bluuebunny/crossref_metadata_2025 huggingface dataset:
Tolerability of single agent nivolumab in cancer patients—A meta-analysis.
Authors: Huang Judy , Carvajal Alejandro Ramon , Wu Shenhong
Published: May 2020

e15146

Background: Nivolumab, a PD-1 immune checkpoint inhibitor, is being widely utilized in a number of cancers as a single agent. Adverse events, including those that are immune mediated, have been reported that resulted in discontinuation of the drug. We conducted a meta-analysis of published clinical trials to further investigate the tolerability of nivolumab in cancer patients. Methods: Databases, including PubMed and abstracts presented at American Society of Clinical Oncology annual meetings from 2015 to December 2019 were searched to identify relevant studies. This included randomized controlled trials and single arm trials that reported a discontinuation rate due to adverse events. Incidences and relative risks were calculated based on the heterogeneity of included studies. Results: Seventeen studies published between 2015 and 2019 were selected, which included a total of 4216 patients. Overall, the discontinuation rate of nivolumab due to adverse events was 6.7% (95% CI 4.7-9.5%). The rate varied significantly with the type of cancer (p < 0.001). The lowest discontinuation rate was in NSCLC of 2.1% (95% CI 1.3-2.3%), and the highest discontinuation rate was in melanoma of 15.2% (95% CI 6.8-30.6%). In addition, significantly higher intolerability is associated with the fixed dose of nivolumab at 240 mg than 3 mg/kg (P = 0.004). In comparison with chemotherapy controls, the intolerability in the nivolumab group was not significantly different, with a relative risk of 0.97 (95% CI: 0.36-2.6). Conclusions: The intolerability of nivolumab as monotherapy may be similar to chemotherapy, and can be improved with dose adjustment.
Blood-based gene expression profiling to reveal potential response biomarkers for immunotherapy in advanced lung cancer.
Authors: Wu Jie , Huang Weihua , Yin Changhong , Cheung Yee Him , Abrams Debra , Fallon John T , Dimitrova Nevenka , Fanucchi Michael Paul
Published: May 2020

e15155

Background: Novel immunotherapies are becoming a viable option for advanced lung cancer treatment. High-throughput gene expression profiling (GEP) has enabled better understanding of patient responsiveness to targeted immunotherapy. However, non-invasive blood-based signatures are needed to monitor and predict response. Methods: To evaluate the predictive and prognostic role of blood-based GEP, we designed a longitudinal study by enrolling 15 patients with advanced lung cancers. A total of 65 samples were collected for RNA sequencing, ~4 blood specimens per patient, before and during anti-PD-1 treatment. We used multiple analyses, including time-course differential gene expression, principal component, lymphocyte compartment deconvolution, and genetic mutation, to search for and assess potential predictive and prognostic aspects. Results: Of 15 patients, 11 were classified as Responders (partial responders) and four were Non-Responders (one stable and three progressive diseases). Our analyses demonstrated: 1) By comparing baseline GEPs from Responders vs. Non-Responders before the first treatment, we identified potential markers (e.g., LY6E is significantly lower expressed in Responders, with Log2 Fold change = -3.44 and p = 1.83E-04) that can be used as predictors of responsiveness of the patients; 2) Immunoglobulin subunits- and T cell receptor complex-related genes were differentially expressed in Responders (DAVID analysis, p = 6.7E-3 and 2.1E-2, respectively), but not in Non-responders; 3) γδ T cells in the lymphocyte compartment were relatively increased in Responders; 4) Despite a different set of genes differentially expressed at different time points, the biggest GEP changes were at ~ week 6, after the second treatment. Additionally, we observed certain genes consistently up- or down-regulated through the whole course of treatment. Furthermore, after the first treatment, genes in the immune response pathway were regulated to different directions in Responders and Non-Responders. For example, interleukin receptor genes, such as IL18R1 and IL18RAP, and CD24 were down-regulated in Responders, but up-regulated in Non-Responders (p = 0.042, 0.023 and 0.044 respectively, t-test for the differential expression in these two groups). Conclusions: The utility of blood GEP to identify predictive and prognostic factors for precision immunotherapy is encouraging. Nevertheless, these results, predictive of the anti-PD-1 clinical response, are preliminary and need to be validated in a larger cohort.
Correlation of immunity-related adverse events with survival and response to anti-PD-1 treatment in patients with metastatic melanoma.
Authors: Cybulska-Stopa Bozena , Lugowska Iwona , Zietek Marcin , Dawidowska Anna , Czarnecka Anna Malgorzata , Piejko Karolina , Galus Lukasz , Ziolkowska Barbara , Kieszko Stanislaw , Kosela-Paterczyk Hanna , Switaj Tomasz , Kozak Katarzyna , Rogala Pawel , Kempa-Kaminska Natasza , Calik Jacek , Teterycz Pawel , Staroslawska Elzbieta , Suwinski Rafal , Mackiewicz Jacek , Rutkowski Piotr
Published: May 2020

