bluuebunny/crossref_metadata_2025 huggingface dataset:
Outcomes and toxicities in sarcopenic patients with metastatic solid tumors treated with check point inhibitors.
Authors: Haik Laura , Gonthier Aurore , Frison Eric , Gross-Goupil Marine , Domblides Charlotte , Veillon Remi , Quivy Amandine , Ravaud Alain , Daste Amaury
Published: May 2019

e14166

Background: Evidence suggests that sarcopenia is a significant predictor of worst outcomes and treatment toxicities in metastatic solid tumors patients (pts). We investigated whether sarcopenia was associated with outcomes and severe toxicities (ST) in patients treated with check point inhibitors (CPI). Methods: All consecutive metastatic solid tumors patients treated with CPI from January 2013 to December 2017 in Bordeaux University Hospital (France) were retrospectively reviewed. Sarcopenia was evaluated using computed tomography (CT) obtained within 1 month before treatment initiation. Skeletal muscle mass index (SMI) were measured from axial L3 CT section. Sarcopenia was defined as SMI ≤ 52.4 cm2/m2 for males and SMI ≤ 38.5 cm2/m2 for women. Clinical characteristics included Performance Status (PS) and anthropometric parameters. ST included grade III-IV according to NCI-CTC v4.0 and any grade of toxicity leading to treatment interruption. Results: Among 261 pts included, 179 (69%) had metastatic lung cancer and 151 (58%) had previously received one line of treatment. Median age was 61 years old and there were 198 males (76%). PS status was 1 in 55% of pts. CPI treatment was: anti PD1, anti PD-L1, anti CTLA-4 and CPI combination in 192, 43, 3 and 20 pts, respectively. The prevalence of sarcopenia was 64%. At inclusion, sarcopenia was associated with poor PS (p = 0.002), anorexia (p < 0.001), lower albumin (p = 0.002) and hemoglobin (p < 0.001). Objective Response Rate (ORR) was 28% in sarcopenic pts and 26% in non-sarcopenic pts. Median progression free survival (PFS) was 6.9 and 7.6 months in sarcopenic pts and non-sarcopenic pts respectively (p = 0.5). In multivariate analysis, sarcopenia was not associated with PFS or ORR. Median overall survival (mOS) was 10.2 months in lung cancer, without significant difference between sarcopenic and non-sarcopenic patients (mOS: 9 months and 17 months respectively, p = 0.06). In general population, ST occurred in 36 (14%) pts; among which 23 (64%) presented sarcopenia. In multivariate analysis, sarcopenia was not associated with ST (OR = 1.16, p = 0.71). Conclusions: Sarcopenia was not associated with outcomes or toxicities in metastatic solid tumors patients treated with CPI in this study.
Immunotherapy efficacy and patients' age: A systematic review and meta-analysis.
Authors: Duan Jianchun , Cui Longgang , Zhao Zhengyi , Wang Guoqiang , Chen Shiqing , Bai Yuezong , Cai Shangli , Xiong Lei , Zhao Xiaochen , Wang Zhijie , Wang Jie
Published: May 2019

e14168

Background: Immunotherapy has revolutionized the treatment of cancer, however, little is known about the effect of patients’ age on the efficacy of immune checkpoint inhibitors. We did a systematic review and meta-analysis to assess the heterogeneity of immune checkpoint inhibitor efficacy between young and elder patients in pan cancer. Methods: We systematically searched PubMed, Cochrane library, and Embase from January 2000 to November 2018 for randomized controlled trials that had available hazard ratios (HRs) for death according to patients’ age. We also reviewed abstracts and presentations from all major conference proceedings. Retrospective studies and trials that compared anti-PD-1/PD-L1 with other immunotherapies were excluded. The primary outcome was the difference in overall survival (OS). We calculated the pooled overall survival HR and 95% CI in young and elder patients using a random-effects model. Results: A total of 3365 publications were retrieved through initial literature search, 19 randomized controlled trials involving 12276 patients with solid tumors were included for this meta-analysis. PD-1 inhibitors exhibited significantly improved OS over PD-L1 inhibitors in younger patient population (HR 0.67, 95% CI 0.53-0.85), while no differences have been observed in elder population (HR 0.89, 95% CI 0.70-1.13). Subgroup analysis showed that PD-L1 inhibitors plus chemotherapies led to significantly improved overall survival (OS) in elder population (> = 65 years old) over that in younger population (< 65 years old) (HR 1.29, 95% CI 1.01-1.64). However, the opposite tendency was observed with PD-1 inhibitors plus chemotherapies, showing that the younger population obtained better OS benefit compared to the elder population (HR 0.69, 95% CI 0.47-1.01). Conclusions: In general, PD-1 inhibitors exhibited better clinical performance for survival outcome over PD-L1 inhibitors in younger patients. When combined with chemotherapies, PD-1 inhibitors and PD-L1 inhibitors showed distinct efficacy tendency across different age groups, indicating that age should be taken into account when making decisions about treatment strategies. Future studies and explorations of the underlying mechanisms are needed for the further optimization of treatment strategies in clinical practice.
Defining real-world criteria for immune-related adverse events (irAEs).
Authors: SINGH PRABHSIMRANJOT , Brito Amanda , Abdulnour Raja-Elie E , Grover Shilpa , Yenulevich Eric , Stuver Sherri Oliver , Kehl Kenneth L. , LeBoeuf Nicole R. , Jacobson Joseph O. , Rahma Osama E.
Published: May 2019

