bluuebunny/crossref_metadata_2025 huggingface dataset:
Combined treatment of patients with bone metastases from various cancers with nivolumab plus denosumab: A retrospective study.
Authors: Ozaki Yukinori , Miura Yuji , Yamanaka Taro , Takemura Kohji , Kizawa Rika , Takahashi Momoko , Masuda Jun , Kondoh Chihiro , Tanabe Yuko , Takano Toshimi
Published: May 2019

e14153

Background: Monoclonal antibodies against the programmed cell death protein-1 (PD-1), e.g., nivolumab, have been shown to exhibit antitumor immunity and improve survival in patients with various cancers. Recently, it was reported that bone metastases are unlikely to respond to immunotherapy, while other reports have suggested that synergistic antitumor effect can be obtained by administration of denosumab in combination with PD-1 blockade using nivolumab. However, there are still very limited data on the efficacy and effects on the prognosis of nivolumab administered in combination with denosumab in patients with bone metastases. Methods: We reviewed the medical records and pharmacy database of patients who had received nivolumab plus denosumab treatment from April 2011 to March 2018 at our institution, in order to analyze the prognosis of patients with bone metastases from various cancers. The primary objective was to compare the progression-free survival between patients with and without bone metastases. The impact of combined nivolumab plus denosumab therapy was evaluated as an exploratory endpoint. Results: We extracted 201 patients who had received denosumab for bone metastasis and 45 patents who had received immune checkpoint inhibitor. We analyzed the data of 39 patients who had received nivolumab treatment; of these 39 patients, 10 had additionally also received denosumab for the treatment of bone metastases. The median age of the patients was 65 years (range 41-87); the primary disease was renal cell carcinoma in 17 patients, gastric cancer in 9 patients, melanoma in 6 patients, non-small cell lung cancer in 4 patients, and head and neck cancer in 3 patients. The median progression-free survival following treatment with nivolumab was 2.1 months in the group with bone metastases and 3.5 months in the group without bone metastases (log-rank test; p = 0.0779); the 6-month progression-free survival rate was 26.7% (95% CI: 21.6-69.2) in the patients with bone metastases and 39.3% (95% CI: 10.4-53.3) in those without bone metastases. Concurrent administration of denosumab, age, and the primary disease had no influence on the progression-free survival rate. Conclusions: This retrospective analysis revealed a trend towards lower immune responses in patients with bone metastases, even when nivolumab is administered in combination with denosumab. Immunotherapy for bone metastasis needs to be further explored.
Preclinical characterization and efficacy of 4R9, a novel immune checkpoint blockade targeting CEACAM1 for cancer therapy.
Authors: Lee Jae-Chul , Hur Minkyu , Park Hye-Young , Lee EunHee , Yum Hye In , Nam Hye-mi , Oh Mi-Young , Choi HyungSuk , Kim Jae-Hwan , Cho Byoung Chul , Lim Yang-Mi , Won Jonghwa
Published: May 2019

e14155

Background: CEACAM1 is one of the several immune checkpoint receptors expressed on T cells that mediate suppression of inflammatory T cell response. It is known that CEACAM1-CEACAM1 homophilic interaction induces downregulation of ZAP70 phosphorylation in response to T cell receptor (TCR) stimulation. Also, CEACAM1 is highly expressed on non-small cell lung cancer (NSCLC) and its expression correlated with cancer progression and poor prognosis. We developed a fully human monoclonal antibody 4R9, human CEACAM1-targeting antibody. Methods: T cell activation of 4R9 was determined by NFAT-luciferase reporter assay with CEACAM1 overexpressing Jurkat stable cells. In vitro efficacy of 4R9 was examined using NK-mediated tumor cell killing assay. The anti-tumor efficacy of 4R9 alone or in combination was studied in vivo in a humanized mouse model engrafted with NSCLC patient-derived tumor xenografts. Results: Anti-CEACAM1 antibody 4R9 binds to CEACAM1 overexpressed in HEK293 or Jurkat cells but not to other CEA family members. 4R9 blocks CEACAM1-CEACAM1 homophilic interaction by binding to N domain of CEACAM1. CEACAM1-CEACAM1 homophilic interaction induced downregulation of ZAP70 phosphorylation in response to TCR stimulation in CEACAM1 overexpressing Jurkat stable cell line, which was rescued by 4R9 resulting in augmentation of NFAT activity and IL-2 expression. NK cell-mediated tumor lysis was increased by 4R9 in a CEACAM1 expression-dependent manner. Out of 49 NSCLC tumor tissues, 20 cases exhibited dominant expression of CEACAM1 over PD-L1 with 6 cases showing > 50% of CEACAM1 positivity. In a single mouse trial using NSCLC PDX-huNSG mouse model, 4R9 (20 mpk, 2qW) suppressed tumor progression more than 30% as monotherapy (10/19) as well as in combination (16/22) with pembrolizumab (5 mpk, 2qW). Moreover, PDX of adenocarcinoma origin with more than 50% of CEACAM1 expression were more efficiently prohibited for progression with 4R9, suggesting potential therapeutic use of 4R9 in patients with NSCLC. Conclusions: Anti-CEACAM1 antibody blocked CEACAM1-mediated negative regulation and restored T/NK cell activities. Different expression patterns of CEACAM1 compared with PD-L1 and efficacies in a single mouse trial support the rationale for developing anti-CEAEAM1 antibody as cancer therapeutics.
Electron microscopic characteristic of local immune processes during regression of developed experimental tumors under the influence of magnetite nanoparticles.
Authors: Zhukova Galina V. , Kit Oleg I. , Bragina Marina I. , Gudtskova Tatiana N. , Shikhlyarova Alla I. , Shirnina Elena A. , Zhadobina Anastasia I. , Zernov Vladimir Alekseevich , Rudenko Mikhail Yu.
Published: May 2019

