bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic differences in tumor-associated macrophages between metastatic and primary sites of clear cell renal cell carcinoma.
Authors: Miura Yuji , Motoshima Takanobu , Wakigami Nanako , Kusada Natsuki , Okaneya Toshikazu , Inoshita Naoko , Takano Toshimi , Kamba Tomomi , Komohara Yoshihiro
Published: February 2018

105

Background: Tumor-associated macrophages (TAMs) are one of the key contributors to the tumor microenvironment and are phenotypically differentiated into M1 and M2 macrophages. M1 macrophages stimulate anti-tumor immune responses, whereas M2 macrophages promote immunosuppression and are associated with tumor progression in clear cell renal cell carcinoma (ccRCC). However, little information is available regarding the difference in TAM polarization between primary and metastatic lesions of ccRCC. Methods: We collected paired samples of primary and matched metastatic sites from the first recurrence in 41 metastatic ccRCC patients. Immunohistochemistry (IHC) for Iba1, which is a common pan-macrophage marker, and CD163 and CD204, which are considered to be M2 macrophage markers, was performed on all paired samples. Results: Thirty-three paired primary and metastatic samples were available for IHC assessment in this analysis. The most common metastatic sites were lung (N = 26, 78.8 %) and lymph node (N = 3, 9.1 %). The mean (± standard deviation) cell density of Iba1+ TAMs was higher in metastatic lesions than in primary lesions (756 ± 267 mm2 vs. 581 ± 155 mm2, P = 0.0012). By contrast, the ratio of CD163+ and CD204+ to Iba1+ TAMs was significantly lower in metastatic lesions than in primary lesions (0.76 ± 0.30 vs. 0.90 ± 0.24, P = 0.0067 and 0.39 ± 0.27 vs. 0.67 ± 0.29, P = 0.0001, respectively). The median overall survival of patients with high- vs. low-density Iba1+, CD163+, and CD204+ TAMs in metastatic lesions was 120 vs. 92 months (log-rank P = 0.67), 120 vs. 58 months (log-rank P = 0.056), and 120 vs. 92 months (log-rank P = 0.35), respectively. Conclusions: TAMs in the metastatic lesions of ccRCC polarized towards an M1-like phenotype, although the total number of TAMs was greater in metastatic compared with primary lesions. The cell density of Iba1+, CD163+, and CD204+ TAMs in metastatic sites was not associated with overall survival in patients with ccRCC.
Serial changes in PD1/PDL1 expression in metastatic urothelial carcinoma (mUC) patients (pts) treated with immune checkpoint blockade (CPB).
Authors: Tzeng Alice , Diaz-Montero C. Marcela , Rayman Patricia A. , Kim Jin Sub , Pavicic Paul G , Finke James , Barata Pedro C. , Lamenza Marcelo , Devonshire Sarah , Schach Kim , Emamekhoo Hamid , Ernstoff Marc S. , Hoimes Christopher J. , Rini Brian I. , Garcia Jorge A. , Gilligan Timothy D. , Ornstein Moshe Chaim , Grivas Petros
Published: February 2018