e15164

Background: Immunotherapy has become a standard treatment option for metastatic melanoma patients, and the use of anti-programmed cell death-1 monoclonal antibodies (anti-PD-1) has significantly improved the outcomes of this group of patients. Immune related adverse events (irAEs) during anti-PD-1 treatment may present a significant problem. However, irAEs' relationship with overall survival although suggested in single studies was not confirmed in real world data. Methods: Consecutive patients starting treatment between March 2016 and April 2019 with fist line anti-PD-1 (nivolumab or pembrolizumab) for unresectable or metastatic melanoma in 6 comprehensive cancer centers in Poland were enrolled in the study. Baseline patients characteristics and irAEs development during treatment were evaluated to identify predictors of progression-free (PFS) and overall (OS) survival in Cox model. PFS and OS were assessed using Kaplan–Meier method. Results: Overall, 410 patients were included in the present analysis and 107 patients experienced irAEs. Response rate (RR) and disease control rate (DCR) were 36% (148 pts) and 65% (267 pts) respectively. Median PFS and OS were 7.6 (2.8–21.6) and 21.6 (6.7–38.2) months, respectively. In univariate analysis normal LDH level, no brain metastases, ECOG 0 or 1, ≤ 2 number of metastases locations were positive prognostic factors for both OS and PFS. At the same time irAEs occurrence was correlated with longer PFS, OS, RR and DCR (all p < 0.001); moreover high LDH level correlated with irAEs (p = 0.027) development. There was no correlation between irAEs and the number of cycles of anti-PD-1 patients received. Conclusions: Our study showed an association between irAEs and longer survival on anti-PD1 therapy in patients with advanced or metastatic melanoma. An association with irAEs and response to therapy has also been demonstrated.
Pre-existing interstitial lung abnormalities are risk factors for immune checkpoint inhibitor-induced interstitial lung disease in non-NSCLC cancers.
Authors: Shimoji Kiyofumi , Masuda Takeshi , Nakanishi Yu , Yamaguchi Kakuhiro , Sakamoto Shinjiro , Horimasu Yasushi , Nakashima Taku , Miyamoto Shintaro , Iwamoto Hiroshi , Fujitaka Kazunori , Hamada Hironobu , Hattori Noboru
Published: May 2020

e15171

Background: Immune check point inhibitor (ICI) induced interstitial lung disease (ICI-ILD) is a clinically serious and life-threatening toxicity. Pre-existing ILD has been reported to be a risk factor for ICI-ILD in patients with non-small cell lung cancer (NSCLC). In addition, we have previously reported that interstitial lung abnormality (ILA) is also a risk factor for the ICI-ILD. Therefore, we investigated whether any patient characteristics, including ILA, were risk factors for ICI-ILD in patients with non-NSCLC cancers. Methods: Head and neck cancer, malignant melanoma, oral cavity cancer, renal cell carcinoma or gastric cancer patients who received anti PD-1 antibody (Nivolumab or Pembrolizumab) at Hiroshima University Hospital from December 2015 to May 2019 were enrolled. Information on patient characteristics before anti-PD-1 antibody administration, including chest CT findings and laboratory data, were obtained. Results: Two hundred patients were enrolled, and 20 (10%) developed ICI-ILD. Grade1 was observed in 15 patients, grade2 in 3, and grade3 and 5 in 1. There was no significant difference in the background factors between patients with and without ICI-ILD. On the other hand, the proportion of patients with ILA was significantly higher in the patients with ICI-ILD than those without (P < 0.01). Furthermore, univariate logistic regression analysis revealed ILA was the risk factor for ICI-ILD (p < 0.01), and multivariate logistic regression analysis showed that GGA or reticulation in ILA was an independent risk factor for ICI-ILD (p = 0.016, 0.011). Conclusions: Pre-existing ILA is a risk factor for ICI-ILD, and GGA or reticulation in ILA is an independent risk factor for ICI-ILD in patients with non-NSCLC cancers. Therefore, we should pay more attention to the development of ICI-ILD in patients with ILA, especially GGA or reticulation.
Lung cancer immunochemotherapy: Codelivery of EGFR tki and PTX via an immunostimulatory dual functional nanocarrier.
Authors: Diao Dingwei , Cai Kaican , Liu Xiguang , Yang Jianjun
Published: May 2020

e15177

Background: Immuno-target therapy combines a target therapy agent with an immune modulating agent and represents an attractive approach to improve cancer therapy.However, the success of it is hampered by the lack of a strategy to effectively co deliver the two therapeutics to the tumours.Here we report an improved dual functional nanocarrier (PEG5k-Fmoc-NLG2) to load Gefitinib for improved cancer Immuno target therapy.PEG5K-Fmoc-NLG2 is a prodrug that can inhibited IDO enzyme activity in vitro,while IDO1 inhibition reversed T cell suppression mediated by IDO expressing mouse cancer cells. Methods: Gefitinib loaded in carrier was prepared via a film hydrationmethod,the physicochemical properties of micelles was determined by TEM and particle size test.The cytotoxicity of the drug was detected by MTT assay. 3LL lung cancer was inoculated in mice and different drugs was injected through the tail vein to compare the anti tumor effect of drug loading micells with existing targeted drug gefitinib and CTLA4 antibody; tumor tissue proteins were extracted to clarify changes in pEGFR expressionexpression; flow cytometry will be finished to observe tumor immune microenvironment changes. Results: 1. Gefitinib loaded micelle maintain the similar size with blank PEG5k-fmoc-NLG2 micelles that is around 20nm.The drug-loaded micelles produced uniformed size and morphology. which further confirmed the size of micelles was around 20nm. 2. PTX and Gefitinib loaded in PEG5k-fmoc-NLG2 exhibited significantly enhanced tumor cell killing effect than the free drug on both A549 and 3LL cell lines. 3. The antitumor efficacy of different drugs loaded in the carrier were tested in a murine lung cancer model (3LL). All treatment showed significant better antitumor activity compared to the control group, Gefitinib/PEG5k-Fmoc-NLG2 plus CTLA4 antibody exhibited the best antitumor effect. Antitumor activity of gefitinib loaded in carrier was comparable with free gefitinib alone. 4. With various treatments the expression of p-EGFR protein level varied. Basically, compare with free gefitinib alone, the carrier loaded gefitinib was more effective in the inhibition of EGFR phosphorylation. Conclusions: We have shown that targeted delivery of PTX and Gefitinib via a immunostimulatory nanocarrier effectively improved drugs antitumor activity. More study about tumor immune microenvironment are required. Such strategy can be employed in novel therapy combining chemotherapy drugs and immunemodulating agents such as PD-1 and CTLA4.
Modulating tumor immune microenvironment by the STK11/LKB1 signaling in breast cancer.
Authors: Hsu Hui-Ping , Wang Chih-Yang , Kuo Yao-Lung , Lee Kuo-Ting , Chen Pai-Sheng , Cheung Chun Hei Antonio , Shen Che-Hung , Chang Chih-Peng , Chen Yi-Ling , Lai Ming-Derg , Shen Meng-Ru
Published: May 2020