e14172

Background: Management of Immune checkpoint inhibitors (ICIs) associated irAEs requires accurate diagnosis and severity measurement. However, outside of clinical trials, there is no clear definition of irAEs to guide treatment. Methods: We established a dedicated immune toxicity inpatient service (ITox) for patients admitted with irAEs using institution-developed guidelines adapted from NCCN and ASCO. We developed definitions for ICI associated hepatitis, colitis and pneumonitis and created algorithms for defining an irAEs as definite, likely, possible, or unlikely. Algorithm fields were completed twice, by two independent reviewers including medical oncology specialists and toxicity specialists (a pulmonologist or gastroenterologist) to evaluate the accuracy of the algorithm in defining irAEs. Results: From June to November 2018, 65 patients were admitted to iTox with suspected irAEs. Various cancers patients treated with ICI were included; with lung and head and neck cancers the most common (36.9%). Pembrolizumab was the most common ICI (47.7%). ICI in combination with non-checkpoint inhibitor treatment was used in 23 (37.7%) and concurrent radiation therapy in 5 patients (7.7%). Forty (71.4%) patients discontinued ICI due to toxicity. Fifty-one patients were evaluable with irAEs: colitis (19); pneumonitis (24) and hepatitis (8). Multiple irAEs were reported in 17 patients. When applied to clinical cases, our algorithms generated a toxicity definition in 63% of cases, with which reviewers agreed most of the time (84-100%) (Kappa 0.7-1.0). Concordance between the two reviewers was 75-87% (Kappa 0.55-0.82) (Table). Conclusions: Our algorithms generated irAEs definitions with fair inter-rater reliability. These findings have real-world implications; accurate diagnosis of irAEs is essential for proper management. Defining irAEs outside the clinical trial context remains an important challenge. [Table: see text]
Toxicities of single agent and combination immune checkpoint inhibitors in patients with autoimmune diseases.
Authors: Cytryn Samuel , Efuni Elizaveta , Sandigursky Sabina
Published: May 2019

e14176

Background: Autoimmunity is associated with increased risk of malignancy. However, patients with pre-existing autoimmune diseases (AIDs) have been excluded from trials of immune checkpoint inhibitors (ICIs) known to cause immune activation and lead to immune related adverse events (irAEs). Data is limited on the safety and efficacy of these agents when used in AID patients and physicians are therefore wary to use them in this at-risk population. Methods: We conducted a single institution retrospective study to evaluate the safety and efficacy of ICI therapy in patients with pre-existing AIDs from 2011 to 2018. Primary endpoints were irAEs and AID flares. The secondary endpoint was overall survival (OS). Results: Of 84 total patients, 70 (83%) received ICI monotherapy and 14 (17%) received combination therapy. AIDs included: rheumatic 40 (48%), dermatologic 25 (30%), endocrine 16 (19%) and gastrointestinal 14 (17%) diseases of which 18 (21%) were on immune suppression. Combination therapy was associated with higher rates of severe grade 3-4 irAEs in 5/14 (36%) patients versus 8/70 (11%) with monotherapy (p = 0.022). ICIs were discontinued due to irAEs in 10/84 (12%) of all patients; 4/14 (29%) in the combination group and 6/70 (9%) in the monotherapy group (p = 0.057). While 32 patients (38%) had at least one AID flare, ICI was only permanently discontinued in 2 patients. Flare rates were higher for combination use: 8/14 (57%) compared to 23/70 (33%) for monotherapy (p = 0.086). Combination therapy was associated with higher median OS 27.8 months [95% CI 11.4-44.3] compared to monotherapy 12.3 months [95% CI 9.7-14.8] (p = 0.0007). OS had a positive correlation with flare (p = 0.007) and severe irAEs (p = 0.037). Conclusions: Our data suggest that rates of irAEs in patients with pre-existing AIDs are similar to those reported in the literature with single agent ICIs. While risk of severe irAEs and flares is increased with combination therapy, they are associated with overall survival. In our cohort, irAEs and flares were manageable and in most patients did not require permanent discontinuation for monotherapy or combination therapy suggesting that ICIs should be offered to this population, albeit with caution.
An effective and accurate algorithm for measuring the tumor mutation burden of cancer patients plasma.
Authors: Liu Bo , Zhou Shuang , Cai Yu Hang , Yang Ying , Ye Mingzhi , Shi Tai Ping
Published: May 2019

e14177

Background: Tumor Mutation Burden (TMB) is deemed to be one of the effective genomic biomarkers that are used to predict the response to PD-1/PD-L1 blockade immunotherapy. As an exciting biomarker for diagnose and an alternative assess method for tissue TMB (tTMB), blood TMB (bTMB) is measured to further distinguish the patients who may benefit from immunotherapy. However, there is no recognized rule to calculate bTMB so far. Methods: In this study, we developed an in-house algorithm called PTAB to calculate bTMB of plasma, all credible single nucleotide variants and insertion-deletions whose allele frequency is greater than 1% are selected in PTAB without synonymous mutations and drive mutations. We also constructed a 636 genes panel which region size is 1.95Mb to identify all the associated mutations, due to the high costs and long turnaround time of WES. Results: Meanwhile, we assessed a cohort of 50 lung cancer plasma samples and paired tissues to ensure the consistency of different algorithm used for tissue and plasma. The results show that Pearson correlation coefficient (R2) between bTMB and tTMB in 50 patients is 0.7128. In addition, we found that the consistency of lung squamous carcinomas obviously higher than that of lung adenocarcinoma (0.8932 vs. 0.5947). We also described the distribution of bTMB and tTMB value of Chinese patients, including 594 tissue samples and 561 plasma samples from 8 types of cancers, including lung cancer, colorectal cancer, breast cancer, et al. The overall comparative analysis of 1155 previous data suggested that the distribution modes of bTMB and tTMB were almost same in certain cancer type, though the level of bTMB was generally lower than that of tTMB. Conclusions: Preliminary data highlight that PTAB is an effective and accurate algorithm for measure the TMB of plasma. The applicability of bTMB testing on liquid biopsy needs further investigation and may be clarified within the ongoing clinical trials.
Anti-HER2/Neu antibody therapy can reduce the immunosuppressive activity of MDSCs in breast tumor model.
Authors: Kim Il-Hwan
Published: May 2019