109

Background: Since CPB may alter immune marker expression in key immunomodulatory populations such as myeloid-derived suppressor cells (MDSC) and CD8+ cytotoxic T lymphocytes (CTL), we evaluated PD1/PDL1 expression in longitudinal samples from mUC pts treated with CPB. Methods: Serial peripheral blood samples were collected from mUC pts who received CPB. PD1/PDL1 and VISTA expression was measured in MDSC (CD33+HLADR−) and CTL (CD8+CD4−) from live peripheral blood mononuclear cells using flow cytometry. MDSC subsets were further defined as (G)ranulocytic (CD15+CD14−), (M)onocytic (CD15−CD14+), and (I)mmature (CD15−CD14−). PD1/PDL1 and VISTA expression was presented as % of each MDSC subset or CTL. Wilcoxon signed-rank tests and mixed-model regression analyses were performed to assess changes in immune marker expression after CPB. Results: Of 30 CPB-treated pts with ≥ 2 blood samples for analysis, 21 received anti-PDL1 (20 atezolizumab/1 avelumab; [A]) and 9 received anti-PD1 (pembrolizumab [P]). Median age at diagnosis was 69.5 (4681), 77% men, 33% never smokers, 63% pure UC, 70% bladder primary, 20% prior intravesical BCG, 37% prior neoadjuvant chemotherapy, 63% prior cystectomy. Best overall responses to CPB were 3 PR/13 SD/5 PD (A) and 1 CR/1 PR/4 SD/3 PD (P). Successive doses of A correlated with decreased %PDL1+ M-MDSC, while those of P correlated with decreased %PD1+ M- and I- MDSC (Table). No significant changes in VISTA expression were detected. In 11 A-treated pts with samples before/after the 1st dose, %PDL1+ M- and I- MDSC decreased (median change −25.5 and −5.7; p = 0.02 and 0.03) and %PD1+ CTL increased (median change +2.4; p = 0.02) between 1st and 2nd samples. Conclusions: In this mUC pt cohort, distinct post-tx changes in %PD1/PDL1 in MDSC subsets and CTL occurred based on CPB (anti-PD1 vs anti-PDL1). Further analysis of correlations between CPB, immune marker expression, clinicopathologic factors, and outcomes is ongoing in a larger cohort. Mean absolute change in marker expression per dose in pts treated with CPB. [Table: see text]
RNAScope as a platform to detect IDO1 expression in tumor tissue sections.
Authors: Pratta Michael
Published: February 2018

116

RNAScope is a sensitive, specific platform to detect IDO1 expression in tumor tissue sections. M Pratta, M Rupar, P Waeltz, T Burn, G Hollis, M Covington, M Smith, and R Newton. Incyte Corp. Wilm. DE. Background: Indoleamine 2,3-Dioxygenase 1 (IDO1) catalyzes the primary and rate-limiting step in tryptophan catabolism to generate N-formyl-kynurenine (Kyn). Through a combination of local depletion of tryptophan and an increase in Kyn concentrations, IDO-1 activity can result in the suppression of antitumor immune responses. Because IDO-1 inhibitors are now in the clinic for treatment of multiple tumor types, immunohistological approaches are employed to demonstrate IDO1 expression in tumor biopsies. However, using a commercially available antibody to detect IDO1 by immunohistochemistry (IHC), the level of sensitivity was inadequate. Methods: In order to improve the sensitivity of IDO1 detection, we evaluated in situ hybridization (ISH) using RNAScope technology and digital quantitation by HALO analysis in collaboration with Advanced Cell Diagnostics (ACD). The technology was cross-validated using IDO1 qRT-PCR, Western blot, and activity analysis and compared with standard IHC. We initially evaluated IDO1 expression in HeLa cells stimulated with various concentrations of IFNγ, and then extended the observations using tissue sections from multiple tumor types. Results: In the HeLa cell model, IFNγ induced a time- and concentration-dependent increase of IDO1 at the mRNA, protein, and activity level. Although IDO1 was successfully detected in the HeLa cell samples by IHC, comparison of the platforms indicated IFNγ EC50 values were in strong agreement between RNAScope (193.8 pg/ml) and Western blot analysis (170.8 pg/ml), but was much higher by IHC analysis (2206 pg/ml). A strong positive correlation (*p < 0.0001) between RNAScope and Western blot analysis was observed, suggesting a highly coordinated induction of IDO1 by IFNγ at both the mRNA and protein levels. FFPE tumor tissue from melanoma, HNSCC, bladder, renal, ovarian, and lung cancers visualized by RNAScope all show varying levels of IDO1 expression. Conclusions: These data support the use of RNAScope for the analysis of IDO1 expression in clinical trials.
Clinical significance of PD-L1 levels in plasma-derived exosomes in head and neck cancer.
Authors: Theodoraki Marie-Nicole , Yerneni Sai Gopal Krishna , Whiteside Theresa
Published: February 2018