e15185

Background: Standard treatment for breast cancer patients includes surgery, chemotherapy, radiotherapy, target and endocrine therapy. Immune checkpoint inhibitors are newly developing medications. The theoretical basis of immunotherapy is the interaction between cancer cells and tumor-infiltrating immune cells. Cancer cells secrete cytokines and create a specific tumor immune microenvironment (TIME) to attract or modulate immune cells. Further, genetic mutations or copy-number variations in cancer cells contribute to immunosuppression. Liver kinase B1 (LKB1) protein ( STK11 gene) is the upstream of AMP activated Protein Kinase (AMPK)/mammalian Target of Rapamycin Complex 1 (mTORC1) signaling pathway. STK11/LKB1 is one of the possible pathways modulating TIME. Methods: Twenty-seven breast cancer patients who developed recurrence within postoperative 24 months and 22 control cancer patients without recurrence were enrolled from National Cheng Kung University Hospital in Taiwan. Targeted deep sequencing was performed to assess the mutations among individuals with breast cancer using a panel of 143 cancer-associated genes. Bioinformatics and public databases were used to predict the protein functions of the STK11 genes. Immunohistochemical staining of LKB1 protein was performed in clinical specimens. Immune-related molecules were analyzed by RNA sequencing and cytokine array after suppression of STK11. Results: Mutations of STK11 gene were detected in recurrent patients and associated with poor prognosis of patients. From immunohistochemical study, the patients with low LKB1 expression had a worse survival. We utilized CRISPER/Cas9 system to transfect sgRNA into three mouse cell lines, including M158, NF639 and PY8119. RNA sequencing was performed in parental and Stk11-sgRNA cells. Immune-related pathways were ranked in the top 10 networks. Increased mRNA expression of Csf3 (protein G-CSF), Cxcl5, and Ccl2 was detected. The results are confirmed by cytokine array. The expression of G-CSF (gene Csf3) and CXCL5 (gene Cxcl5) proteins was increased in Stk11-sgRNA cells. The results were similar with RNA sequencing. Conclusions: Our findings suggest that suppression of STK11/LKB1 is correlated with early recurrence of breast cancer patients and contributes to modulate TIME. The STK11/LKB1 and downstream AMPK/mTORC1 pathways may be potential targets for immunotherapy.
Evidence for persistent radiation-related injury to the immune system in patients with local breast cancer.
Authors: Athale Janhavi , Broderick Kristen , Wu Xiaojun , Grossman Stuart A.
Published: May 2020

e15189

Background: Clinical data in multiple tumors has demonstrated that the inadvertent radiation of circulating lymphocytes causes grade III-IV lymphopenia which is associated with worse outcomes in cancer patients and failure to respond to immunologic interventions. Murine data from our lab demonstrated that radiation to the brain results in striking changes in the anatomy and cellularity of distant unirradiated lymph nodes. This study was designed to understand the relationship between local radiation and the depletion of distant unirradiated lymph nodes in humans which has not been previously studied. Methods: Adult women with breast cancer who had undergone prior mastectomy with pathology, labs, and radiation data available at our institution were enrolled at the time of their deep inferior epigastric perforator artery (DIEP) flap reconstruction. During reconstruction, a single abdominal lymph node was extracted, and subsequently formalin fixed. The pre- and post-treatment lymph nodes of radiated and non-radiated patients were presented in a blinded manner to the hematopathologist. The pathologist described each lymph node and graded the lymph node density as (1) low, (2) low-normal, (3) normal, or (4) high. Results: Seven women have been enrolled (median age 50; range 31-55) with AJCC tumor stages from 1a – 3b (five are hormone positive, and two are triple negative). The reconstruction was completed on average 488.71 + 224.57 days after initial mastectomy. Three of the women received neoadjuvant chemotherapy, and five received adjuvant chemotherapy. Five of the seven women had received radiation (mean 50.9 + 5.6 Gy). The baseline median LN density score in all patients was 4 (range 1-4), with a post mean LN density of 1.2 + 0.4 in the radiation group compared to a mean score of 3 + 1.4 in the control group. Conclusions: Systemic lymphocyte counts remain relatively stable but lymph node density is markedly lower than baseline in 80% of patients who received chest wall irradiation consistent with data from animal model studies. It is important to note that this effect on the nodes is seen long after the radiation has been completed. [Table: see text]
Tumour immune characterization in adenoid cystic carcinoma identifies prognostic and immunotherapeutically relevant messenger RNA signatures.
Authors: Dou Shengjin , Chen Di , Li Rongrong , Yang Yining , He Ning , Jin Ge , Li Jiang , Zhu Guopei
Published: May 2020