e14181

Background: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous cell populations that play a critical role in tumor associated immune suppression. MDSCs consist of immature cells with myeloid lineage markers CD11b and Gr1 in mice and accumulate in large numbers in tumor bearing mice including HER2/neu+ breast cancers. This study was conducted to test whether anti-neu antibody treatment can affect on MDSC populations and immunosuppressive functions. Methods: Female BALB/c mice(5–6weeks of age) were used. The TUBO mammary carcinoma cell line was cloned from a spontaneous mammary tumor in a BALB Neu Tg mouse, as cell lines and reagents. Tumor-infiltrating lymphocyte (TIL) populations were evaluated by flow cytometry. The RT2 Profiler PCR array was used for cytokine expression analysis according to the manufacturer’s instructions. Results: Total number of MDSC in tumor and spleen were decreased within 3 days after anti-neu antibody treatment, but percentage of MDSC in tumor (day2) was reduced more quickly than percentage of MDSC in spleen (day3). The relative populations of MDSC in CD45.2+ cells were significantly decreased such as MO-MDSC. Immunosuppressive activity of MDSC was also reduced by anti-neu antibody treatment. MDSC reduced by anti-neu antibody was due to inhibition of the immunosuppressive factors Arg1 and IDO. In addition, combination therapy with 5-FU may enhance MDSC reduction. Conclusions: MDSC can be a potential therapeutic target in anti-neu antibody treated therapeutic model and the effect of anti-neu antibody could be further improved using other therapeutic treatment to target MDSC.
The influences of preoperative metformin on immunological factors in early breast cancer.
Authors: Takahiro Takahiro , Shien Tadahiko , Taira Naruto , Kochi Mariko , Iwamoto Takayuki , Doihara Hiroyoshi , Udono Heiichiro , Toyooka Shinichi
Published: May 2019

e14182

Background: Metformin is one of the most commonly prescribed drugs for type 2 diabetes, and some reports have suggested that metformin may reduce cancer risk. Diabetics treated with metformin have a 23% reduction in the risk of cancer, including breast cancer. In addition, it is reported that the breast cancer patients with metformin treatment for diabetes showed favorable prognosis compared with those without metformin treatment. However, the mechanism underlying the positive effects of metformin on cancer treatment remains unclear. Methods: We conducted a prospective study to evaluate the effect of preoperative metformin on early breast cancer patients. The patients took a daily dose of metformin orally for 7 to 21 days before surgery. We evaluated the effects on immunological factors (TILs, CD4+, CD8+, PD-L1 and ALDH1) by comparing core needle biopsies (CNB) obtained before surgery with surgical specimens. Results: Seventeen breast cancer patients were enrolled in this prospective study and administered metformin before surgery, during the period from January to December 2016. We analyzed 59 patients who received surgery during the same period as a control group. In the control group, there was no significant difference in TILs between CNB and surgical specimens (Rs = 0.63). In the metformin group, TILs were negative in CNB and surgical specimens of 15 (88%) and 8 (48%) cases, low in 2 (12%) and 8 (48%), and intermediate in 0 and 1 (6%), respectively. These TILs increases were confirmed in 5 (29%) patients (p = 0.09), while a decrease was confirmed in 2 (12%). The expressions of CD4+ and CD8+ by TILs were increased in 41% and 18% of surgical specimens, respectively. (p = 0.02, p = 0.09) However, there was no statistically significant difference in these immunological factors and PD-L1 or ALDH1 expression between before and after metformin. Conclusions: In our small cohort, preoperative metformin administration shows positive impact on CD4 positive lymphocytes significantly and has tendency of increasing both TILs and CD8 positive lymphocytes. However, we should keep in mind that our sample size is small. Further study is necessary to uncover the mechanisms of favorable effects of metformin on breast cancer patients.
Use of PD-L1 and ICOSL to discriminate innate and adaptive dendritic cells in the head and neck cancer microenvironment.
Authors: Hoffmann Caroline , Surun Aurore , Noel Floriane , Michea Paula , Grandclaudon Maximilien , Faucheux Lilith , Sirven Philemon , Klijanienko Jerzy , Badois Nathalie , Lesnik Maria , Choussy Olivier , Kamal Maud , Lecerf Charlotte , Le Tourneau Christophe , Guillot-Delost Maude , Soumelis Vassili
Published: May 2019

e14184

Background: Inflamed head and neck squamous cell carcinomas (HNSCC), have a better prognosis independently of the treatment modalities, and have an increased response rate to PD(L)1 blockade. Dendritic cells (DC) infiltration is associated to T cell infiltration and DC are critical in the immune response to cancer, but little is known on HNSCC infiltrating DC. The objectives were to determine the differences between DC infiltrating inflamed vs non inflamed tumors, to understand which mechanisms modulate DC phenotypes and which are the associated functions. Methods: 22 untreated HNSCC were characterized by flow cytometry for their immune infiltrate. RNA sequencing of sorted tumor infiltrating myeloid subsets and blood DC were analyzed for differentially expressed genes. The TMod database (Grandclaudon et al.), testing the effect of various stimuli on DC and subsequently on co-cultured T cells, was used to decipher DC and T cell modulation. Results: The level of CD3 infiltration was positively associated to the level of DC infiltration and PDL1 expression on myeloid cells, and negatively to the level neutrophils and of ICOSL expression on myeloid cells. PDL1high ICOSLneg DC from inflamed tumors were matched with the “innate DC” phenotype described in vitro, highly efficient at secreting cytokines and chemokines but poorly activating naïve CD4 T cells. DC from non-inflamed tumor resembled immature medium DC. A third type of DC, “adaptive DC”, expressing high levels of ICOSL and low of PDL1, poorly efficient at cytokine and chemokine secretion, but highly efficient at activating T cells is was described in vitro, but not found in tumors. RNAseq analysis confirmed this innate polarization and highlighted that those cells present a mature and NFkB activated phenotype. Conclusions: Inflamed HNSCC contain stimuli able to induce the recruitment and the differentiation of DC into mature, NFkB activated “innate” DC, highly efficient at secreting cytokines and chemokines but poorly activating naïve CD4 T cells. These data partly explain the superiority of CPS over TPS for the prediction of response to PD(L)1 blockade. More importantly, the stimuli identified here may be used as adjuvants to induce this DC « innate » maturation in non-inflamed tumors, but should only be used in combination with anti-PD(L)1 immunotherapy.
Demographic stratification of inflammatory signature in lung cancer patients in North Carolina: A prospective cohort study.
Authors: Sharma Nitika , Walker Paul R. , Stroud Chipman Robert Geoffrey , Cherry Cynthia R.
Published: May 2019

