces immunogenic changes in cancer cells which can lead to adaptive up-regulation of PD-L1 expression. Moreover, radiotherapy reduces not only local recurrence but also improves overall survival by preventing distant metastases, which indicates its influence on the remaining occult tumor, such as circulating tumor cells. For this reason the aim of this study was to determine the PD-L1 status on CETCs before and during RT. Methods: CETCs were determined in 54 primary breast cancer patients in early and locally advanced stages. The number of CETCs and their expression of PDL-L1 were investigated using the maintrac method. The fraction of PD-L1 positive CETCs was assessed prior to (baseline), 3 and 6 weeks after the start of radiotherapy. Results: We found a correlation between the PD-L1 status of CETCs and aggressiveness of cancer disease prior to radiotherapy. Patients with positive lymph node status presented more PD-L1 positive CETCs compared to patients with negative lymph nodes (mean 73% vs. 50%, p < 0.01). Moreover, the HER2 expression of the primary tumor was also positively correlated with PD-L1 expression. Lower numbers of CETCs were detected in patients who had not received chemotherapy or adjuvant chemotherapy before RT than in those who received neoadjuvant chemotherapy (median 20 vs 25 vs 120 CETC/100µl blood, p < 0.001). Surprisingly, the fraction of PD-L1 positive CETCs decreased during radiotherapy only in patients who had received neoadjuvant chemotherapy. Conclusions: Although the number of CETCs and the fraction of PD-L1 positive CETCs were higher in patients who had received neoadjuvant chemotherapy, radiotherapy reduced both measured biomarkers. In the future PD-1/PD-L1 checkpoint inhibitors after radiotherapy might have additional benefit in breast cancer patients.
Can the plasma PD-1 levels predict the presence and efficiency of tumor-infiltrating lymphocytes in metastatic melanoma patients?
Authors: Fanale Daniele , Incorvaia Lorena , Rinaldi Gaetana , Terruso Lidia , Badalamenti Giuseppe , Cucinella Alessandra , Swayden Mirna , Olive Daniel , Corsini Lidia Rita , Guarini Aurelia , Gristina Valerio , Castellana Luisa , Bono Marco , Bazan Viviana , Carreca Ignazio Ugo , Gori Stefania , Iovanna Juan , Russo Antonio
Published: May 2019

e14035

Background: The immune response to melanoma has been shown to be locally affected by presence of tumor-infiltrating lymphocytes (TILs), generally divided into brisk (infiltrating the entire base of the invasive tumor), non-brisk (infiltrating only focally) and absent. Several studies showed that greater presence of TILs, especially brisk, in primary melanoma is associated with a better prognosis and a higher survival rate. Since recent studies revealed an association between PD-1/PD-L1 expression levels and tumor response, the aim of our study was to investigate the correlation between plasma PD-1 and presence/absence/class of TILs in metastatic melanoma patients. Methods: The plasma PD-1 levels were analyzed in 28 patients with metastatic melanoma using a specific ELISA assay. The characterization of TILs in tumor tissue was performed by immunohistochemistry. Statistical analysis was assessed using t-Student and ANOVA tests. Survival curves were estimated by using the Kaplan-Meier method and log-rank test to evaluate significant differences among them. Results: 16 out of 28 patients showed the presence of TILs in primary tumor, 10 of which brisk and 6 nonbrisk. The plasma PD-1 levels were analyzed in relation to the presence/absence of TILs (p = 0.022), brisk TILs versus nonbrisk/absent TILs (p = 0.014), and brisk vs nonbrisk vs absent TILs (p = 0.032). In particular, low plasma PD-1 levels have been shown to be associated with brisk TILs in primary melanoma, intermediate values with nonbrisk TILs, and high expression with absent TILs. Although the low number of samples did not allow us to obtain a statistically significant association between the plasma PD-1 expression levels and overall survival (OS) depending on the absence or presence of TILs (brisk/nonbrisk), however the median survival of patients having brisk TILs was five months higher than other 2 groups of patients with absent and nonbrisk TILs, respectively. Conclusions: This work highlights, for the first time, the potential ability of using the plasma PD-1 levels to predict prognosis also in patients with metastatic melanoma at diagnosis for which it is not possible to identify the primary tumor.
Addition of immunotherapy in adjuvant setting could be a promising avenue of colorectal cancer (CRC) treatment, especially in patients with higher levels of programmed cell death ligand-1 (PD-L1) positive circulating epithelial tumor cells (CETC) after neoadjuvant chemotherapy.
Authors: Pizon Monika , Schott Dorothea , Pachmann Ulrich A. , Pachmann Katharina , Agrawal Siddarth , Sawicki Tomasz , Agrawal Anil Kumar , Rudnicki Jerzy , Ziolkowski Piotr , Wozniak Marta
Published: May 2019