117

Background: Despite the broad spectrum of available therapies, HNSCC patients have poor outcome due to locoregional recurrence. Mechanisms responsible for immune suppression include activities of checkpoint receptors/ligands. HNSCCs with high expression levels of PD-L1 have especially poor outcome. Thus, PD-1 and PD-L1 inhibitors show potential as anticancer strategies. However, many patients with PD-L1+ HNSCC don’t benefit from checkpoint inhibitor therapy, suggesting that PDL-1 expression on tumors is not the only factor predictive of response. Tumor derived exosomes carry numerous immunosuppressive molecules and deliver them to recipient immune cells. We have demonstrated that elevated levels of circulating immunosuppressive exosomes play a key role in immune suppression and disease progression. Here, we show that surface PD-L1 on exosomes is responsible for these effects. Methods: Exosomes were isolated from plasma of 40 HNSCC patients by mini size exclusion chromatography, captured on beads using anti-CD63 Abs, stained for PD-1 and PD-L1 and analyzed by flow cytometry. Percentages of PD-L1+ and PD-1+ exosome/bead complexes were correlated with clinicopathological data. PD-L1high or PD-L1low exosomes were incubated with activated human CD8 T-cells ± PD-1 inhibitor and CD69 expression levels on T-cells were measured. Patients’ plasma was tested for soluble PD-L1 by ELISA. Results: The PD-L1 surface expression on exosomes correlated with patient’s disease activity, UICC stage and lymph node status with p values ranging from 0.0008-0.013. In contrast, plasma PD-L1 levels didn't correlate with any parameters. Exosome PD-1 levels were not informative. T-cell activation was inhibited (p < 0.03) by co-incubation with PD-L1high but not PD-L1low exosomes. This inhibition was reversed by adding a PD-1 inhibitor to T-cells prior to their incubation with exosomes. Conclusions: We show that PD-L1 levels on exosomes, but not plasma levels, correlated with clinicopathological data in HNSCC patients. Blocking of PD-L1+ exosomes signaling to PD-1+ T-cells with anti-PD-1 Ab attenuated immune suppression. Altogether, PD-L1+ exosomes serve as useful metrics of disease and immune activity in HNSCC patients.
Interleukin-12 armored chimeric antigen receptor (CAR) T cells for heterogeneous antigen-expressing ovarian cancer.
Authors: Yeku Oladapo O. , Purdon Terence , Spriggs David R. , Brentjens Renier J.
Published: February 2018

12

Background: Immune escape via downregulation of tumor associated antigens (TAAs) is an important mechanism of resistance to Chimeric Antigen Receptor (CAR) T cell therapy. Particularly in solid tumor malignancies where antigen expression could be heterogeneous, the risk of antigen-low or antigen-negative relapse is significantly high. One strategy to overcome this limitation is to reengineer CAR T cells to engage other arms of the immune system such as endogenous cytotoxic T cells and dendritic cells (DC) to broaden the antitumor response beyond the TAA targeted by CAR T cells. This could be achieved by co-modifying CAR T cells with Interleukin-12 (IL-12). IL-12 is a proinflammatory cytokine produced by DCs, and macrophages, and has been shown to promote maturation of DCs and increase T-cell proliferation. We hypothesized that CAR T cells genetically engineered to constitutively secrete IL-12 will be efficacious against Muc16ecto low (MLo) and Muc16ecto high (MHi) heterogeneous tumors in a syngeneic mouse model of ovarian peritoneal carcinomatosis. Methods: ID8 mouse ovarian cancer cells with either low endogenous Muc16ecto or transduced to express high levels of Muc16ecto were generated. Mouse T cells were transduced with plasmids encoding second generation Muc16 or Muc16/IL-12-directed CARs. C57BL/6 mice were inoculated i.p with tumor cells and subsequently treated with CAR T cells. Results: Second generation and IL-12 armored CAR T cells (4H1128?-IL12) were cytotoxic against both MLo and MHi cells in vitro. However, 4H1128?-IL12 were significantly more efficacious at killing both MLo and MHi cancer cells. In vivo, treatment with 4H1128?-IL12 led to significantly improved survival in mice inoculated with a 50:50 mix of MLo and MHi cells. Peritoneal washes performed on mice that succumbed to disease showed equivalent eradication of MLo and MHi. Treatment with 4H1128?-IL12 resulted in increased mature peritoneal DC’s (CD11b+ MHCII+). Finally, surviving mice from 4H1128?-IL12 cohorts were found to have increased T-cell receptor (TCR-β) productive clonality. Conclusions: IL-12-secreting CAR T cells are efficacious against tumors with low and heterogeneous antigen expression.
Patient-reported outcomes in PD-1/PD-L1 inhibitor registration trials: FDA analysis of data submitted and future directions.
Authors: Howie Lynn Jackson , Singh Harpreet , King-Kallimanis Bellinda , Roydhouse Jessica , Theoret Marc Robert , Blumenthal Gideon Michael , Kluetz Paul Gustav
Published: February 2018