e14179

Background: Previously, regression of the developed transplanted tumors under the influence of magnetite nanoparticles (NPs) as monofactor was shown. The purpose of the study was to reveal signs of intercellular interactions in the zone of tumors regressed under the influence of magnetite NPs. Methods: Experiments were carried out on 14 white outbred male rats (180-200 g) with transplanted sarcoma 45 of an initial volume of 0.7-1.3 cm3. After 6 peritumoral injections of magnetite NPs (10 ± 2 nm) in the form of magnetic fluid AM-01 ("AM-Kub", Ekaterinburg) twice a week in a single dose of 17.7 mg/kg, 5 out of 7 rats of the main group demonstrated a regression of the tumor from 1.5-3 cm3 to 0.7 ± 0.2 cm3 (in the control group – 9.7 ± 1.9 cm3, p < 0.01). Ultrathin sections of the tumor tissue of 12 animals (including 7 rats of the main group) were examined in electron microscope JEOL JEM-1011 (Japan). Results: Under effective action of magnetite NPs in tumor tissue, in addition to necrosis, apoptosis and autophagy were marked. In the case of apoptosis, chromatin condensation in compact masses and separation of apoptotic bodies were observed. Significantly more common autophagic cell death was identified by violation of the integrity of the cell membrane and the presence of autophagosomes. Numerous signs of activation of cell-cell interactions with participation of immune cells were observed. Diverse groups of 2-4 contacting cells included macrophages, lymphocytes, plasmocytes, degranulated mast cells, neutrophils in various combinations with distinct signs of the metabolic activity. Invagination of macrophages cytoplasm into the cytoplasm of tumor cells, close adjacency of the cytoplasmic membranes of neutrophils and lymphocytes at considerable length, tight intertwining of mast cell cytoplasmic processes and engulfing of the mast cell granules by macrophages were noted. Conclusions: The results supplement the characteristics of immune processes during the self-dependent antitumor action of magnetite NPs.
Characterizing the gut microbiome of patients with triple-negative breast cancer.
Authors: Swoboda April T. , Sharma Anukriti , Olopade Olufunmilayo I. , Nanda Rita , Gilbert Jack
Published: May 2019

e14186

Background: The human microbiome has been postulated to play a role in carcinogenesis, and there is evidence that commensal intestinal bacteria are capable of promoting tumor immunosurveillance and controlling the immune system’s response to therapy by modulating the tumor microenvironment. Methods: We characterized the stool microbiome of 30 patients (pts) with triple-negative breast cancer (TNBC) using 16S rRNA gene sequencing. Analysis of composition of microbiomes (ANCOM) was used to identify differential abundant exact-sequence-variants (ESVs) with p-values adjusted using Benjamini-Hochberg False Discovery Rate (pFDR) correction. We correlated the stool microbiome with clinicopathologic features and response to treatment. Results: The cohort included 7 pts with metastatic breast cancer and 23 pts with early-stage disease. 19 pts were treated with neoadjuvant chemotherapy (NACT). Bacteroides ovatus ESV was enriched (pFDR < 0.05) in black pts (n = 14), whereas ESV from Prevotella copri was abundant (pFDR < 0.05) in white pts (n = 13). ESVs from Clostridium and family Ruminococcaceae were enriched (pFDR < 0.05) in pts with distant metastatic recurrence (n = 6) compared to pts without metastatic disease. Notably, we identified specific taxa that differentiated between pts with a pathological complete response to NACT (pCR, n = 6) and those with residual disease (n = 13). ESVs from Bacteroides and Ruminococcaceae were relatively more abundant in pts with a pCR (pFDR < 0.05). Furthermore, among pts with residual disease after NACT, Bacteroides caccae was significantly enriched in pts with a partial response (n = 4) compared to pts with no response (n = 9). Conclusions: To our knowledge, this is the first time that 16S rRNA amplicon sequencing has been used to identify microbial signatures correlating with clinical phenotypes and response to treatment in pts with TNBC. This small study provides insight into the potential role of microbes in novel therapeutic interventions to correct dysbiosis and shift towards a gut microbial ecosystem that enhances the body’s immune response to TNBC.
Recapitulation of inflammatory and immune-evasive subtypes of oral cancer cells in immunodeficient mice.
Authors: Lin Su-Fang , Ko Ying-Chieh , Wang Fu-Hui , Su Sheng-Yao , Chen Yu-Lian , Lai Ting-Yu , Liu Yi-Hong , Lin Chung-Yen , Hsu Shu-Ching
Published: May 2019

e14199

Background: Prior study stratified Taiwanese oral cancer specimens (n = 40) and cell lines (n = 7) into three distinct molecular subtypes, namely classical (CL), mesenchymal (MS), and basal (BA). In a cell line derived xenograft (CDX) model, we observed MS grew at least 10-fold slower than CL did in immunodeficient mice. By using RNA-seq to portrait the transcriptomes of human tumor and mouse stroma simultaneously, contribution of mouse innate immunity in restricting the growth of human oral cancer cells was assessed. Methods: A half millions of mycoplasma-free OC3 (MS) or TW2.6 (CL) cells with matrigel were subcutaneously implanted into the flank of 10 to 16 weeks old NOG (NOD/SCID/Il2rgtm). RNAs of CDX tissues from OC3-NOG (n = 7) and TW2.6-NOG (n = 5) were extracted and subjected to stranded mRNA sequencing. Clean reads were aligned to GRCh38 (human) and GRCm38 (mouse), respectively, followed by identifying differentially expressed genes and gene set enrichment analysis. Results: Up-regulation of signature genes for dendritic cells and macrophages and enrichment of innate immunity, including Tnfa-Nfkb signaling, interferon alpha response, interferon gamma response and inflammation were detected in OC3- but not in TW2.6-CDXs. These results suggested that OC3 (MS) was more immunogenic than TW2.6 (CL), which is reminiscent of the inflammatory MS (IMS) subtype of head and neck cancer recently described by Keck et al (n = 938, Clin Cancer Res 2015, 21: p870. PMID: 25492084). Conclusions: By RNA-seq/xenome analysis of CDXs, we provide evidence that compared to CL, MS subtype of oral cancer cells elicited a stronger innate immunity in NOG mouse. Alternatively, CL subtype might have evolved as a prototype with superiority in immune escape.
The role of proinflammatory immune (IFN-gamma) gene signature in predicting prognosis and response to chemotherapy in uterine carcinoma.
Authors: Mahdi Haider , Rose Peter Graham , Abdul-Karim Fadi W , Monk Bradley J. , Ni Ying
Published: May 2019