e14036

Background: CRC is the third most frequent cancer in men and second in women worldwide. Recently, immune checkpoint molecules such as PD-1/PD-L1 have been identified as a possible target for immunotherapy in CRC and are increasingly being tested in clinical trials in combination with chemotherapy. However, the role of PD-L1 expression in CRC tumor cells and its interaction with other clinicopathologic factors remains elusive. In this study, we evaluated the impact of neoadjuvant chemotherapy on the fraction of PD-L1 positive CETCs and to compare its expression to the primary tumor. Methods: CETCs were determined from the blood of 20 patients suffering from CRC in different stages of the disease. The number of CETCs and their expression of PD-L1 were investigated using the maintrac method. In parallel, PD-L1 expression and tumor infiltrating lymphocytes (TILs) were assessed by immunohistochemistry on tissue biopsies. Results: PD-L1 expression could be assessed in all patients with detectable CETCs with a median of 50%. CETCs were more frequently found to be PD-L1 positive than tissue, and no correlation was observed between tissue and CETCs PD-L1 expression. Interestingly, patients with neoadjuvant chemotherapy had more positive PD-L1 CETCs compared to patients without chemotherapy (mean 66,7 vs. 35,2 p < 0.05), but the total number of CETCs was significantly lower in the neoadjuvant group. Also, patients with mucinous adenocarcinoma had more PD-L1 positive CETCs compared to other histological subtypes. The number of TILs was higher in patients with lymph node metastasis and larger tumors. TILs count did not differ in patients with and without neoadjuvant chemotherapy. Conclusions: Assessment of PD-L1 expression in CETCs is feasible, and CETCs are more often positive than in tissue. PD-L1 expression on CETCs was higher in patients who received neoadjuvant chemotherapy. Further studies are necessary to validate these findings.
Clinical implication of monitoring PD-L1+ circulating tumor cells (CTCs) in patients with non-small cell lung cancer (NSCLC) receiving immunotherapy.
Authors: Hoffmann Andrew Daniel , Davis Andrew A. , Iams Wade Thomas , Oh Michael S , Lentz Robert William , Chae Young Kwang , Liu Huiping
Published: May 2019