134

Background: Patient-reported outcome measures (PROs) can capture the patient’s experience with disease and treatment. Anti-PD-1/PD-L1 therapies have unique symptomatic side effects; PRO data can help to better understand the patient experience on therapy. Health-related quality of life (HRQL) components most impacted by therapy include disease symptoms, symptomatic toxicity and physical function. Methods: We reviewed FDA registration trials for 5 immunotherapy agents (anti-PD-1/PD-L1) to evaluate trial design and PRO assessment. We assessed whether the PRO strategy assessed physical function and symptomatic immune-related adverse events (irAEs) by reviewing whether trials used a well-defined physical function domain and 8 symptoms related to irAEs reported in product labels (fatigue, diarrhea, cough, shortness of breath, musculoskeletal pain, rash, pruritis and fever). Results: Data from 25 trials across 7 disease types and 1 tumor agnostic indication were evaluated. Of these, 13 were randomized and 22 were open label. Eighteen of 25 contained PRO assessments and all 18 used > 1 instrument. The most common instruments were the EQ-5D (N = 17), followed by EORTC QLQ-C30 (N = 15). Disease-specific PRO tools were included in 8 trials (5 lung, 1 head and neck, 1 melanoma and 1 renal cell), consisting of modules or scales from EORTC (N = 5), FACIT (N = 2) or the Lung Cancer Symptom Scale (N = 1). Sixty percent of the trials (15/25) used an instrument that contained a well-defined physical function (PF) domain. No trial used a PRO strategy assessing all 8 selected symptoms related to irAEs. Conclusions: Collection of PRO data in anti-PD-1/PD-L1 trials submitted to FDA was variable, and did not consistently assess treatment related symptoms and physical function. Use of a HRQL tool with well-defined functional scales supplemented by item banks or libraries to incorporate symptoms associated with irAEs may improve understanding of the patient experience while receiving anti-PD-1/PD-L1 treatment. These data, along with other important clinical data such as hospitalizations, ER visits and supportive care medications can inform the benefit risk assessment for regulatory purposes.
Safety and efficacy of checkpoint inhibition (CI) in cancer patients (pts) with concurrent human immunodeficiency virus (HIV) infection.
Authors: Park Soo , Sacco Assuntina Gesualda , Cohen Ezra E.W. , Daniels Gregory A.
Published: February 2018

136

Background: There is limited knowledge about the use of CI in cancer pts with HIV. Here we report the safety and efficacy of these agents in this pt population. Methods: We identified cancer pts with HIV at UCSD who received ≥1 dose of ipilimumab, pembrolizumab or nivolumab (anti-PD1), or the combination of ipi+nivo and had ≥1 follow-up visit. Data regarding clinical history, treatment response, and toxicity were collected. Results: Three pts had head and neck squamous cell carcinoma (HNSCC), 2 pts had advanced melanoma, 1 pt had cutaneous SCC, and 1 pt had SCC of unknown primary (suspected lung). Age ranged from 51-73 years. All pts had ECOG PS 0-1 at time of treatment. Six pts received anti-PD1 for a median of 8 cycles (range 2-35); one pt received 4 cycles of ipi+nivo q3wk followed by 4 cycles of anti-PD1. All 7 pts were on anti-retroviral therapy (ART) and had well-controlled HIV (viral load < 400 copies/mL) with varying degrees of immune dysfunction: 4 pts with moderate immune dysfunction (100-199), 1 pt with mild immune dysfunction (200-350), and 2 pts with normal immune function ( > 350). 3/7 pts (43%) had ≥1 clinically significant irAE of grade ≤2 (fatigue and rash); one pt who received ipi+nivo developed grade 3 hepatitis and required systemic steroids though progressive disease likely contributed. No atypical toxicities were seen. Objective responses were seen in 4/7 pts (57%) with 1 complete response and 3 partial responses; two pts had progressive disease and response was not yet evaluable in 1 pt. Four patients died from clinical deterioration unrelated to CI or immunodeficiency. Conclusions: CI demonstrated significant anti-tumor activity in cancer pts with well-controlled HIV. The toxicity profile is consistent with known irAEs. Further study of these agents in this pt population is warranted.
Immune checkpoint inhibition in elderly non-small cell lung cancer patients.
Authors: Marrone Kristen , Zhang Jiajia , Feliciano Josephine Louella , Forde Patrick M. , Hann Christine L. , Kelly Ronan Joseph , Ettinger David S. , Turner Michelle , Rowe Valerie , Bonerigo Sarah , Smith Kellie Nicole , Anagnostou Valsamo , Velculescu Victor E. , Brahmer Julie R. , Naidoo Jarushka
Published: February 2018