e14200

Background: Lung Cancer remains the major cause of cancer related mortality in the state of North Carolina. There is a growing body of evidence that implicates inflammation as a mechanism of disease progression and reduced survival in patients with advanced cancer (Laird et al, Oncologist 2013). Smoldering inflammation in the tumor microenvironment regulates and escalates cancer invasion, angiogenesis and immune surveillance escape (Balkwill and Mantovani, Lancet 2001). We investigated the predictive value of inflammatory signature according to social stratification of cancer patients using Modified Glasgow Prognostic Score (mGPS). mGPS is a composite inflammatory score based on CRP and serum albumin with proven prognostic and predictive value in various tumor types. Methods: A prospective observational single institutional study was conducted whereby serum albumin and CRP were drawn at baseline for 333 patients with diagnosis of cancer regardless of stage from 30 counties in Eastern North Carolina. The mGPS score was compared according to rural urban divide and occupational regional exposure of various counties stratified per US Census Data. Results: Lung cancer was the predominant cancer type in 93% of patients. The mGPS of zero in Urban vs Rural counties was noted in 36% and 24% patients respectively. The mGPS score of two in Urban vs Rural counties was noted in 26% and 41% respectively. The mGPS of two in areas of hog farming, cattle farming and wet waste lands was seen in 41%, 38% and 43% respectively (p = 0.0019). The mGPS of zero was seen in 24%, 20% and 27% respectively (p = 0.0008). Conclusions: This study suggests a strikingly unfavorable inflammatory signature in rural population as well as areas of hog farm, cattle farm and wet waste lands. The hog and poultry operations heighten the harmful effect on waterways and can adversely affect the inflammatory signature, hence the tumor biology. This underscores additional interventions in these high risk populations that can have significant implications for quality of life and survival, especially in the era of immunotherapy.
The interplay of adiposity, metabolic factors, and tumor infiltrating lymphocytes in an East Asian breast cancer cohort.
Authors: Chen I Chun , Kuo Chun-Wei , Hsueh Tsu-Hsin , Lu Yen-Shen , Lin Ching-Hung
Published: May 2019

e14207

Background: Breast cancer is rapidly emerging in East Asian countries in recent 2 decades. Unlike western countries in which obesity are quite prevalent and contributes to BC patients, the body mass index (BMI) of BC patients in East Asian countries are usually normal to no more than overweight. In our previous study, we noticed a 30% incidence of crown-like structure of breast (CLS-B) in our BC patients. Despite a lower BMI in this East Asian cohort than the US cohort, the body fat percentage (BF%) is significantly higher in the East Asian The roles of tumor infiltrating lymphocytes (TILs) in BC are inconslusive in different disease contexts. This study aims to decipher the interplay of adiposity, metabolic factors, and TILs in a BC cohort from an East Asian country. Methods: Breast cancer patients from National Taiwan University Hospital were prospectively enrolled into the protocol. Clinicopathologic factors were collected from electronic medical records. Fasting serum collected prior to surgery was used to quantify metabolic factors, including lipid and glucose profiles. Adiposity, including BF% and BMI were quantified by bioimpedance analysis method prior to surgery. Immunohistochemical staining from formalin-fixed, paraffin-embedded BC tumor samples were done to profile TILs by pathologist. CLS-B was stained and interpreted by pathologists. The correlation of TILs with different clinicopathologic factors, adiposity, or relapse-free survival (RFS) were analyzed. Results: 72 BC patients were included in this analysis. The density of CLS-B positively correlates with tumor infiltrating lymphocyte (TIL) percentage ( p = .042), higher BF% correlates with higher regulatory T cells ( p = .026) and tumor associated macrophages (TAM, p = .010), high glucose level is associated with higher CD56 ( p = .040) and higher TAMs ( p < 0.0001). Higher BMI is also associated with higher TAMs ( p= .005). In multivariate analysis, TAMx M2 macrophage is an independent predictor for RFS. Higher TAM level is associated with shorter RFS. Conclusions: Higher TAMs are associated with metabolic alterations such as higher adiposity, higher glucose level, and higher BMI. This result suggests a potential interplay between higher TAMs, altered metabolic factors, and worse RFS.
Possible viral oncolysis by members of the Reoviridae and Paramyxoviridae families (an in vitro study).
Authors: Sitkovskaya Anastasia O. , Zlatnik Elena Yu. , Kolpakov Sergey A. , Kolpakova Elena P. , Novikova Inna A. , Sagakyants Aleksandr B. , Dashkov Andrey , Kaymakchi Dmitry O. , Chupanov Gapiz M. , Gamzatova Patimat M.
Published: May 2019