e14044

Background: CTCs not only constitute a cellular component of liquid biopsy for dynamic, longitudinal information of malignancies, but also pose a continuous threat to seed new metastasis. Detection of these cells can help identify patients with residual disease and poor prognosis. However, it is unknown if heterogeneous CTCs can be profiled to monitor cancer stemness and immunotherapy response for patients. Limited clinical data exist for monitoring CTCs for markers of cancer stem cells, such as CD44 or CD166, and immunotherapy therapeutic targets including PD-L1. We hypothesized that dynamic monitoring of CTC positivity for these markers would correlate with responses to immune checkpoint inhibitor treatment for patients with NSCLC. Methods: As part of a prospective analysis of peripheral blood cells, 30 patients with NSCLC undergoing immunotherapy (pembrolizumab or atezolizumab) or combination therapy with chemotherapy had blood samples collected at 3-4 week intervals for up to 24 weeks. Peripheral blood mononuclear cells were isolated and analyzed for CD45, to identify non-leukocyte putative CTCs, CD166 and CD44, to identify cancer stem cell markers among potential CTCs, and PD-L1. Patients were evaluated for response based on RECIST criteria. Statistical analysis was performed using one-way ANOVA on samples from each time point. Results: A decrease in PD-L1+ CD45- cells (CTCs) at the second interval was most strongly associated with initial response. In contrast, increasing number of PD-L1+ CD45- cells were found among patients with progressive disease as compared to those with stable disease or a partial response (p = 0.01). (1) CD45- cells, (2) CD44+; CD45- cells and (3) CD166+; CD45- cells demonstrated no significant association with clinical outcomes. Baseline PD-L1 level on CTCs did not correlate with the PD-L1 level on tissue. Conclusions: We report the feasibility and potential clinical significance of monitoring surface markers on CTCs in the blood before and after initiation of immunotherapy. Our findings that PD-L1+ CTCs increased among non-responders to immunotherapy warrant further validation. Testing is ongoing to evaluate how these changes predict long-term outcome to treatment for patients.
Role of the expression of PD-L1 and CD47 on circulating tumor cells (CTCs) in the prediction of outcome in metastatic breast cancer (mBC) patients.
Authors: Agelaki Sofia , Papadaki Maria A. , Tsoulfas Panormitis G , Aggouraki Despoina , Monastirioti Alexia A , Merodoulaki Katerina A , Thomopoulou Konstantina , Papadaki Chara , Mavroudis Dimitrios
Published: May 2019

e14045

Background: CTCs expressing innate (CD47) and adaptive (PD-L1) immune checkpoints may have enhanced potential to escape immunosurveillance. In the current study, we evaluated the incidence and clinical relevance of CTCs expressing CD47 and/or PD-L1 in patients with mBC. Methods: Blood was obtained from 98 mBC patients before the initiation of first-line therapy. Triple immunofluorescence staining for cytokeratins (CK), CD47 and PD-L1 was performed on peripheral blood mononuclear cells (PBMC) cytospins and 1*106 PBMCs per patient were analyzed using the Ariol microscopy system. CD47 and PD-L1 expression levels on CK+ CTCs were normalized using MDA.MB.231 breast cancer cells as controls. Results: CK+ CTCs were detected in 22 out of 98 patients (22.4%) (total CTC No: 43; range: 1-12). High CD47 and PD-L1 expression was identified in 41.9% and 11.6% of CTCs, respectively, whereas in 9.1% of CTCs, high expression was observed for both markers. CTCs with high expression of at least one marker ( CD47high and/or PD-L1high) were identified in 11.2% of patients, albeit with a differential distribution among patients with triple-negative, hormone receptor-positive and HER2-positive primary tumors (27.3%, 12.5% and 0%, respectively, p = 0.040; Chi-square test). CD47high and/or PD-L1high CTCs were associated with disease progression (27.8% vs 5.6%, p = 0.005; Chi-square test) and shorter PFS [median: 5.8 (4.1-7.5) vs 13.3 (11.4-15.2) months, p = 0.010; Kaplan-Meier], whereas the detection of PD-L1high CTCs only was correlated with reduced OS [median: 23.8 (5.8-41.8) vs 35.7 (29.9-41.4) months, p = 0.043]. Multivariate Cox-regression analysis revealed that CD47high and /or PD-L1high CTCs predicted for increased risk of relapse [HR: 2.7 (95% CI: 1.3-5.7); p = 0.009], whereas PD-L1high CTCs were associated with high risk of death [HR: 4.8 (95% CI: 1.4-16.6); p = 0.011]. Conclusions: The immune checkpoints CD47 and PD-L1 are co-expressed in a subset of CTCs in mBC patients. The detection of high CD47 and/or PD-L1 expression on CTCs is associated with triple-negative tumors, disease progression and poor patient outcome and could serve for the refinement of prognosis in mBC. Patients bearing this CTC population may benefit from anti-CD47 and anti-PD-L1 immunotherapy strategies.
Correlation of HERVK with efficacy on anti-PD-1/PD-L1 therapy in a pilot metagenomics study on Chinese advanced lung cancer patients.
Authors: Tao Haitao , Hu Yi
Published: May 2019