137

Background: Age-related changes in the immune system are dynamic and complex. Use of anti-PD-1/PD-L1 therapy in advanced non-small cell lung cancer (NSCLC), with median patient age of 71, has rapidly become standard of care. Understanding how senescent remodeling of the immune system may affect outcomes with anti-PD-1/PD-L1 therapy is poorly understood. Although trends in phase III clinical trials of anti-PD-1/PD-L1 therapy suggest a potential lack of clinical efficacy in patients ≥ 75 years of age, the proportion of treated patients in this age range was low. The use and outcomes of these agents in this population treated in clinical practice has not been reported. Methods: The Johns Hopkins Upper Aerodigestive Diseases Malignancies and Immunotherapy Database was queried for all patients treated with anti-PD-1/PD-L1 agents as part of a clinical trial or as standard of care, from 2007 to 2017. Patients ≥ 75 years old were assessed for demographic, response, toxicity and survival data as of September 2017. Results: Of the 275 patients in our database, 144 NSCLC patients had received an anti-PD-1/PD-L1 agent alone or in combination with another immune checkpoint inhibitor (52%). Of those 144 patients, 20 patients were ≥ 75 years of age at time of anti-PD-1/PD-L1 therapy initiation (14%), with a median age of 81 years (range: 75-90). Of those 20 patients, 19 received anti-PD1 monotherapy (95%); 1 patient received combination anti-PD-1 with an anti-KIR. 14 of the 20 patients (70%) received therapy in second line or beyond; 6 received anti-PD-1 monotherapy as first line therapy. No significant difference in median survival for 1st line or ≥2nd line group was seen (12.7m vs 15.3m; p = 0.92). A median of 5.5 doses were administered (range 1-24). Reasons for off therapy included: progressive NSCLC (9; 45%), consent withdrawal (1; 5%) and adverse events (6; 30%) with 4 patients who remain on therapy (20%). Conclusions: Anti-PD-1/PD-L1 therapy is now standard of care in NSCLC, a malignancy of the elderly. Translational evaluation of how immune system senescent remodeling affects outcome and toxicity is needed. Further analysis of immunologic and genomic data will be presented and compared to the overall population in our database at the time of presentation.
Whole body PD-L1 PET in patients with NSCLC and melanoma.
Authors: Leung David K. , De Langen Joop , Raunig David , Niemeijer Anna-Larissa N. , Smit Egbert F. , Boellaard Ronald , Vallez-Garcia David , Dongen G.a.M.S. van , Windhorst A.D. , Huisman Marc C. , Glaudemans Andor W. J. M. , Hendrikse N. Harry , Smith Ralph Adam , Poot Alex J. , Lipovsek Dasa , Donnelly David J. , Bonacorsi Samuel J. , Velasquez Linda M. , Du Shuyan , Hayes Wendy
Published: February 2018