e14212

Background: Our purpose was to study possible oncolytic activity of Newcastle disease virus and reovirus in different tumor cell cultures in vitro. Methods: The study was performed on LaSota strain Newcastle disease virus (NDV) vaccine (OOO NPP AVIVAK, 5000 doses) and human reovirus strains R-92 (3 serotype) deposited in the State collection of viruses of the Research Institute of Virology named after. D.I.Ivanovsky. HeLa (cervical cancer), A549 (lung adenocarcinoma), BT20 (breast cancer), PC3 (prostate cancer), HT29 (colon cancer) cell lines were used; pig embryo kidney cell culture (PEVKC) was used as a positive control. Results: After Newcastle disease virus was placed in the medium with tumor cell lines, a 5-day visual microscopic control of the cell morphology did not show signs of the cytopathic action, while a complete lysis of cell biomass was observed in the positive control (PEKC) by the third day. After the incubation with reovirus, the oncolytic activity was registered already in 24 hours: partial or complete destruction of the monolayer, decreased cell adhesion (only solitary cellular conglomerates remained at the bottom of vials), changes in the typical cell appearance. The complete cytopathogenic effect was observed after 2 days. Conclusions: The results demonstrated a marked lytic effect of reovirus in vitro, as well as varying sensitivity of the studied tumor lines to it, which may depend on their molecular genetic characteristics, for example on the RAS mutation status. Despite ambiguous results, we consider it expedient to continue the studies of these and other viruses in tumor models both in vitro and in vivo.
Reovirus strain R-92 in in vitro generation of tumor-specific T-lymphocytes.
Authors: Sitkovskaya Anastasia O. , Zlatnik Elena Yu. , Kolpakov Sergey A. , Kolpakova Elena P. , Mezhevova Irina V. , Bondarenko Elena S. , Novikova Inna A. , Dashkov Andrey , Kaymakchi Dmitry O. , Chupanov Gapiz M. , Oganyan Nairi B. , Vladimirova Liubov Yu
Published: May 2019

e14216

Background: The purpose of the study was to assess the potential of the reovirus strain R-92 for in vitro generation of tumor-specific T-lymphocytes. Methods: The human reovirus strain R-92 used as a master seed strain was previously isolated from a patient with infectious hepatitis and adapted to the growth on the pig embryo kidney cell culture (PEKC); it lost its pathogenicity after multiple passages and was characterized by electron microscopic, serological methods and PCR. HeLa lysate was prepared by the incubation with the R-92 reovirus in medium 199+L-glutamine during 5 days (37оC, 5% СО2), cytopathogenic effect was observed. Immature dendritic cells (DCs) were derived from blood monocytes cultivated for 7 days in presence of IL-4 and GM-CSF. For the DC loading during 48 hours we used: 1. HeLa lysate obtained by repeated freezing and thawing of cells (control); 2. HeLa lysate obtained by co-culturing with reovirus; 3. reovirus cultured in a standard PEKC. To assess the DC ability to activate lymphocytes, they were co-cultured for 5 days in a 3:1 ratio, 1.5x105 of target cells (HeLa) were added (1:1) for 48 hours; number of dead HeLa cells was count. At each stage of the generation of DCs and activated lymphocytes, they were phenotyped by flow cytometry. Results: DC samples 2 and 3 showed increased percentage of activated cells compared to the loaded with the control tumor lysate, CD86 and HLA-DR coexpression was 62 and 69% vs. 25%, respectively; CD83 expression was maximal after DC loading with reovirus (72.9%). After co-culturing of DC with autologous lymphocytes, the highest number of activated CD4+CD38+ and CD8+CD38+, was observed in sample 2, T-regs (CD4+CD25+CD127low) - in the control. The death of target cells cultured with activated lymphocytes obtained by DC samples 2 and 3, was 100%, control sample showed minimal cytotoxicity against HeLa. The number of T-lymphocytes in samples 2 and 3 cultured with HeLa exceeded the control values by 3-10 times. Conclusions: Reovirus R-92 used for the immature DC loading increases their presenting activity. DCs loaded with reovirus HeLa lysate, when co-cultured with lymphocytes, activate CD4+ and CD8+ and suppress T-regs, apparently leading to the total death of target tumor cells.
Correlation of radiosensitivity of lung carcinoma to CD137L endogenous expression levels.
Authors: Girdhani Swati , Rodriguez Andrew , Dua Rajiv , Parry Renate
Published: May 2019

e14229

Background: Radiation therapy is a cornerstone in the management of cancer. Preclinical and clinical evidence indicate the therapeutic efficacy of radiation therapy can be augmented by immunomodulatory antibodies against targets such as CD137 and PD-1/PD-L1. Despite evidence of synergy, the mechanistic basis has yet to be fully explored. In this study, we investigated how the expression of CD137, CD137L, PD-1, and PD-L1 correlates with radiosensitivity and proliferation of lung tumor cell lines treated with radiation as well as anti-PD-L1 and agonist anti-CD137 antibodies. Methods: Expression of CD137, CD137L and PD-L1 was measured by RT-qPCR in eight NSCLC cell lines (HCC827, HCC2935, H358, H460, A549, H596, H1650, H520) treated with radiation in doses of 0-8 Gy. Clonogenic assays for radiosensitivity and the CyQUANT assay for cell proliferation were performed after treatment with radiation in combination with anti-PD-L1 (0.01-10µg/ml) and agonist anti-CD137 (0.01-10µg/ml) antibodies. Results: Out of the eight cell lines tested, four were found to be radioresistant – H460, A549, HCC827 and H358 had SF2 (survival at 2 Gy) = 0.76, 0.70, 0.67 and 0.62, respectively – two were moderately sensitive – H520 and HCC2935 (SF2 = 0.54 and 0.40) and two were sensitive to radiation – H596 and H1650 (SF2 = 0.38 and 0.27). Expression analysis by RT-qPCR showed high endogenous expression of CD137L in three out of four radioresistant cell lines while the fourth, H358, had low endogenous expression of CD137L but upon irradiation showed more than a 3-fold induction of CD137L expression. PD-L1 expression levels measured by RT-qPCR did not correlate with radiosensitivity. In vitro cell proliferation was found to be inhibited by 48-68% in all eight NSCLC cell lines after 4 Gy radiation, but combination with anti-PD-L1 and agonist anti-CD-137 antibodies did not contribute additional toxicity. Conclusions: Our results demonstrate that CD137L expression on NSCLC cell lines correlates with radioresistance. Further, addition of anti-PD-L1 and agonist anti-CD137 antibodies to radiation showed little synergistic effect in vitro on tumor cell proliferation. Additional studies to address the synergistic effect of radiation and immunomodulation on tumor cells co-cultured with immune cells is ongoing.
Preclinical development of combination therapy targeting the dominant cytokine interleukin-1β for renal cell carcinoma.
Authors: Aggen David Henry , Ghasemzadeh Ali , Mao Wendy , Chowdhury Nivi , Chaimowitz Matthew , Hawley Jessica , Wang Vinson , Dallos Matthew , Zhang Cheng , Li Hu , Guo Xinzheng Victor , Drake Charles G.
Published: May 2019