e14046

Background: Patient selection for PD-1 inhibitors in NSCLC is still based on imperfect screening biomarkers, including PD-L1 expression and tumor mutational burden(TMB).There are several reports show certain bacteria or virus infections are correlated with efficacy and prognosis on PD-1 inhibitors treatment. In order to explore more potential biomarkers for anti-PD-1 therapy, we investigate whether we can discover microorganisms in Chinese lung cancer patients and whether certain microorganism are correlated with efficacy or prognosis on anti-PD-1 therapy using metagenomics technology. Methods: In this study, 30 advanced lung cancer patient were enrolled, including 15 cases of adenocarcinoma, 12 cases of squamous carcinoma, 2 cases of Small cell lung cancer. 10 patients received PD-1/PD-L1 inhibitors as first-line therapy and other patients as second-line therapy. Among these 30 patients, 15 PR,1 CR, 10 PD, 4 SD according to RECIST 1.1. Plasma cell-free DNA (cfDNA) were collected from these patients prior to anti-PD-1 treatment and whole-genome sequencing of cfDNA at average coverage 5X was performed on Illumina NovaSeq in 2 ×150bp length model. A bioinformatics pipeline composed of 4 tools: BWA/Sambama/Kraken2/Bracken, was constructed to analyze the sequencing data. Non-human derived reads were extracted by sambama from the bam file that generated by BWA. Bacteria/Virus abundance in each sample was estimated through Bracken, followed by bac/vir database alignment in Kraken2. Results: In our pilot study, 4 patients were selected for metagenomics sequencing(1CR, 1PR, 2PD). Interestingly, over 100 different microbial sequence were detected in every patient. Compare microorganism in CR+PR patients to PD patients, Human endogenous retrovirus K (HERVK) were quite abundant in CR/PR patients but not in PD patients: 5.6 times more abundant in CR/PR patients comparing to PD patients. (HERVK abundance: CR:PR:PD = 1.20% : 0.32% : (0.17%, 0.10%)). HERVK is a family of human endogenous retroviruses associated with certain malignant tumors, studies have indicated that HERVK may trigger an antigen specific immune response for certain cancer patients. Conclusions: Metagenomics sequencing on plasma cfDNA could be a potential biomarker discovery approach to identify prognostic biomarkers for anti-PD-1 therapy. HERVK are correlated with efficacy on anti-PD-1 treatment in our pilot study, however more thorough investigation need to be conducted to confirm whether HERVK can be served as efficacy biomarkers for anti-PD-1.
Association of relative neutrophilia with a distinct immunoinhibitory milieu in non-small cell lung cancer.
Authors: Mitchell Kyle Gregory , Diao Lixia , Tran Hai T. , Negrao Marcelo Vailati , Karpinets Tatiana , Wang Jing , Parra Cuentas Edwin Roger , Corsini Erin M , Reuben Alexandre , Federico Lorenzo , Bernatchez Chantale , Vaporciyan Ara A. , Swisher Stephen , Cascone Tina , Wistuba Ignacio Ivan , Heymach John , Zhang Jianjun , Gibbons Don Lynn , Haymaker Cara L. , Sepesi Boris
Published: May 2019