e14209

Background: Immunotherapy is promising option given low toxicity and potential durable response. In mismatch repair proficient endometrial and ovarian cancers, the reported response rate is ranging from 10-15% in recurrent setting. We need to better identify subset of patients who benefits from immunotherapy. Multigene immune signatures represent a robust means of capturing a complex, T cell–inflamed phenotype necessary for the clinical activity of PD-1–/PD-L1–directed monoclonal antibodies. IFN-γ is a key cytokine produced by activated T cells, as well as natural killer (NK) and NK T cells, in the tumor microenvironment. An immune-related IFN-γ 18-gene profile was derived through a cross-validated penalized regression modeling strategy to predict response to anti-PD1 therapy across 9 different tumor types. We want to test if this gene panel can also predict cancer outcome and response to chemotherapy. Methods: We used whole transcriptome sequencing of RNA matched tumor-normal samples from 38 high stage (Stage III and IV) uterine serous cancer patients. All patients received chemotherapy with platinum and taxanes. IFN-18 gene expression score was calculated by averaging the normalized and log transformed individual gene read counts. The optimized score cut off was selected to best separating the progression free survival. Then the cut off score was tested in The Cancer Genomic Atlas (TCGA) uterine and ovarian cancer RNAseq datasets. Results: The IFN score of 2.46 was determined based on 18-gene expression derived from 38 high-stage uterine serous cancer samples. Average age was 67 years (range: 56-82 years). Uterine serous cancer is known to be MSI stable. Patients with score higher than 2.46 showed significantly longer progression free survival (PFS – 57.6 months vs 15months, p = 0.002) and longer overall survival (73.1 months vs 51.1 months), not statistically significant given our small sample size, p = 0.13) compared to the patients with score lower than 2.46. Then this IFN based gene signature was then applied to TCGA 541 uterine cancer samples with RNAseq data. Similarly, this signature predicted significant improvement in both progression-free survival (p = 0.001) and overall survival (p = 0.005). Interestingly, this score cannot separate outcome for TCGA ovarian cancer cohort. Conclusions: Immune-related IFN-γ gene signature predicted prognosis and response to chemotherapy. We plan to assess if this signature will predict endometrial cancer patients who benefits from anti-PD1 therapy.
Oncolytic and protective effect of strain no. 228 of a new rotavirus group in the Reoviridae family on В16/F10 melanoma growth in experiment.
Authors: Kolpakov Sergey A. , Kolpakova Elena P. , Zlatnik Elena Yu. , Sitkovskaya Anastasia O. , Shulgina Oksana G. , Zolotareva Ekaterina I. , Kopyltsov Vladimir N.
Published: May 2019

e14210

Background: The purpose of the study was to determine the effect of a strain No. 228 of a new rotavirus group of the Reoviridae family on the growth of transplantable В16/F10 melanoma in mice and survival of animals. Methods: The study was performed using a strain No.228 of a new group of rotaviruses (working title «group K», RVK) ( http://jbks.ru/archive/issue-10/article-6 ). RVK is a live attenuated and apathogenic strain growing on the PEKC culture (pig embryo kidney cells) with 5·109 viral particles/mL. The dynamics of melanoma growth and survival of animals receiving RVK were studied on 25 male mice divided equally into 5 groups. RVK was administered intramuscularly and Per os once a week, respectively 0.3 mL and 25 µL to groups 1-2 21 days prior to the tumor inoculation, a total of 4 injections (“vaccination” regimen); to groups 3-4 – 7 days after tumor inoculation with the appearance of tumor nodes, a total of 3 injections (“treatment” regimen); group 5 – controls receiving saline. Groups 1 and 3 received live RVK, groups 2 and 4 - RVK inactivated by UV radiation. Results: Survival in both groups of animals with “vaccination” regimen was improved significantly, compared to controls (group 1 – 25.8±2.2 days after melanoma transplantation; group 2 – 25.4±1.1 days; controls – 14.6±3.08 days, p < 0.05). In groups with “treatment” regimen, survival was 19.2±3.08 days in group 3 and 21.6±3.52 days in group 4, the latter significantly higher than in controls (p < 0.05). Introduction of a live virus in the “vaccination” regimen resulted in significantly better survival compared to the “treatment” regimen (group 1 vs. group 3), while inactivated virus did not caused such a difference (groups 2 and 4). Conclusions: Improved survival of mice with B16/F10 melanoma after administration of RVK demonstrated its ability to inhibit tumor growth and confirmed our previously published data obtained on Guerin's carcinoma in rats. Administration of a live virus prior to the tumor inoculation cause stronger effect than after inoculation. Since inactivation of the virus does not eliminate its effect, apparently, it is associated with immunostimulating action, rather than with oncolytic one.
Recycling of residual liquid drug formation following pressurized intra-peritoneal aerosol chemotherapy (PIPAC).
Authors: Chaudhry Haris , Khosrawipour Tanja , Aslam Saher , Mikolajczyk Agata , Schubert Justyna , Pigazzi Alessio , Khosrawipour Veria
Published: May 2019