e15191

Background: Here, we profiled genetic hallmarks of Chinese adenoid cystic carcinoma (ACC) patients and immune cells infiltration levels by gene expression-based computational methods to investigate the correlation with immunotherapy. Methods: 54 patients with pathologically confirmed ACC admitted from January 2016 to October 2019 were included. Mutation and expression profiles were obtained by whole-exome sequencing (WES) and RNA-seq, respectively. A nomogram that integrated clinicopathological features with the immune signature to predict recurrence or distant metastasis probability was constructed by multivariate Cox regression. Results: As expected, we observed MYB fusion and somatic mutations in our cohort (57%, 31/54), strengthening the role of these aberrations as critical events in ACC. Mutations in NOTCH1 were also found in our cohort (19%, 10/54). Median tumour mutational burden (TMB) of these patients is 0.85 Muts/Mbp (0-11 Muts/Mbp). Analysis of WES data showed that all patients were microsatellite instability (MSI)-negative, and 17 of them were confirmed by conventional MSI-PCR testing. Lack of PD-L1 expression is 96.0% (n = 46/48) at a tumour proportion score 0% and 22.9% (11/48) of cases showed more than 10% CD8+ prevalence. We compared cancer types using an immune infiltration score (IIS) and an antigen presenting machinery expression (APM) score and find that ACC is among the lowest for both scores. Statistically significant correlations were found between IIS and distant metastasis-free survival (univariate analysis, HR = 2.64, 95% confidence interval [CI], 1.134 to 6.145, p = 0.023). Patients with higher IIS had shorter distant metastasis-free survival time (log-rank p = 0.020), and patients with higher T cell infiltration score (TIS) had shorter locoregional recurrence-free survival time (log-rank p = 0.050). In addition, we investigated the pretreatment immune profile of five ACC patients treated with PD-1 checkpoint inhibitors, and found that both TIS and APM were elevated in responding patient (with a partial response to camrelizumab) whereas they were in the lowest for patients with progressive disease on camrelizumab or pembrolizumab. Conclusions: Our study highlights the immune infiltration patterns and unravels mRNA signatures with prognostic utility and immunotherapeutic biomarker potential in ACC.
A novel TGF-beta targeted peptide HTPEP-001 enhances antitumor efficacy of anti-PD1 in mice models.
Authors: Li Xiaomei , Ding Yi , Xiao Ling , Chen Rui , Qing Ying , Jiang Qiu , Gao Yuanxue
Published: May 2020

e15200

Background: With the emergence of immunotherapy in recent years, therapeutic antibodies against programmed death 1 (PD1) and its ligand (PDL1) have proven to be a huge success in multiple cancer types. However, these robust responses only occur in a subset of patients. Identifying mechanisms of responses and resistance is key to improve outcomes and setup new treatment strategies. Transforming Growth Factor-beta (TGF-beta) is an immunosuppressive cytokine and overexpression of TGF-beta in the tumor microenvironment is proven to be a key element of resistance to anti-tumor response. We have developed a novel peptide HTPEP-001 which blocks TGF-beta signaling in both cellular and fibrosis animal models (ATS-2020 abstract). In this study, we explored the antitumor effect and mechanism of HTPEP-001 when administered together with anti-PD1 antibody in mice models. Methods: Animal models: 1. C57BL/6 mice were inoculated subcutaneously with 5*105 MC38 cells. 2. BALB/c mice were inoculated on the right mammary fat pad with 1*105 EMT-6 cells. Treatment groups: A. HTPEP-001 (60 mg/kg, intravenously, qd); B. isotype IgG; C. rat anti-mouse PD1 (10 mg/kg, intraperitoneally, twice a week); D. Coadminstration of HTPEP-001 and anti-PD1 (combo). Treatments were initiated 4-5 days after inoculation and lasted for 10 days. ELISA, immunofluorescence and flow cytometry analysis of dissected tumors were performed. Results: Compared to anti-PD1 group, mice treated with both HTPEP-001 and anti-PD1 exhibited additional reductions in tumor size and 13% higher inhibition of tumor growth (59% in Anti-PD-1 vs 72% in combo) in MC38 model. In the EMT6 model, coadministration of HTPEP-001 and anti-PD1 also showed additional reduction in tumor burdens. Coadmistration of HTPEPE-001 and anti-PD-1 significantly decreased active TGF-beta1 and phosphorylated Smads expression in tumors in both MC38 and EMT6 models. Furthermore, immunofluorescence and flow cytometry analysis of CD8+ lymphocytes in tumor mass revealed markedly elevated number of cytotoxic tumor infiltrating lymphocytes in combo group compared to HTPEP-001 and anti-PD1 groups. Conclusions: This study showed that HTPEP-001 enhanced the anti-tumor activity of anti-PD1 via suppressing TGF-beta signaling and promoting infiltration of cytotoxic lymphocytes in two syngeneic tumor models. We will continue to develop HTPEP-001 as an add-on therapy to checkpoint inhibitors with a goal to improve the outcomes of current immunotherapy.
Intratumoral use of self-delivering RNAi to reprogram the tumor microenvironment and boost the antitumor response.
Authors: Dispersyn Gerrit , Lakshmikanthan Sribalaj , Yan Dingxue , Jedinak Andrej , Maxwell Melissa , Kadiyala Mani , Cardia James , Fricker Simon P
Published: May 2020