e14047

Background: Elevated neutrophil-to-lymphocyte ratio (NLR) has been associated with poor prognosis in non-small cell lung cancer (NSCLC); the biological underpinnings of this observation have not been fully elucidated. We examined the relationships between peripheral neutrophil counts (PMN), NLR, circulating cytokines and angiogenic factors (CAF), and tumor microenvironment (TME) features in NSCLC. Methods: 150 patients with resectable NSCLC were enrolled in an immunoprofiling project. A panel of 43 CAFs was used to analyze preoperative plasma samples. Chemotherapy-naïve patients with CAF and a complete blood count ≤30 days preoperatively were included (n = 66; Table). For a subset, transcriptional signatures (MCP-counter, n = 50) and flow cytometry (n = 19) were used to identify TME phenotypes. Results: Increased PMNs were associated with increased pro-inflammatory CAF such as IL-1b (r = 0.392) and IL-6 (r = 0.339), as well as Th17/Tc17 associated CAF IL-17A (r = 0.320) and TNF-a (r = 0.368). Elevated NLR was inversely correlated with the lymphocyte activation marker soluble CD27 (r = -0.320, p = 0.009). This negative association was mirrored in the TME, as tumor neutrophil signatures were inversely correlated with a local IFN-g gene signature (r = -0.626, p < 0.001). Interestingly, a Th17/Tc17 peripheral signature (elevated IL-17A) was associated with an enrichment of CD8+TIM3+ cells (r = 0.623, p = 0.042) in the tumor. While this requires confirmation in a larger cohort, this correlation provides a potential rationale for targeting TIM3 in this population. Upon analysis of clinical characteristics, peripheral PMNs and NLR were higher among patients with squamous histology (PMN p = 0.009; NLR p = 0.034) and positively correlated with tumor size (PMN r = 0.344, p = 0.004; NLR r = 0.363, p = 0.003). Conclusions: A relative neutrophilia in NSCLC patients is associated with an inflammatory milieu suggestive of a Th17/Tc17 presence and decreased lymphocyte activation that is reflected within the TME. Further investigation is needed to define the role of NLR as a predictive biomarker and to identify whether neutrophils or Th17/Tc17 T cells could serve as a therapeutic target to improve immunotherapy response in NSCLC.[Table: see text]
A relationship-based approach to improving the clinical trial experience.
Authors: Helou Sarah MacKay , Gadberry Camber W. , Zahara Nicholas , Nickols Claire , Vidhyarkorn Tanya , Otap Daniel , Russell Janice , Piggee Jessica , Scher Catherine , Estrabaut Myriam , Churchill Sarah , Tran San
Published: May 2019

e14055

Background: The rapidly evolving field of Cancer Immunotherapy (CIT) has mobilized biopharma to develop promising new treatments while at the same time created a need to expedite the pathway of those new therapies from bench to bedside. This race to bring innovative treatments to patients has engendered heightened levels of competition for the attention of and partnership with the best research institutions in the biopharma industry. Methods: Recognizing this reality, Roche created the immunotherapy Centers of Research Excellence (imCORE) Network: a network of leading cancer immunotherapy centers worldwide. The vision of this network was to partner with sites to streamline the operational delivery of the Roche Genentech portfolio while deepening the collective understanding of the science to prioritize and accelerate the most promising new treatment options for our patients. Creation of a network was not a novel idea, but Roche set out to build a network unique to the biopharma industry – one which was relationship-based where the site/sponsor relationship serves as the foundation for driving both collaborative scientific investigations and operational efficiencies in support of Roche’s clinical trials. While the identification and optimization of promising candidate drugs in the preclinical and translational settings are critical first steps in the drug development process, the initiation and operationalization of early phase clinical trials are equally important in the multidimensional cycle of drug development. Results: See Conclusions. Conclusions: This white paper will provide an overview of imCORE’s origins with a focus on the strategic importance of the site-sponsor relationship and the mutual value that is derived from the combined benefits of these relationships and in conducting early clinical trials more efficiently across Roche’s CIT portfolio. Further examination of communication and operational improvement to support the network will be highlighted as key drivers in creating and sustaining meaningful site/sponsor relationships that better facilitate the operationalization of clinical trials and ultimately brings novel therapies to patients in a more efficient manner than in the past.
Effect of mandatory research biopsy requirements on initiation of clinical trials.
Authors: Cheng Jonathan Hin , Tiulim Justin Wayne Wong , El-Khoueiry Anthony B. , Nieva Jorge J.
Published: May 2019