e14215

Background: Pressurized Intraperitoneal Aerosol Chemotherapy (PIPAC) is becoming an increasingly widespread approach in which intra-peritoneal chemotherapy (IPC) is delivered in the shape of a chemoaerosol formed by an aerosol creating device. Disadvantages of current PIPAC applications include nearly instant liquid drug accumulation in the recto-uterine pouch. This study aims to investigate the feasibility of respraying the accumulated liquid to prevent local cytotoxicity and tissue trauma as well as to possibly adjust drug concentrations and improve both distribution patterns and penetration levels. Methods: A well-established, previously described postmortem swine model was used. 200ml of doxorubicin solution were aerosolized within the abdominal cavity at a pressure of 12 mm Hg C02 and 37° degrees Celsius. A suction tube was placed in the recto-uterine pouch and accumulating liquid doxorubicin solution following aerosol formation was collected and recycled in consecutive PIPAC repetitions. PIPAC repetitions were performed by using only the remaining liquid drug within the pouch. Consecutive residual volumes within the recto-uterine pouch were measured, as well as tissue penetration rates at different locations of the peritoneal wall. Results: Liquid formation was continuously reduced with each consecutive spray cycle. Tissue penetration rates were significantly higher following the recycling process (p < 0.05). Drug distribution pattern showed a significant improvement following the recycling process vs. after initial aerosol formation (p < 0.05). Conclusions: Respraying the accumulated liquid drug in the recto-uterine pouch is both feasible and important to eliminate liquid accumulation following PIPAC application. Preliminary data gathered in this study suggest that such a recycling process both improves drug distribution and increases penetration levels. However, recycling might also increase the technical complexity of the entire applicational procedure.
A novel dual inhibitor of IDO and TDO, CMG017, potently suppresses the kynurenine pathway and overcomes resistance to immune checkpoint inhibitors.
Authors: Kim Chan , Kim Joo Hoon , Kim Jin Sung , Chon Hong Jae , Kim Joo-Hang
Published: May 2019

e14228

Background: Kynurenine production by indoleamine 2,3-dioxygenase (IDO) is critical for tumor immune suppression through effector T cell anergy and regulatory T cell proliferation. This has led to the rapid development of IDO inhibitors for cancer immunotherapy. However, results from recent clinical trials have been disappointing and this is partly due to pathway redundancy. Tryptophan 2,3-dioxygenase (TDO), another important enzyme of the kynurenine pathway, plays a compensatory role in the absence of IDO activity. Therefore, we developed a dual inhibitor of IDO and TDO to achieve maximal inhibition of the kynurenine pathway and alleviate tumor immune suppression. Methods: Small-molecule inhibitors of IDO and TDO were synthesized and evaluated using in vitro cell-based assays. Pharmacokinetic and pharmacodynamic profiles were assessed for these inhibitors. Tumor-bearing mice were treated with CMG017 per os, either alone or in combination with immune checkpoint inhibitors (ICIs). The tumor microenvironment (TME) was assessed through histological, flow-cytometric, and Nanostring immune profiling analyses. Results: CMG017 suppressed kynurenine production more effectively than inhibitors targeting either IDO or TDO alone, in various human and murine cancer cell lines. Single administration of CMG017 showed favorable pharmacokinetic profiles compared with an IDO1 selective inhibitor. Repeated once-daily per os administration of CMG017 decreased kynurenine concentration in both tumors and plasma of tumor-bearing mice and delayed tumor growth without significant toxicity. CMG017 induced dramatic changes in immune-related genes in TME and enhanced intratumoral infiltration of CD8+ effector T cells. The anti-tumor activity of CMG017 was almost negated when T cells were depleted, indicating the importance of adaptive immunity for the in vivo efficacy of CMG017. Of note, combination immunotherapy of CMG017 with ICIs (anti-PD-1 and anti-CTLA-4) led to durable tumor regression and long-term overall survival. Mice with complete tumor regression were immune to tumor re-challenge, indicating the establishment of immunological memory. Conclusions: CMG017, a dual inhibitor of IDO and TDO, potently suppressed the kynurenine pathway and showed promising anti-cancer efficacy, with favorable pharmacologic profiles.
Monoclonal microbial EDP1503 to induce antitumor responses via gut-mediated activation of both innate and adaptive immunity.
Authors: Gardner Humphrey Athelstan , Kashyap Shubhra , Ponichtera Holly , Sandy Peter , Parameswaran Pooja , Carlson Mark , Sizova Maria , Kravitz Valeria , Andrewes Sam , Troy Erin , Bodmer Mark , Francisco Loise
Published: May 2019