e15206

Background: Immunotherapy targeting immune checkpoints has shown significant benefit in many cancers. However, systemic delivery can result in severe auto-immune toxicities, the risk increasing with the combination of checkpoint inhibiting antibodies. Local delivery of immunotherapies allows for multiple combination therapies, while potentially preventing significant systemic toxicities. One approach is to use self-delivering RNAi technology, INTASYL™, which has been shown to be safe and effective in clinical applications following local administration. Here we show the use of the INTASYL platform as a checkpoint inhibiting immunotherapy for intratumoral applications, through a series of preclinical in vivo studies. Methods: INTASYL compounds were made against mouse PD-1, TIGIT, PD-L1 and BRD4. INTASYL comprises an asymmetric duplex, a small duplex region (≤ 15 base pairs), a single-stranded phosphorothioate tail and chemical modifications to confer stability resulting in efficient cellular uptake allowing in situ delivery. The efficacy and immunomodulation of these INTASYL compounds was evaluated in the Hepa1-6 and CT26 murine syngeneic tumor models following intratumoral administration. Tumor volumes were recorded throughout the study. Tumor-Infiltrating-Lymphocytes (TILs) were isolated twenty-four hours post the last dose and examined for gene silencing by RT-qPCR. Tumor microenvironment (TME) and T cell activation were analyzed by flow cytometry. Results: Results showed dose-dependent attenuated tumor growth compared to control groups. Relevant changes in the TME included an increase of CD8+ T cells and an increase of activation markers on TILs. Tumor growth reduction correlated with silencing of target mRNA and immune effector cell levels and activation. Conclusions: These studies demonstrate the feasibility of using INTASYL compounds intratumorally to overcome the immunosuppressive TME, resulting in changes in T cell composition and activation, and inhibition of tumor growth. Together, these novel findings support using intratumoral INTASYL as a viable approach to immunotherapy and warrants further investigation in patients.
Modulation of CT-genes transcriptional activity in HT-29 cells by irradiation with a linear accelerator.
Authors: Kutilin Denis S. , Kosheleva Natalia G , Gusareva Marina A. , Legostaev Vladislav M. , Shlyakhova Olga V. , Dontsov Vladimir A. , Gaziev Umar Muhmadovich , Udalenkova Irina A. , Rozenko Lyudmila Y. , Donskaya Aliya K. , Rodionova Olga G. , Fatkina Natalya B. , Kit Oleg Ivanovich
Published: May 2020

e15218

Background: Cancer-testis antigens (CTAs) are used as targets in immunotherapeutic approaches based on gene-cell vaccines (GCV). In some cases, it is possible to use experimental combinatorial therapy regimens that combine radiation and GCV. Despite numerous studies of the expression of CTA in tumors of various locations, including colon tumors, the transcriptional activity of CT-genes in tumor tissue of the colon after radiation therapy remains poorly studied. The aim of the study was to analyze changes in the expression of CT-genes in HT-29 cells after irradiation with a linear accelerator. Methods: The study used human cell culture HT-29. For the model experiment, 7 Gy fractionation option was used (irradiation was carried out 5 times every 24 hours). Irradiation was performed on a Novalis TX linear accelerator (Varian, USA). The ratio of living and dead cells was calculated in a Goryaev chamber using a 0.4% trypan blue dye solution. After microscopy on the 5th day of irradiation, the cell line was removed from the vial substrate by Trypsin/Versen solution. RNA was isolated by the Chomczynski&Sacchi method. For cDNA synthesis a set of "REVERTA-L" reagents used. Expression of 16 CT genes (MAGE-A1, -A2, -A3, -A4, MAGE-B1, -B2, GAGE-1, -3, -4, MAGEC1, BAGE, XAGE3, NYESO1, SSX2, SCP1, PRAME1) was determined using the Real-Time qPCR method (reference genes — GAPDH and B2M). Statistical analysis was performed using univariate analysis of variance (ANOVA) with repeated measurements. Results: After 5 days of exposure at a dose of 7 Gy, only 20% of the initial number of HT-29 cells remained viable. It was found that irradiation was statistically significantly (p < 0.05) influenced the increase in the expression of genes MAGEA1 by 1.8 times, MAGEB1 by 6.8 times, BAGE by 1.6 times, CTAGE1 by 3.3 times and SSX2 by 5.8 times relative to intact cells. Conclusions: Thus, tumor cells exposed to radiation at a dose of 7 Gy show an increase in the expression of 5 CT-genes, which, in turn, can increase their immunogenic potential.
Intratumor immunotherapy utilizing real time radiological image guidance: Early experience from a tertiary cancer center.
Authors: Murthy Ravi , Sheth Rahul , Subbiah Vivek , Janku Filip , Naing Aung , Rodon Jordi , Yap Timothy A , Diab Adi , Patel Sapna Pradyuman , Tam Alda , Gupta Sanjay , Meric-Bernstam Funda
Published: May 2020

e15223

Background: Intratumoral (IT) delivery of immunotherapeutic agents is a compelling approach to overcoming the systemic barriers of toxicity and lack of efficacy associated with many novel agents. In this context, the application of IT delivery is growing rapidly, however the feasibility and safety profile of these interventions for lesions that require real time radiological image-guidance remains unknown. Methods: Patients who underwent IT injections of immunotherapeutic agents across several clinical trials over a 5 year period at a single tertiary cancer center were included in this analysis. Technical success & procedure related adverse events of the IT injection and the frequently accompanying concomitant or sequential image-guided biopsy of injected (adscopal) and non-injected (abscopal) sites were analyzed. Results: A total of 256 patients with metastatic &/or unresectable solid malignancies underwent a total of 1096 (median 5) image-guided IT investigational agent injections during the study period. There were no adverse events attributable to the technical component of the procedure, specifically during the needle insertions or 3862 concomitant biopsies. Soft tissue and nodal lesions in the extraparietal locations & visceral lesions in deeper, intraparietal locations and solid organs were also injected (adrenal, liver and lung). The median injected lesion tumor length was 3.3cm. Serious adverse events (NCI CTC AE ≥ 3) including dyspnea and severe flu-like symptoms developing within 24 hours of the injection & required hospitalization following 1.5% of injections. Conclusions: Intratumoral immunotherapeutic injections & tissue sampling with real time radiological image guidance are feasible across a wide range of regions and organs irrespective of agent and histology. Post-delivery anticipated & adverse events were rare & manageable.
Re-introducing immunotherapy in patients surviving immune checkpoint inhibitors-mediated myocarditis.
Authors: Peleg Hasson Shira , laufer-Perl Michal , Wolf Ido , Sivan Ayelet
Published: May 2020