e14057

Background: There has been an increasing requirement for fresh tumor tissue to enroll in clinical trials in order to look for specific biomarkers. This has been shown to increase screening duration and increase screen failure rates. It was important to corroborate these results in other centers. Methods: This study is a non-randomized retrospective analysis of patients in one subset of patients seen by research nurses who operated in the standard head/neck and lung team not including patients in the phase 1 program. All patients were enrolled in clinical trials from January 16, 2013 to May 28, 2018 at USC Norris Comprehensive Cancer Institute in Los Angeles. Patients who were required to give fresh research biopsies prior to intervention were part of the research biopsy group. Results: In total, 76 patients were analyzed in this study. 33 patients were in the research biopsy group and 43 patients were in the no biopsy group. Trials that required a fresh biopsy had a longer median screening duration (30 vs 14 days) than trials that did not require a biopsy (p < .0001). Conclusions: Our study shows that requiring biopsies prior to clinical trial treatment results in a statistically significant delay in treatment. The informed consent forms that were part of clinical trials involving mandatory research biopsies did not reflect this delay in treatment. However, these delays did not result in a statistically significant decrease in number of days on trial or days until progression of disease.
Tislelizumab, an investigational anti-PD-1 antibody, combined with chemotherapy as first-line treatment for lung cancer in Chinese patients.
Authors: Wang Jie , Zhao Jun , Wang Zhijie , Ma Zhiyong , Cui Jiuwei , Shu Yongqian , Liu Zhe , Cheng Ying , Leaw Shiangjiin , Xia Fan , Wu Yanjie , Liang Liang
Published: May 2019

e14067

Background: Tislelizumab, an investigational anti-PD-1 antibody, was engineered to minimize binding to FcγR on macrophages in order to abrogate antibody-dependent phagocytosis, a mechanism of T-cell clearance and potential resistance to anti-PD-1 therapy. Previous reports showed tislelizumab was generally well tolerated and had antitumor activity in patients (pts) with advanced solid tumors; 200 mg IV Q3W was established as the RP2D. Methods: This phase 2 clinical trial (NCT03432598) assessed tislelizumab (200 mg Q3W) with platinum (plt)-based chemotherapy (Q3W) as first-line treatment for Chinese pts with advanced lung cancer. All pts received tislelizumab + plt doublet (4–6 cycles) until disease progression. Nonsquamous (nsq) NSCLC pts received pemetrexed (PMX) + plt (4 cycles) followed by PMX maintenance; squamous (sq) NSCLC pts received A) paclitaxel (PXL) + plt or B) gemcitabine + plt; SCLC pts received etoposide + plt. Tumor response (RECIST v1.1) and safety/tolerability were evaluated. PD-L1 expression was retrospectively assessed with the VENTANA PD-L1 (SP263) assay. Results: As of 15 Oct 2018, 54 pts (median age 61 yr; 74% male; 72% current/former smokers; 31% with ≥10% PD-L1 expression on tumor cells) received tislelizumab; 24 pts remain on treatment. Confirmed PR was observed in 36 pts and most occurred within the first 2 assessments. Other efficacy estimates (eg, PFS) are maturing. Grade ≥3 AEs occurring in > 15% of pts were decreased neutrophil counts (n = 25) and anemia (n = 9); immune-related AEs occurring in ≥2 pts were decreased triiodothyronine, hyperthyroidism, hypothyroidism, and pyrexia (n = 2 each). One sq-NSCLC pt ( A) experienced fatal myocarditis/myositis after 1 cycle; other AEs resolved with tislelizumab interruption (n = 30), discontinuation (n = 4), or other appropriate treatment. Conclusions: Tislelizumab in combination with standard of care plt-based chemotherapy was generally well tolerated and demonstrated antitumor activity. Clinical trial information: NCT03432598. [Table: see text]
Burden of copy number alterations predicts outcomes in immune checkpoint blockade treated gastrointestinal cancer patients.
Authors: Lu Zhihao , Chen Huan , Li Shuang , Jiao Xi , Wu Lihong , Zhang Henghui , Shen Lin
Published: May 2019