e14241

Background: Systemic immunity is regulated by interactions of commensal bacteria with immune cells in the gut. Enrichment of specific intestinal microbes has been shown to enhance the anti-tumor response to PD-1 blockade in both murine models and cancer patients. Here we report that oral administration of a monoclonal microbial, EDP1503, induces systemic anti-tumor immunity. Methods: The mechanism of action and efficacy of EDP1503 was investigated in isograft tumor models and a variety of ex vivo and in vitro studies in murine and human cells. Results: EDP1503 increases expression of costimulatory molecules on CD11c+ dendritic cells (DCs) within the mesenteric LNs with an accompanying increase in proinflammatory CD103+ DCs within tumor draining lymph nodes. In addition, EDP1503 amplifies both myeloid and lymphocyte responses via production of DC-derived growth factors, M1 polarization of macrophages, and production of the lymphocyte-recruiting chemokines, CXCL9 and CXCL10. Mechanistically, EDP1503 triggers specific pattern recognition receptors and induces proinflammatory responses in antigen presenting cells. Moreover, in vivo, treatment of mice with EDP1503 results in decreased tumor volume and delayed tumor growth. The prominent anti-tumor effects of EDP1503 are further augmented by combination with anti-PD-1 neutralizing Abs. Dissection of the tumor microenvironment reveals increased activated Ki67+ NK cells and CD8+ T cells producing IFNg. Conclusions: Together, these data clearly demonstrate the ability of an orally delivered non-colonizing monoclonal microbe to enhance innate and adaptive anti-tumor immunity and substantiates the rationale for ongoing clinical trials. EDP1503 is currently in Phase 1b/2 studies (NCT03775850; NCT03595683) with enrollment open at multiple sites.
A phase II study of cryoablation (cryo) of an enlarging tumor in patients (pts) with advanced lung cancer or melanoma receiving post-progression immune checkpoint inhibition (ICI).
Authors: Mooradian Meghan , Fintelmann Florian , Fadden Riley , Rubin Krista M. , Lawless Aleigha , Vitali Marlena , Sharova Tatyana , Boland Genevieve Marie , Lawrence Donald P. , Cohen Justine Vanessa , Azzoli Christopher G. , Sullivan Ryan J.
Published: May 2019

e14243

Background: Image-guided percutaneous cryoablation is an established minimally invasive oncologic treatment. Through direct modulation of the tumor, it is theorized that this local therapy may modify the immune microenvironment. We hypothesized that it can augment an anti-tumor response when used concurrently with ICI. Methods: In this phase II single-arm study, pts with advanced lung cancer or melanoma progressing on ICI undergo cryo of an enlarging lesion and ICI is continued for a minimum 2 additional cycles. Computed tomography is performed at 4-6 weeks following cryo to determine tumor response in non-ablated lesions per RECIST1.1, with confirmatory scans at 8-10 weeks. The primary endpoint is safety and feasibility. Secondary endpoints are overall response rate (ORR) and disease control rate (DCR) with DCR defined as the percentage of pts who achieve complete response (CR), partial response (PR), and stable disease (SD). Correlative analysis on pre- and post-cryo biopsy specimen, when evaluable, will be performed. A planned analysis to document preliminary safety and feasibility in the first 11 patients was performed. Results: 11 pts (out of planned 40) have been enrolled. 8 with melanoma, 3 with lung cancer. All pts received prior PD-1/PD-L1 monotherapy with 8 pts treated with pembrolizumab, 2 with nivolumab and 1 with atezolizumab. Treated lesions were in lung (n = 4), bone (n = 3), lymph nodes (n = 2), liver (n = 1) and adrenal gland (n = 1). No immune related adverse events occurred. There was one treatment-related CTCAE grade 3 event (osteomyelitis) but no grade 4/5 events. One pt. experienced grade 3 hyponatremia, unrelated to cryo. In evaluable pts (n = 10), ORR was 10% and DCR was 50% (4 SD, 1 PR). Conclusions: During this scheduled safety analysis, cryo following progression on ICI was feasible and had an acceptable side effect profile. Early efficacy data of this potentially synergistic approach is encouraging but warrants further investigation. Clinical trial information: NCT03290677. [Table: see text]
Dipeptidyl peptidase inhibitor BXCL701 synergizes with immunomodulatory agents in a preclinical model of colorectal cancer (CRC) by altering the function of various components of the immune system.
Authors: Agarwal Veena , MacDougall John , Bhatia Dimple , Gupta Snigdha , Jagga Zeenia , Keshava Nirmal , Seth Subhendu , Banga Hemant
Published: May 2019

e14253

Background: The oral DPP8/9 and FAP inhibitor BXCL701 in combination with a checkpoint inhibitor anti-PD-1 antibody (aPD-1) has demonstrated inhibition of tumor growth as well as up-regulation of immuno-stimulatory cytokines and tumor-infiltrating immune cells in animal models [ASCO 2018]. In the present study, BXCL701 was evaluated for its potential to improve synergistically the survival of animals when combined with anti-CTLA-4 antibody (aCTLA-4), an immune checkpoint blocker, and galunisertib, a transforming growth factor (TGF) β type I receptor inhibitor that counteracts the immunosuppressive influence that TGF β signalling has in the tumor microenvironment. Methods: In vivo studies were conducted utilizing the syngeneic colon cancer model (MC38) in C57BL/6 mice. BXCL701 was administered at the dose of 20 µg qd, p.o, while aPD-1 and aCTLA-4 were administered at the dose of 5 and 10 mpk q2w, i.p. respectively. Finally, galunisertib was administered at 75 mpk bid. Dosing was carried out for 4 weeks and median survival of the animals was calculated. Results: BXCL701 demonstrated a significant improvement in the survival of animals when combined with aCTLA-4. The median survival of animals treated with combination of BXCL701 and aCTLA-4 was significantly increased to 51.5 days in comparison to 40 days observed in BXCL701-treated animals (p < 0.001). Furthermore, in a separate study, addition of BXCL701 to the double combination of aPD-1 and aCTLA-4 resulted in a median survival of 51 days in comparison to 37 days for aPD-1 and aCTLA-4-treated animals (p < 0.05). Similarly, addition of BXCL701 to the combination treatment of aPD-1 and galunisertib increased the median survival to 40.5 days in comparison to 28 days observed in animals treated with galunisertib and aPD-1 combination (p < 0.05). Conclusions: BXCL701, an innate immune activator, synergistically combines with aCTLA-4 and a combination of aPD-1 with galunisertib to improve survival response likely by turning cold tumors to hot tumors. The present study advocates for a clinical evaluation of BXCL701 in combination with these immunomodulators in solid tumors.
Concordance between PD-L1 assays for metastatic renal cell carcinoma (mRCC) and metastatic urothelial carcinoma (mUC).
Authors: Labriola Matthew , Zhu Jason , Cheris Sachica , Liu Xin , Perkinson Kathryn , Su Zuowei , McCall Shannon , Huang Jiaoti , Gupta Rajan T. , Armstrong Andrew J. , George Daniel J. , Zhang Tian
Published: May 2019