e15228

Background: Immune checkpoint inhibitors (ICI) have transformed the standard care in cancer treatment. Recent case reports describe ICI-mediated myocarditis with an atypical presentation and fatal potential which lead to permanent interruption of immunotherapy. We aim to characterize ICI-mediated myocarditis and re-introduction to immunotherapy. Methods: We retrospectively evaluated the presentation, severity, and prognosis of patients diagnosed with ICI-mediated myocarditis during 2019, and presented the clinical course and outcomes of patients that were chosen for re-introduction. Results: Among seven patients, only one patient had a history of cardiac disease. The majority were diagnosed with lung adenocarcinoma and treated with anti-programmed death-1 antibody (57%). All patients were treated with single agent ICI. Most patients presented with cardiac symptoms, elevated troponin and typical magnetic resonance imaging; however only 43% had reduced ejection fraction. Five patients were defined as grade I-II and two as grade III-IV. Overall, three patients were chosen for re-introduction with concomitant low dose steroids and weekly troponin follow-up. Two patients diagnosed with grade I/II renewed therapy successfully with no recurrence of symptoms and improvement in disease burden, while the one patient diagnosed with grade III developed worsening of cardiac symptoms after the 1st cycle and therefore therapy was interrupted permanently. Overall, survival was higher among the re-introduction patients (67% vs. 25%). Conclusions: ICI-mediated myocarditis is potentially fatal and leads to permanent interruption of life saving cancer therapy. We imply that re-introduction may be considered in low grade patients; however, better definition of the diagnosis and grading is needed.
Impact of corticosteroids use on efficacy of immune checkpoint inhibitors in cancer patients: A meta-analysis.
Authors: Li Jiarui , Yang Kaili , Zhao Lin , Bai Chunmei , Sun Zhao
Published: May 2020

e15234

Background: Corticosteroids are commonly used for management of cancer-related symptoms or immune-related adverse events (irAEs) in cancer patients treated with immune checkpoint inhibitors (ICIs). However, given the inhibitory effects of corticosteroids on a broad range of immune responses, it is presumed that the use of these agents may affect the clinical outcomes of ICIs. This meta-analysis aims to explore the impact of corticosteroids use on the efficacy of ICIs in cancer patients. Methods: We conducted a search covering electronic databases (MEDLINE, EMBASE, and CENTRAL), conference abstracts (ASCO and ESMO) and reference lists to identify relevant studies. Studies that reported clinical outcomes of patients with corticosteroids administration before and/or after the initiation of ICIs treatment were eligible for evaluation. The primary outcomes included progression-free survival (PFS) and overall survival (OS). The random-effects model was utilized to synthesize the effect sizes of individual studies. Results: The initial literature search identified 1,900 records. After study selection, a total of 15 studies with 14,123 patients were included in the quantitative analysis. Corticosteroids use significantly reduced PFS (HR = 1.84; 95% CI: 1.30–2.61; P = 0.001) and OS (HR = 1.41; 95% CI: 1.18–1.68; P < 0.001) in cancer patients treated with ICIs. In subgroup analysis, corticosteroids use for cancer-related symptoms was associated with a shorter PFS (HR = 1.98; 95% CI: 1.40–2.78; P < 0.001) and OS (HR = 1.88; 95% CI: 1.25–2.83; P = 0.003), but their use for irAEs did not show a detrimental impact on OS (HR = 1.05; 95% CI: 0.77–1.44; P = 0.740). Conclusions: This meta-analysis indicated that corticosteroids use might hinder the efficacy of ICIs in cancer patients. The indications of corticosteroids use should be strictly controlled during the course of immunotherapy.
Immunohistochemical determination of PD-L1 expression with three different antibody clones and determination of SOX10 expression in triple negative breast cancer.
Authors: Guhl Margit Maria , Bohle R.M. , Ertz Martin , Kasoha Mariz , Hammadeh Mohammed Eid , Juhasz-Boess Ingolf , Solomayer Erich-Franz
Published: May 2020

e15237

Background: In triple negative breast cancer (TBNC), checkpoint inhibitors directed against PD-L1/PD-1 show an improvement of therapy. For the immunohistochemical diagnosis of the predictive biomarker PD-L1, there are various antibodies and kits available. We examined the expression of PD-L1 in TNBC with different antibody clones to compare these results with regard to the staining of tumor cell membranes, staining of immune cells and cytoplasmic staining in order to find out whether the different clones or methods can be exchanged. It was also checked whether PD-L1 or SOX10 expression correlates with the existing clinical parameters. Methods: Breast cancer tissue of 60 patients with TNBC were examined for the expression of PD-L1 and SOX10 by immunohistochemistry. The detection kit used, was the ultraView universal alkaline phosphatase red detection kit for the antibodies anti-human PD-L1 clone 22C3 from Dako, the clone 28-8 from abcam and the SOX10 antibody clone SP267 from Cell Marque. The anti-PD-L1 antibody clone SP142 was detected with OptiView DAB. The cut-offs for the expression of PD-L1 at the tumor cell membrane were < 1%, > 1 to < 50% and > 50%. In the evaluation of SP142 the staining of the immune cells was evaluated with the score: percentage of PD-L1 positive immune cells to the tumor cells that were present. For SOX10, the nuclear staining was evaluated with the score: < 1, > 1 to 50, > 50 to > 100 and 100. The relationship between PD-L1 expression (TC and IC) and SOX10 expression was evaluated with the clinical parameters of the patients from the time of diagnosis until the end of data collection. Results: The antibodies clone 22C3 and clone 28-8 lead to the same results (22,0 % PD-L1 (TC) positive). Clone SP142 showed significantly (p < 0,001) more PD-L1 positive cases(40,7%). For the additional evaluation of the cytoplasmic staining with clone 22C3 and clone 28-8 it could be shown that the PD-L1 Expression is equivalent (40,7%) to the immune cell staining with clone SP142. With exception of Ki67 we were unable to demonstrate any correlation between PD-L1 (membrane and cytoplasmic), SOX10 and other clinical parameters in TBNC. Conclusions: The antibodies clone 22C3 and 28-8 can be used equivalently for PD-L1 determination in TBNC. Clone SP142 showed different results. The cytoplasmatic staining with 22C3 and 28-8 could gain importance in the future because the results were equivalent to the immune cell staining with clone SP142. There is no correlation between PD-L1 and SOX10 in TNBC.
Tumor lymphocytic infiltration impacts recurrence in endometrioid-type endometrial carcinoma.
Authors: Avila Monica , Fellman Bryan M. , Broaddus Russell
Published: May 2020