e14068

Background: Despite the great achievements made in immune checkpoint blockade (ICB) in cancer therapy, how to identify patients who may benefit from ICB still remains one of the central questions, especially in gastrointestinal (GI) cancer. Methods: To address this, we analyzed FFPE tumor specimens from 73 patients with metastatic GI cancers who were treated with ICB. All patients were randomly assigned into discovery (60%) and validation (40%) cohorts. Overall, tumor mutation burden (TMB) and copy number alterations (CNAs) were determined by using the whole-exome sequencing platform. FFPE samples of 65 patients were analyzed via a multiplex RNA immune oncology sequencing panel. Results: Here we show that lower burden of copy number alteration (CNA) was observed in responders to immunotherapy in both discovery and validation cohorts. More importantly, lower burden of CNA in GI cancers were associated with improved objective response, clinical benefit and overall survival. Efficacy also correlated with the higher TMB. Of note, a combinatorial biomarker of TMB and burden of CNA may better stratify responders from patients received immunotherapy. In addition, patients with lower burden of CNA revealed increased IFNγ and expanded immune signatures in our GI patient cohort and TCGA cohorts as well. Conclusions: Our results suggest that burden of CNA may expand the predictive and prognostic value of genomic determinants in identifying potential responders with GI cancer to immune checkpoint blockade therapy.
Review of therapeutic approaches for B-cell malignancies with immune checkpoint blockade and chimeric antigen receptor t-cell therapies: Development, benefits, and limitations.
Authors: Hays Priya
Published: May 2019

e14069

Background: Immune checkpoint inhibitors and Chimeric Antigen Receptor (CAR) T-cell therapies have emerged as approaches to treat B-cell malignancies. Methods: PubMed/NCBI/MEDLINE databases were accessed with keywords "immune checkpoint inhibitors and B-cell malignancies" "CAR T-cell and B-cell malignancies", and various permutations including "clinical data" "toxicities", "reviews", "quality of life", and "adverse effects". Results: The first-in-class approved immune checkpoint inhibitor was ipilimumab, which is a fully humanized mAb that blocks the immunosuppressive signal by cytotoxic T-lymphocyte antigen. Thereafter, nivolumab was also approved for use in the treatment of Hodgkin's lymphoma in 2016. In phase I, open-label, dose-escalation, cohort-expansion study, patients with relapsed or refractory B-cell lymphoma received the anti-PD-1 monoclonal antibody nivolumab. Eighty-one patients were treated and drug-related adverse events occurred in 51 (63%) patients. Objective response rates were 40%, 36%, 15%, and 40% among patients with follicular lymphoma and other hematologic malignancies. Clinical trial results describing CD19-targeted CAR T-cell therapy of patients with relapsed B-ALL were published in 2015. In this study, all five patients treated with CAR T cells achieved minimal residual disease negative complete remission. Updated results describing the treatment of 16 patients with relapsed or refractory B-ALL treated with CAR T cells were published: the overall CR rate in this trial was 88% and 12 of 14 patients were classified as minimal residual disease negative. 44% of these patients went on to standard-of- care allogeneic hematopoietic stem cell transplant. Initial studies also reported potent anti-leukemic effects of CD19 CAR T cell therapy in three patients with refractory chronic lymphocytic leukemia where two of the three patients achieved MRD-CR. Infused CAR T cells proliferated up to 10,000-fold and persisted in recipients for at least 6 months and shown to retain antitumor activity after six months. Costs for CAR T-cell therapies remain exorbitant, reaching over $1M (USD) per patient. Conclusions: Clinical data reveal safety and efficacy, and also associated toxicities for both approaches.