e14259

Background: Immune checkpoint inhibitors (ICIs) are standard of care for mRCC and mUC patients (pts). PD-L1 status is gaining importance as a predictive biomarker, particularly for cisplatin-ineligible mUC. PD-L1 positivity is defined differently by PD-L1 assay and tumor type, with limited concordance studies. Given real-world limitations in PD-L1 testing, assay concordance studies are needed to distinguish positive (pos)/negative (neg) results and treatment selection. We compared Dako 28–8 and Ventana SP142 assays in mRCC and Dako 22C3 and Ventana SP263 assays in mUC. Methods: 32 pts with mRCC and 18 pts with mUC who had received ICI therapy at Duke Cancer Institute were identified. FFPE archival tumor samples for pts with mRCC were evaluated with Dako 28–8 and Ventana SP142 PD-L1 immunohistochemistry (IHC) assays. For pts with mUC, FFPE archival tumor samples were evaluated with Dako 22C3 and Ventana SP263 PD-L1 IHC assays. Scoring was validated by two pathologists using the scoring system for each assay. PD-L1 status was subsequently correlated to best RECIST response (objective response rate (ORR) defined as stable disease or better). Results: Tissue was obtained from primary tissue in 72% of mRCC cases and in 61% of mUC cases, with remainder from metastatic biopsies. The majority of mRCC cases (29/32, 91%) were concordant between Dako 28-8 and Ventana SP142 assays (8 cases pos and 21 cases neg), with 3 discordant cases (1 case pos for Dako 28-8 but neg for Ventana SP142 and 2 cases neg for Dako 28-8 but pos for Ventana SP142), all from primary tissue. The majority of mUC cases (17/18, 94%) were also concordant between Dako 22C3 and Ventana SP263 assays (2 pos cases and 15 neg cases), with 1 indeterminate Dako 22C3 test on a metastatic biopsy due to background lymph node. In mRCC, the ORR for PD-L1 pos cases was 45% (5/11) versus 33% (8/24) for PD-L1 neg cases. In mUC, the ORR for PD-L1 positive cases was 50% (1/2) versus 31% (5/16) for PD-L1 neg cases. Conclusions: There was strong concordance between the clinically meaningful PD-L1 assays chosen for comparison in both mRCC and mUC. mUC results were limited by low PD-L1 expression in this cohort. Although PD-L1 status does not fully predict for response to ICIs, this suggests that PD-L1 testing could be used interchangeably for the majority of cases when selecting ICI treatment in mRCC and mUC.
Cancer-testis gene-expression features in various tumors.
Authors: Soldatova Kristina I. , Kit Oleg Ivanovich , Tolmakh Roman E. , Vladimirova Liubov Yu , Kutilin Denis S.
Published: May 2019

e14260

Background: Cancer-testis antigens (CTA) can be an effective target for immunotherapy. Immunotherapeutic approaches targeting CTA in breast cancer (BC), endometrial cancer (EC), ovarian cancer (OC), and colon cancer (CRC) are in the incipient stages of development. The purpose of our study was screening of CTA specific for BC, EC, OC and CRC, based on an analysis of transcriptional profiles of CT-genes. Methods: Tumor and intact tissues of the breast, uterus, ovaries and colon were studied in 35, 30, 20 and 60 patients, respectively. RNAs were isolated using the method described by Chomczynski and Sacchi (2006). The REVERTA-L reagent kit was used for the cDNA synthesis. Relative expression of 16 genes ( MAGEA1, MAGEA2, MAGEA3, MAGEA4, MAGEB1, MAGEB2, GAGE1, GAGE3, GAGE4, MAGEC1, BAGE, XAGE3, NYESO1, SSX2, SCP1, PRAME1) was determined by Real-Time qPCR (with GAPDH and GUSB as reference genes). Statistical analysis was performed using the Mann-Whitney test. Results: BC patients showed significantly (p < 0.005) increased expression of MAGEA3, MAGEA4 and GAGE3 in tumor tissues compared to normal ones; EC patients - significantly (p < 0.05) increased expression of MAGEA1, MAGEA2, MAGEA4, MAGEB2, GAGE3, NY-ESO1, SCP1 and PRAME1 in tumor tissues compared to normal ones; OC patients - significantly (p < 0.05) increased expression of MAGEB1, MAGEB2, GAGE1, NY-ESO1 and decreased expression of MAGEA3, MAGEA4, GAGE3, GAGE4, XAGE3, SSX2, SCP1 and PRAME1; CRC patients - significantly (p < 0.05) increased expression of SSX2 and PRAME1, together with decreased BAGE expression, in tumors compared to normal tissues. Conclusions: Analysis of the transcriptional activity of CT-genes revealed the most common diagnostic markers and immunotherapeutic targets for every malignancy: in BC - MAGEA3, MAGEA4 and GAGE3, in EC - MAGEA1, MAGEA2, MAGEA4, MAGEB2, GAGE3, NY-ESO1, SYCP1 and PRAME1, in OC - MAGEB1, MAGEB2, GAGE1 and NY-ESO, in CRC - SSX2 and PRAME1.
PD-L1 expression in soft tissue sarcomas and its prognostic implication.
Authors: Kelany Mohamed Reda , Barth Thomas , Salem Dina , Mosaad Marwa
Published: May 2019