e14237

Background: Targeting myeloid-derived suppressor cells (MDSCs), key mediators of intrinsic and adaptive immune resistance, remains a significant challenge in kidney cancer immunotherapy. One potential driver of MDSC mediated tumor immunosuppression is the cytokine interleukin-1 beta (IL-1β). Although large cancer prevention studies show that targeting IL1 decreases cancer risk, the exact mechanism of anti-IL-1 mediated anti-tumor efficacy is not yet defined. To understand the immunogenic effects of IL-1β blockade and effects on MDSCs, we used the RENCA model of RCC. Methods: We inoculated BALB/c mice with 1x106 RENCA cells. On Day +10 we treated mice (n = 8/group) with vehicle, αPD1, αIL1β, cabozantinib, combination αPD1/αIL1β, or cabozantinib/αIL1β. We harvested day +18 tumors to quantify tumor size and immune cells by flow cytometry. We then isolated immune cells for differential gene expression profiling by RNAseq. Results: Treatment with αIL1β significantly decreased tumor size compared to vehicle or αPD1 monotherapy. Combining αPD1/αIL1β or cabozantinib/αIL1β potentiated the anti-tumor effects of αPD1 or cabozantinib monotherapy respectively. Anti-IL1β therapy decreased polymorphonuclear (PMN)-MDSC infiltration (αIL1β: 0.76% +/- 0.21 vs. vehicle: 1.89% +/- 0.37, P = 0.014). Relative to PD1 monotherapy, combining αPD1 and αIL1β decreased PMN-MDSCs (αPD1: 2.14% +/- 0.56 vs αPD1/αIL1β: 0.91% +/- 0.15, P = 0.033) and increased M1-like tumor associated macrophages (TAM) (αPD1: 0.798% +/- 0.21 vs. αPD1/αIL1β: 12.04% +/- 2.9, P = 0.0008). Gene expression profiling from sorted immune cells from αIL1β treated tumors revealed multiple differentially regulated cytokines including interleukin-6, IFN-γ, and IL-10. Conclusions: Blockade of IL-1β depletes intratumoral PMN-MDSCs and increases M1-like TAM with minimal effects observed on the T cell compartment. Treatment with cabozantinib or αPD1 in combination with αIL1β led to sustained control of tumor growth. IL-1β represents a promising target for kidney cancer immunotherapy. Based on the preclinical data, a clinical trial of neoadjuvant αPD1 and αIL1β therapy is planned for patients with high risk RCC.
Analysis of the relationship between ratio N/L and survival in lung cancer patients treated with immunotherapy.
Authors: De Dios Alvarez Noemi , Costa Rivas Marinha , Agraso Busto Sara , Pereiro Corbacho Diego , Alonso Herrero Ana , García Benito Carmé , Villanueva Silva Maria Jose , Huidobro Vence Gerardo , Lázaro Quintela Martín Emilio , Castellanos Diez Javier , Casal Rubio Joaquín
Published: May 2019

e14247

Background: The neutrophil-lymphocyte (N / L) ratio is a marker of general immune response in different stress situations, having shown a relationship between the quotient and the evolution of patients treated with immunotherapy (IT), emphasizing the importance of inflammation in these patients. Methods: In order to evaluate this relationship in a context of usual clinical practice, we performed a retrospective review of patients with pulmonary neoplasia who received IT treatment in the first line or successive, between November 2015 and December 2018, excluding those who received treatment within of clinical trial. Data were collected from the clinical history of each patient, with special attention to baseline neutrophil and lymphocyte numbers, objective response to therapy by criteria iRECIST 1.1 after 3 months of treatment and overall survival (OS) defined from the beginning of treatment until death by progression of the disease. Results: 92 patients (29 women and 63 men) with a median age of 64 years (44-79) were analyzed. 15 (16,3%) patients (p) received immunotherapy as first line treatment, 65.2% (60 p) received it as 2nd line and 18,5% (17 p) as 3rd or succesive lines. The average number of cycles received was 14 (1-52). Regarding the type of IT, 54 p (58,7%) received treatment with Nivolumab, 33 patients (36%) with Pembrolizumab and 5 patients (5.3%) were treated with Atezolizumab. Two stretches of baseline N / L ratios ≤5 (low) and > 5 (high) were defined. Low ratio N / L (≤5) was identified in 62p (67.4%) of the patients treated with IT and high ratio N/L ( > 5) in 30p (32,6%). Of the 62 patients with a low ratio: 41p (66.1%) had some type of response or stabilization of their disease, 13 patients (21%) had progression and 8 patients (12.9%) received less than three months of treatment, 6 patients for PS deterioration and the other 2 patients continue with the treatment and are pending reevaluation. Among the 30 patients with high N / L quotient: 7p (23.3%) presented response or stabilization of the disease, 23 patients (76,7%) presented progression or treatment was interrupted due to deterioration of the ECOG. The average survival in the group with a low N / L ratio (≤5) was 213 weeks compared to the group with a high N / L ratio ( > 5) 144 weeks (p < 0.05). Conclusions: The N / L ratio has been identified in some studies as an adverse prognostic factor in patients treated with IT. Our data from the usual clinical practice support this theory. If these findings are confirmed in future studies, it could be used as a response biomarker for better patient selection.
Treatment outcome and safety of anti-PD-1 antibody against elderly non-small cell lung cancer.
Authors: Sakatani Toshio , Saihara Aya , Ikushima Hiroaki , Takeshima Hideyuki , Taniguchi Yuri , Hojo Masayuki , Usui Kazuhiro
Published: May 2019