e15239

Background: Endometrial cancers that have mismatch repair deficiency are associated with higher numbers of tumor-associated lymphocytes, but the clinical significance of this observation is unknown. Our objective was to quantify CD3+ and CD8+ tumor lymphocytes of MMR intact (MMRi) and MMRd endometrioid-type endometrial carcinomas and determine if there was an association with survival. Methods: MMRd was defined as endometrial carcinomas with loss of MLH1 expression due to MLH1 gene methylation and determined by immunohistochemistry. MMRi was defined as positive expression of MLH1, MSH2, MSH6, and PMS2. This was followed by Aperio image-based quantification was used to assess CD3+ and CD8+ lymphocyte populations in different regions of the primary endometrial carcinomas, including tumor periphery (tumor-myometrial interface), tumor center (bounded on all sides by tumor), and tumor hotspot (area with highest number of lymphocytes). Recurrence-free survival was estimated using Kaplan Meier and Cox regression. Median follow up time was 44 months. Results: 180 patients with endometrial carcinoma were analyzed of which 132 were MMRi and 48MMRd. The MMRd group had significantly higher levels of CD3+ and CD8+ lymphocytes regardless of which tumor region was assessed (Figure 1a, P < 0.001). Lymphocyte counts in both MMRd and MMRi groups had wide standard deviations such that there was some overlap in counts between the groups (Figure 1). Both MMRd and higher CD3+ counts were associated with worse recurrence-free survival. CD3+ quantification in the tumor periphery captured 21/23 recurrences (Figure 1b, HR = 8.04; 95% CI: 1.88 -34.31; p = 0.005); this included all of the MMRd cases that recurred and 7 MMRi cases with higher numbers of CD3+ lymphocytes that also recurred. Conclusions: MMRd endometrial cancers have increased numbers of CD3+ lymphocytic infiltrates within the primary tumor. Higher CD3+ infiltration is associated with greater risk of recurrence regardless of tumor location. In predicting tumor recurrence, lymphocytic counts performed better than assessment of MMR. Thus, quantification of CD3+ lymphocytes should be explored as a predictive biomarker.
Effect of CCND1 amplification on immunosuppression and the association with a poor prognosis to immune checkpoint inhibitors in solid tumors.
Authors: Huang Yingying , Chen Chuan-ben , Chen Yu , Gao Xuan , Li Yi , Lin Jing , Guan Yan-Fang , Xia Xuefeng
Published: May 2020

e15249

Background: CCND1 amplification relevant to malignant biological behavior exist in solid tumors. The prevalence and utility of CCND1 amplification as a biomarker for the clinical response to immune checkpoint inhibitors (ICIs) therapy is unknown. Methods: This study included three cohorts: Geneplus Institute ( n= 6536), The Cancer Genome Atlas ( n= 10562) and Memorial Sloan Kettering Cancer Center ( n= 106109). Comprehensive profiling was performed to determine the prevalence of CCND1 amplification and their correlation with the prognosis and the response to ICIs. Results: CCND1 amplification occurs in many cancer types, correlates with shorter overall survival and inferior outcomes with ICIs therapy. Transcriptomic analysis showed various degrees of immune cells exclusion, including cytotoxic cells, T cells, CD8+ T cells, DC cells, B cells in the tumor microenvironment (TME) in a CCND1 amplification population. The gene set enrichment analysis suggested that CCND1 amplification correlates with multiple aggressive, immunosuppressive hallmarks including epithelial mesenchymal transition, TGF-β signaling, KRAS signaling, PI3K/AKT/mTOR signaling, p53 pathway and hypoxia signaling in solid tumors. Conclusions: Our study indicated that CCND1 amplification may be a key point related to immunosuppression in TME and multiple malignancy hallmarks, and it hinders not only the natural host immune responses but also the efficacy of ICIs in solid tumors.
Prevalence of prkdc mutations and association with response to immune checkpoint inhibitors in solid tumors.
Authors: Li Yi , Chen Chuan-ben , Chen Yu , Guan Yan-Fang , Huang Yingying , Lin Jing , Chen Gang , Xia Xuefeng , Chang Lianpeng , Yi Xin
Published: May 2020