e14261

Background: The expression rate of PD-L1 in human malignant tumors paid a lot of attention and has been reported to vary from 19% to 92%, and the expression of PD-L1 was associated with poor prognosis of various human and may predicted the response to immune checkpoint inhibitors. The purpose of this study to investigate the expression of PD-L1 and its clinic-pathological feature in soft tissue sarcomas (STS). Methods: We collected Paraffin embedded tissue of 50 Patients with STS retrospectively from the archives of Pathology and oncology Department, Ain Shams University Hospitals from 2011-2017.then we correlated between PD-L1 expression and the clinico-pathological features. For PD-L1 expression we followed the Cologne 6-step proportional scoring system, using the rabbit monoclonal antibody (E1L3N 13684) and Tumors with > /10% membranous stained cells were considered to be positive for expression. The differences among variables were calculated by chi-square test. Results: PD-L1 expression was found in 12 (24%) of cases. Of all sarcoma subtypes , desmoid tumors had the highest prevalence of PD-L1 expression noted in 2/3 (66%) of samples followed by GIST 2/4(50%), then liposarcoma 3/11(27%). In addition, there were 2 rhabdomyosarcoma cases with tumor positivity for PD-L1 expression. We found that STS patients with PD-L1 expression have shorter survival with a median OS 11 vs 19 months (p-value = 0.1) as well as a mean PFS 6 vs 11 months (p-value = 0.1) in PD-L1 positive and negative respectively; this finding considered as clinically significant, but not reached to statistical significant . Conclusions: We report here that PD-L1 expression is as prognostic factor for poorer outcome. This finding may be clinically significant but does not reach the statistically significant levels due to small number of samples included. Its role beyond that of a prognostic biomarker remains to be established in the context of larger clinical trials. Clinical trial information: 31/2017.
Evaluation of combined biomarkers for tumor response to immunotherapy (I/O) in patients with advanced non-small cell lung cancer (NSCLC).
Authors: Ramdani Hayat Oum El Kheir , Falk Markus , Schatz Stefanie , Tiemann Markus , Heukamp Lukas , Schuuring Ed , Groen Harry J.M. , Griesinger Frank
Published: May 2019

e14269

Background: Immune checkpoint inhibitors have revolutionized NSCLC treatment. At present, the only established predictive biomarker for I/O-therapy stratification are PD-L1 expression and MSI status. However, the expression of PD-L1 is limited by heterogeneous expression and even high expressors not always respond to I/O therapy. The aim of the study is to evaluate the value of combinations of positive (Tumor Mutational Burden, PD-L1) and negative (a.o. CD73 expression and inactivating STK11 mutations) predictive markers in patients (pts) with advanced NSCLC on I/O therapy. Methods: A retrospective study was performed on a cohort of 54 pts with advanced NSCLC that have been treated with I/O between 2015 and 2017. Pts were selected by the availability of tumor tissue and based on tumor response evaluated by RECIST v1.1 criteria: only patients with durable tumor response (CR,PR ≥ 6 months) and patients with no tumor response (PD as best response) were analyzed for biomarkers: hybrid capture NGS assay for TMB (New Oncology) including STK11 mutations and IHC tests for PD-L1, CD73 and VISTA. Adjusted Cox regression and ROC analysis will be performed to evaluate the predictive value of the different biomarkers. Results: 43/54 pts received nivolumab, 11/54 pembrolizumab in different therapy lines (from 1st to 6th line). 24 pts were defined as having a durable tumor response (median PFS 20 months, median OS not reached) 30 pts as primary progressors (median PFS 2 months, p > 0.0001), median OS 12 months, p > 0.0001). In 30/54 pts enough material was available for TMB testing. The median TMB-value is 11.42 mutations/Mb. In 13 durable responders median TMB-value was 13.28 mutations/Mb versus 11.00 mutations/Mb in 17 primary non-responders. Additional analyses of PD-L1, CD73, STK11 and VISTA will be presented at the meeting as well as correlative data of the parameters analyzed. Conclusions: Our results suggest that integrating several biomarkers including positive and negative predictive markers may correlate better with responses to I/O than PD-L1 and TMB alone.
Correlation between K-RAS mutant subsets, TP53 mutation, and PD-L1 status in non-small cell lung cancer (NSCLC).
Authors: Abdel Karim Nagla Fawzy , Onuoha Chisom , Ahmad Imran , Eldessouki Ihab , Ma Patrick C. , Feldman Rebecca , Mamdani Hirva , Portnoy David Craig , Spira Alexander I. , Liu Stephen V. , Naqash Abdul Rafeh
Published: May 2019

e14272

Background: K-RAS is a non-targetable mutation with no established predictive value. However, PD-L1 is the only approved predictive marker for immunotherapy in NSCLC. K-RAS mutation like PD-L1 has been associated with smoking. TP53 and K-RAS mutations have been identified in aggressive neoplasms. We hypothesized that K-RAS mutant NSCLC has higher PD-L1 expression suggestive of improved response to immunotherapy. Methods: The Caris database from 2016 - 2018 was queried and patients with NSCLC were identified. ≥1% staining for PD-L1 (22c3 antibody) was considered positive. PD-L1 expression as well as K-RAS and TP53 mutation status were analyzed and correlation between different variables were identified using ANOVA. Results: We identified 8,471 patients with NSCLC, where 66% had TP53 mutation and 26.9% had K-RAS mutation; both mutations were identified in 12% of the patients. In the subset of combined K-RAS and TP53 mutations there were 71.48% having positive PD-L1 expression with female predominance 1.4:1. Of the eight main K-RAS mutation subsets, G12C subset was the most common. Compared to other K-RAS subtypes, G12C was associated with the higher occurrence of PD-L1 positivity (62.68%,p < 0.04; Table). Conclusions: In comparison to other subsets of K-RAS mutation, G12C had the highest occurrence of PD-L1 expression, suggestive of improved response to immunotherapy. However, the subset of patients with combined K-RAS and TP53 mutations had a higher occurrence for the PD-L1 positive status of 71.48%. [Table: see text]
Biomarkers of Chinese advanced cancer patients based on real-world data: Actionable genomic alterations and tumor mutational burden.
Authors: Wang HaiTao , Wang Huina , Guo Rongyun , Li Mingwei , Zhang Yanrui , Lou Feng , Cao Shanbo
Published: May 2019