e14249

Background: The anti-PD-1 antibodies, Pembrolizumab (Pem) and Nivolumab (Nivo), have become available for practical clinical use in Japan, and many patients are benefiting from them. Lung cancer is predominantly a disease of the elderly people. However, the validity and safety of anti-PD-1 antibodies against the elderly are insufficient. Methods: From February 2016 to November 2018, 66 patients with non-small cell lung cancer who received anti-PD-1 antibody therapy (23 Pem and 43 Nivo) were evaluated for progression-free survival (PFS), overall survival (OS), and safety were compared between groups between the aged ≥75 years (≥75 yrs) and < 75 years ( < 75 yrs). Results: The median age (width) was 67 years (46-87 years). 46 patients were < 75 yrs and 20 patients were ≥75 yrs. PFS was 3.3 months (m) vs. 4.4 m ( < 75 yrs vs. ≥75 yrs)(p = 0.214). OS was 11.7 m vs. 16.7 m ( < 75 yrs vs. ≥75 yrs)(p = 0.212). Similar analysis was carried out for each anti-PD-1 antibody. In Pem, 15 patients were < 75 yrs and 8 patients were ≥75 yrs, and there was no significant difference between PFS and OS (p = 0.46, p = 0.17, respectively). In Nivo, 31 patients < 75 yrs, 12 patietns > 75 yrs, and there was no significant difference in PFS and OS (p = 0.49, p = 0.44, respectively). Immunity-related adverse events (irAE) was expressed in 25 patients (37.9%). Among 66 patients, 19 in 46 (41%) were < 75 yrs, 6 in 20 (30%) were ≥75yrs. There were not many irAEs even in the elderly people (χ2= 0.757, p = 0.384). Conclusions: Pem and Nivo proved to be effective and safe for the elderly patients even in the clinical setting.
New generation of oncolytic herpesviruses embodying immunotherapeutic genes encoding IL-12 and anti-PD-1 antibody.
Authors: Zhou Grace , Ni Dongyao , Yan Runbin , Chen Xiaoqing , Liu Xianjie , Ma Jie , Tang Yuxin
Published: May 2019

e14250

Background: Systemic administration of checkpoint inhibitors alone and especially concurrent with intratumoral administration of oncolytic herpes simplex viruses (oHSV) have a major impact on cancer therapy marred by rare failures of healthy organs. Methods: We constructed 3 families of recombinant oHSVs expressing no immunomodulatory genes (T1 series), murine or human IL-12 (T2 series) and murine or human IL-12 and anti-PD-1 antibody (T3 series). We compared the oncolytic effects of a single or multiple intratumoral injection(s) of T1, T2 and T3 series oHSVs. We also compared the effectiveness of intratumoral injection of T3 oHSV with systemic administration of anti PD-1 ab and IL12 alone or in combinations with T1. Results: Insertion of the gene encoding PD-1 Ab significantly augmented the oncolytic activity of oHSV bereft of immunostimulatory genes (T1 series) or expressing IL-12 alone (T2 series). In syngeneic mice the T3 series oHSV expressing murine IL-12 and PD-1 Ab was effective against a variety of murine tumors. Concurrent with enhanced cytolytic activity the T3 oHSV induced the production and significant intratumoral accumulation of IL-12, PD-1 Ab and IFN-γ. Consistent with an earlier report we noted an inverse correlation between the volume of the tumor and the quantity of retained IFN-γ. The most effective oncolytic effect resulted from the administration of T3855 expressing both IL12 and anti PD-1 antibody or T2850 concurrently with intraperitoneal administration of anti PD-1 antibody alone. The least effective were single intraperitoneal administration of IL12 or PD-1 antibody. Intratumoral injection of T3 oHSV was most effective against murine tumors in comparison of either systemic administration of anti PD-1 ab and IL12 or intratumoral injection of T1 in combinations of systemic administration of anti PD-1 ab and IL12. Conclusions: We report the marriage in a single therapeutic agent of three distinct cancer therapies: the targeting of cancer cells by oncolytic viruses, the stimulation of immune system by IL12, and the production of immunotherapeutic antibodies against PD-1. The significant finding reported in this article is that the anti-tumor responses remain concentrated in the tumor environment.
Immunotherapy-induced endocrinopathies: A multi-center experience.
Authors: Ronan Karine , Che Othman Elly Hanis , McKenna Susan , Anderson Cian , Sheehan David , Griffin Margaret , McDermott Ray , Westrup Jennifer
Published: May 2019