e15250

Background: Predictive biomarkers of response to immune checkpoint inhibitors (ICIs) help to identify cancer patients who will benefit from immunotherapy. PRKDC is an important gene relates to DNA double-strand break (DSB) repair and central T-cell tolerance. We aim to investigate the association between PRKDC mutations and tumor mutation burden (TMB), tumor microenvironments (TME), and response to ICIs. Methods: The whole exome sequencing data of 4022 solid tumor samples from the Cancer Genome Atlas (TCGA) and the panel-based sequencing data of 4652 solid tumor samples from GenePlus-Beijing, China were used to analyze TMB. The mRNA expression data of 3541 solid tumor samples from TCGA was used to explore the effect of PRKDC mutation on TME. Four ICIs-treated cohorts were analyzed for verifying the correlation between PRKDC mutation and the response to ICIs. Results: In TCGA datasets and GenePlus datasets, we both found that the TMB in PRKDC mutation samples were significantly higher than PRKDC wild-type samples(P < 0.05, P<0.0001 respectively). When compare to other pivotal DNA damage response (DDR) genes, it showed an identical effect of the degree in TMB by PRKDC mutation with MMR ( PMS2/ MLH1/ MSH2/ MSH6) genes, POLE/D1 and BRCA1/2 mutation (P > 0.05). Further, in TCGA datasets, it showed that PRKDC mutation samples were associated with significant expression increase of CD8+ T cells, NK cells, immune-checkpoint, chemokines etc compared to PRKDC wild-type samples (P < 0.05). In ICIs-treated cohorts, we also found the PRKDC mutation was associated with superior survival(median PFS, not reached (NR) vs. 6.8 months HR, 0.2893 95% CI, 0.1255-0.6672 P = 0.0650, Hellmann cohort; median OS, 1184 days vs. 250 days HR, 0.5126 95% CI, 0.2715-0.9679 P = 0.1020, Allen cohort), and the superiority was significantly in multivariate Cox regression analyses (HR, 0.361 95% CI,0.155-0.841; P = 0.018, Allen cohort; HR, 0.240 95% CI,0.058-0.998; P = 0.050, Hellmann cohort). Conclusions: PRKDC mutations are associated with an increased TMB, an inflamed TME and a better response to ICIs, it may be a potential predictive biomarker for ICIs-immunotherapy.
Technical validation of PD-L1 SP142 assay for TNBC in a large academic center: Unexpected challenges.
Authors: Nofech-Mozes Sharon , Slodkowska Elzbieta
Published: May 2020

e15253

Background: The IMpassion 130 trial reported that 41-46% of TNBC are positive for PD-L1 SP142 based on centralized testing. While the FDA approved it as a companion diagnostic assay with cut off value of 1%, there are no published guidelines for technical validation in local laboratories. Furthermore, with the need to evaluate staining only in inflammatory cells (IC) the readout becomes particularly challenging for cases around the cutoff point. We report our prevalence of PD-L1 positivity and proportion of borderline cases in our validation. Methods: We ran PDL1 SP142 using the recommended protocol from Roche on the following groups: G1 - 53 consecutive TNBC cases (excisions tumors ≥7mm), G2 - 10 consecutive TNBC cases (breast specimens only) that would have met the inclusion criteria for the trial (LABC with poor NAT response or metastatic), G3 – proficiency testing slide from an EQA provider (4 cores of TNBC 1 negative, 1 borderline low positive and 2 high positive). All cases were read by 2 breast pathologists that received training on PDL1 readout by Roche (one – in class and online, one – online). Results: Borderline staining around the 1% cutoff (11 of 63) and low positive (9 of 63) were encountered in the validation sets more often than shown in training sessions. Classification of borderline cases may have a substantial impact on the rate of PDL1 positivity that potentially could approach 75%. Borderline cases often have IC staining either at the advancing tumor edge, as small collections of highly positive IC or around benign ducts entrapped within the tumor. Conclusions: The prevalence of PDL1 positive TNBC in all comers could be higher than observed in IMpassion 130 (at least 50-60%). Pathologist training in readout is of utmost importance due to high rate of borderline/low positive cases raising the need for central testing in borderline cases. Guideline recommendations for technical validation similar to other class II biomarkers would be useful. Studies on interobserver variability are needed to further address this potential issue. [Table: see text]
Predicting tumor mutational burden in head and neck squamous cell carcinoma based on CT imaging features: A TCGA/TCIA study.
Authors: Luo Yangkun , Li Lu , Yin Gang , Lang Jin Yi
Published: May 2020

e15254

Background: Immunotherapy has substantially changed the therapeutic strategies for cancers. Unfortunately, only 20–50% of patients with advanced solid tumours respond to treatment. There is therefore a need for the development of methods to identify patients who are most likely to respond to immunotherapy. Tumor mutation burden (TMB) have been served as the most prevalent biomarkers to predict immunotherapy response. This study was designed to investigate the ability of radiomics to predict TMB status in patients with head and neck squamous cell carcinoma (HNSCC). Methods: TMB values were calculated using genomic data obtained from the HNSCC dataset in The Cancer Genome Atlas (TCGA).We identified matching patients (n = 100) who underwent contrast-enhanced CT scan prior to treatment from The Cancer Imaging Archive (TCIA),and patients were grouped based on the cutoff value; high group(>4.2 mutations/Mb) and low group(≤4.2 mutations/Mb). A total of 249 radiomics features(9 non-texture features and 240 scan-texture-parameter features) were extracted from CT images of the tumor. The incorporation of features into multivariable models was performed using logistic regression. The multivariable modeling strategy involved imbalance-adjusted bootstrap resampling in the following four steps leading to final prediction model construction: (1) feature set reduction; (2) feature selection; (3) prediction performance estimation; and (4) computation of model coefficients. The performance was evaluated in terms of area under the curve (AUC), sensitivity, and specificity. Results: Among all the features, twenty features were found to have the most impact on the predictive value; the two top texture parameters were GLCM-Variance and GLCM-Sum Average. In multivariable analysis, the best performance was obtained using a combination of seven texture features that can discriminate between high mutation burden versus low mutation burden. The AUC, sensitivity, and specificity of this model were 0.97 ± 0.01, 0.92 ± 0.04, and 0.92 ±0.01, respectively. Conclusions: The proposed CT-derived predictive model can accurately predict TMB status in patients with HNSCC. It may be helpful in guiding immunotherapy in clinical practice and deserves further analysis.