e14280

Background: Biomarkers would enrich patients(pts) with responsiveness to both targeted treatments and immunotherapy which have become a priority. However, molecular heterogeneous in actionable gene alterations and mutational loads need to be explored. In this study, based on a real world setting, Cancer Gene Panel (CGP） analysis was performed in a series samples of Chinese pts with advanced solid tumors, to delineate the landscape of actionable mutations and tumor mutational burden (TMB). Methods: 176 advanced pts were enrolled encompassing 19 common tumor types. All tissue samples were analyzed using next-generation sequencing with Acornmed panel including 808 genes. Results: The frequencies of genetic alterations among the most common driver genes were comparable to those reported by MSK-IMPACT (2018) ( TP53: 51.7% vs 44.8%; APC: 9.7% vs 9.8%; EGFR: 12.5% vs 7.7%; KRAS: 16.5% vs 13.6%; PIK3CA: 4.6% vs 12.0%; VHL: 4.0% vs 6.7%). 58.3% of pts with refractory solid tumors had at least one actionable mutation. Among those pts, 16.7% of them including CDK4, CDK6 or CDKN2A/B could benefit from palbociclib, and 32.1% of them including mTOR, PIK3CA, PTEN or STK11 could benefit from everolimus, and 61.9% of them including BRCA2, ARID1A, MSH1/2/6 or ATM could benefit from olaparib. Across all the specimens, the top 20% of TMB was 17.23 mutations/Mb, with a range of 0-48 mutations/Mb. The top 20% of TMB for each tumor type was 21.67(Bladder cancer), 17.23(Lung cancer), 16.45(Colorectal cancer), 11.04 (Renal cell cancer), 16.25 (Prostate cancer) , 19.70 (Hepatobiliary cancer), 14.78 (Esophagogastric cancer), 7.80 (Pancreatic cancer), 25.31 (undefined tumor) mutations/Mb respectively. Conclusions: This study indicates that most of pts with advanced refractory solid tumors have at least one actionable mutation, and matched therapies may confer clinical benefit. TMB fluctuated wildly across different cancer types, illustrating the unequal objective response rate of immunotherapy across various cancer type. Furthermore, CGP analysis in advanced pan-cancer pts can provide opportunities for targeted therapies and immunotherapy, especially in those with refractory cancer.
Utilization of novel highly multiplexed immunofluorescence microscopy technology to understand immunological tumor microenvironments in small cell lung carcinoma patients receiving combination PD-L1 and PARP inhibition therapy.
Authors: Trindade Christopher Julius , McDonough Elizabeth , Hanson Jeffery , Walter Rodriguez Beatriz , Roper Nitin , Gasmi Billel , Roque Claire , Gebregziabher Makda , Ylaya Kris , Fetsch Patricia , Abdul Sater Houssein , GINTY FIONA , Hewitt Stephen M. , Thomas Anish
Published: May 2019

e14289

Background: Immune checkpoint inhibitors have changed the way we treat most malignant tumors especially lung carcinomas. While PD-L1 is the only FDA approved companion diagnostic used to predict response, its accuracy and specificity is lacking. Thus, new biomarkers and mechanisms of checkpoint inhibitor need to be elucidated. A major limitation with the current approved method, immunohistochemistry, is that it can typically only measure one parameter per tissue section. Currently new multiplexing microscopy techniques are being developed that allow us to see multiple parameters at the cell level, which can answer new questions regarding tumor immune microenvironments (TIME). In this study we used a novel multiplexed immunofluorescent (MxIF) microscopy technique, Cell DIVE™ (GE Healthcare, Issaquah, WA), to analyze formalin fixed paraffin embedded (FFPE) tissue samples from small cell lung carcinoma (SCLC) patients treated with a PD-L1 inhibitor, durvalumab (MEDI4736), in combination with a PARP inhibitor, olaparib. Methods: Biopsies from patients enrolled in a phase II expansion cohort clinical trial (NCT02484404) treated with durvalumab and olaparib were analyzed. Tumor immune response from a cohort of 3 responders and 3 non-responders, before and after treatment, underwent MxIF analysis using biomarkers including CD3, CD4, CD8, CD20, CD39, CD45RA, CD45RO, CD56, CD57, CD103, FOXP3, GATA3, LAMP1, PD-L1 and T-bet. Our panel was able to effectively discern T-cells (including subsets, maturation, and function), B-cells, and NK cells and their interaction with tumor cells. The results were validated for accuracy and consistency using immunohistochemistry and Vectra Polaris Imaging System. Results: Spatial analysis of TIME showed responders (3/3) were associated with an inflamed CD8+ T-cell tumor microenvironment while non-responders (3/3) displayed an excluded or desert CD8+ T-cell tumor microenvironment. MxIF/Cell DIVE™ allowed us to further characterize the immune response. Conclusions: Analyzing the type and pattern of immune response using multiplexed immunofluorescent and single cell characterization microscopy may better predict prognosis than PD-L1 immunohistochemistry in SCLC patients treated with immune checkpoint inhibitors. Clinical trial information: NCT02484404.