e14251

Background: Endocrinopathies account for approximately 10% of immune-related adverse events.1 Methods: We carried out a retrospective review of 100 patients who received immunotherapy at two hospital sites between 2012 and 2019. Our aim was to ascertain the overall rate of immunotherapy-induced endocrinopathy and assess the impact of immunotherapy on pre-existing diabetes and thyroid dysfunction. Results: 28 out of 100 patients had pre-existing thyroid dysfunction, 36% of these experienced further thyroid dysfunction on immunotherapy. 41 patients had a diagnosis of renal cell cancer, and had treatment with a tyrosine kinase inhibitor prior to treatment with Nivolumab. Eighteen of these patients had a pre-existing thyroid dysfunction. Of these 18 patients, 44% experienced further thyroid dysfunction following immunotherapy. Of the 23 patients who received a tyrosine kinase inhibitor, who had no pre-existing thyroid dysfunction, 17% developed thyroid dysfunction on Nivolumab. 15% of patients without a pre-existing thyroid dysfunction developed a new thyroid dysfunction while on immunotherapy. Seven patients had diabetes mellitus prior to commencing immunotherapy, diabetes control did not disimprove during treatment. One case of diabetes was diagnosed on immunotherapy(1%). One case of adrenalitis(1%) and two cases of hypophysitis(2%) were diagnosed on immunotherapy. The adrenalitis occurred on Nivolumab (1.35%). Both cases of hypophysitis occurred on ipililumab(14%). The overall rate of new immunotherapy-induced endocrinopathy was 15%. Conclusions: The rates of endocrinopathy observed de novo are in line with published rates.1,2,3,4 In this small study, an increased rate of thyroid dysfunction was observed in patients with pre-existing thyroid dysfunction. Prior treatment with a tyrosine kinase inhibitor did not cause an increase in the rate of thyroid dysfunction. Ipilimumab was observed to be a risk factor for development of hypophysitis. [Table: see text]
Absolute lymphocyte count (ALC) as predictor of Pneumocystis Jiroveci Pneumonia (PCP) infection in patients on immune checkpoint inhibitors (ICPi).
Authors: Joshi Krishna Prasad , Atwal Dinesh , Siegel Eric R , Jillella Anusha , Konda Manojna , Parikh Richa , Ananthula Aneesha , Bano Kulsum , Arnaoutakis Konstantinos , Schafer Liudmila , Milojkovic Natasa , Mahmoud Fade , Govindarajan Rangaswamy
Published: May 2019

e14255

Background: Checkpoint inhibitor pneumonitis (CIP) is an uncommon but serious immune related adverse event needing treatment with prolonged steroids. PCP prophylaxis is recommended for patients on daily prednisone equivalents of 20 mg or greater for at least 1 month. Known risk factors for PCP infection in general population are glucocorticoid use and defects in cell-mediated immunity. We conducted a retrospective study to evaluate the association between ALC and development PCP infection in the patients with CIP treated with steroids. Methods: A retrospective analysis of subjects who received ICPi for any type of cancer from 2011 to 2018 at a single institution was conducted. PCP was diagnosed by PCR of bronchioalveolar lavage. In patients with CIP, ALC at the beginning of steroid treatment, dose and duration of steroids and development of PCP infection was recorded. The median ALC was compared between the patients with PCP infection and without PCP infection via Welch’s t- test at 5% alpha. Results: Out of 515 patients who received ICPi, there were 429 (83.3%) Caucasians, 79(15.3%) African Americans, 7(1.3%) others. CIP developed in 23 (4.38 %) patients. There were 15(65%) males, 8 females (35%), 20(86.9%) Caucasians and 2(8.6%) African Americans. Median age was 67 Yrs. (range 29-80). Six out of 23 (26%) patients with CIP developed PCP infection. Median ALC in the patients with PCP infection was 0.75 K/uL (range 0.2-0.83) compared to 1.14 K/uL (range 0.18 - 1.76) in those without PCP infection (p = 0.0182). Conclusions: In patients with CIP, low ALC and prolonged steroid therapy are more likely to result in PCP infection as opposed to steroid therapy alone. Low ALC and prolonged steroid therapy may select patients who require PCP prophylaxis. [Table: see text]
Defining tissue requirements for a reliable detection of tumor mutational burden (TMB) in clinical FFPE samples.
Authors: Schildhaus Hans-Ulrich , Herold Thomas , Worm Karl , Stoss Oliver
Published: May 2019

e14265

Background: In the context of immuno-oncology related cancer treatment, tumor mutational burden (TMB) is currently explored as a predictive biomarker for several human malignancies. Several sequencing assays are increasingly commercially available. Established methodologies require rather large amounts of DNA input (in the range of 100 ng) which, however, are frequently not available from small biopsies. We aim to investigate how tissue size and DNA input influence TMB scores. Methods: DNA from 20 specimens (12 biopsies of non-small cell lung cancer (NSCLC); 8 surgical resection specimens from NSCLC, colorectal cancer and endometrial carcinomas) was manually extracted by using the Qiagen GeneRead DNA FFPE kit. Cases were selected to provide a wide range of relative tumor cell content (from < 10% to > 50%) and to include microsatellite-stable and –instable (MSI) cases. Samples were analyzed in triplicates from predefined numbers of unstained sections of a standardized tissue size. DNA quantification was done fluorometrically and by qPCR. Up to 40 ng of DNA were analyzed with the QIASeq TMB Panel (incl. MSI primer boosters; Qiagen). Sequencing was done on an Illumina NextSeq platform, TMB scores and MSI status were determined by using the CLC workbench 5.0.1 (Qiagen). Results: Biopsy samples generated less numbers of DNA molecules (as detected by unique molecular identifiers, UMIs) and less overall reads compared to resection samples. UMI coverage was > 500x in all samples with > 15 ng DNA input. TMB scores were highly reproducible among all samples with > 15 ng DNA but differed significantly among samples with limited DNA input. Interestingly, TMB scores were inversely correlated with DNA input among those samples with < 15ng. Thus, valid TMB scores could also be obtained from only one slice of 1 cm2 tissue from tumor resections or 3 slices of an endoscopic biopsy (each of 5µm thickness). All pre-characterized MSI carcinomas could be detected correctly. Conclusions: We provide first evidence that TMB can be reliably determined also in small biopsies yielding limited DNA content. However, false high TMB scores can occur in samples with < 15ng DNA input. Our results contribute to the definition of minimum requirements (tissue size and DNA input) for valid TMB measurement on clinical FFPE samples.