139

Background: PD-(L)1 immunotherapy is effective in multiple tumors, including NSCLC and melanoma, but tumor PD-L1 IHC correlates only moderately with treatment outcome. This study aims to assess 1) safety of 18F-BMS-986192 (18F-PD-L1) in human, 2) PD-L1 quantification in tumors using 18F-PD-L1 PET, 3) PD-L1 PET correlation with IHC and treatment outcome, and 4) intra and inter subject tracer uptake variability. Methods: Pts with NSCLC (N = 10) and melanoma (N = 3) were included. At baseline, pts received a static or multiphase dynamic whole body PET scan after injecting 200 MBq 18F-BMS-986192. For NSCLC pts, (1) SUV(max, peak and mean) were measured for each delineable tumor (N = 32, 1-7 tumors/pt), (2) PD-L1 IHC (28.8 assay) was performed on the biopsy, and (3) response to Nivolumab therapy assessed by RECIST 1.1. Intra and inter subject variability and intraclass correlation were calculated using SUVs of all assessed tumors. Equal variance for PD-L1 status was evaluated by a Levene’s test. Four (3 female) pts underwent dosimetry study (ICRP 60). Results: No AEs related to radiotracer was observed. Dosimetry study demonstrated whole body exposure of 30 mGy at dose > 1400 MBq. Biodistribution among pts is comparable. PD-L1 IHC from 13 biopsied lesions were evaluated, 5 < 1%, 4 ≥1%, and 4 ≥50%. Tumor tracer uptake was measured in NSCLC pts and categorized by PDL-1 IHC as ≥50% or < 50%. Clinical trial information: 2015-004760-11. Tumor SUVs did not correlate with RECIST 1.1 assessment. Lesion heterogeneity was reflected in both inter and intra pt variability (CVinter = 41%, CVintra = 53%, ICC = 0.41 for SUVpeak). Levene’s test showed no significance in variability between the two PD-L1 categories. Conclusions: PET-imaging with 18F-BMS-986192 is safe and feasible in pts with NSCLC and melanoma. Pts with higher PD-L1 PET SUV have higher PD-L1 by IHC. Intra pt variability is similar to inter pt variability. With limited number of pts, no clear correlation of PET PD-L1 and tumor response is observed. A prospective study with this tracer is underway to further investigate 18F-BMS-986192 in understanding of PD-L1 expression.[Table: see text]
A single-arm, phase II study assessing the efficacy of pembrolizumab (pembro) plus radiotherapy (RT) in metastatic triple negative breast cancer (mTNBC).
Authors: McArthur Heather L. , Barker Christopher Andrew , Gucalp Ayca , Lebron-Zapata Lizza , Wen Yong Hannah , Phung Anh , Rodine Micaela , Arnold Brittany , Zhang Zhigang , Ho Alice
Published: February 2018

14

Background: Overall response rates of 13-19% have been reported with checkpoint inhibitor monotherapy in chemotherapy-resistant, PD-L1-positive mTNBC. RT is frequently used to enhance local control in mTNBC and has been reported to induce distant (abscopal) tumor responses when combined with immunotherapy. In this study, we evaluate the safety and efficacy of RT combined with a programmed cell death protein 1 (PD-1) inhibitor, pembro, in a single-arm, two-stage, phase II study in mTNBC. Methods: Eligible women had biopsy-proven mTNBC, ECOG performance status 0-2, and ≥2 measurable sites of metastatic disease with at least one site requiring RT. A total RT dose of 3000 cGy was delivered in 5 daily fractions. Pembro 200 mg was given intravenously within 3 days of first RT fraction, then every 3 weeks +/-3 days until disease progression. The primary endpoint was overall response rate at week 13 in the non-irradiated lesions by RECIST v1.1. Secondary endpoints included safety and overall survival. Tumor biopsies were obtained at baseline and at week 7. PD-L1 expression was not required for study entry. Results: Of the 17 women enrolled, the median age was 52 y (range 37-73y). and the median number of prior chemotherapies received for metastatic disease was 3 (range 0 to 8). Of the 8 women not evaluable at 13 weeks: 5 died secondary to disease-related complications (at weeks 2, 6, 7, 8, and 9) and 3 came off study due to disease progression prior to week 13. Of the 9 women evaluable at week 13, 3 (33%) had a partial response, 1 (11%) had stable disease and 5 (56%) had disease progression. The 3 partial responses represented 60%, 54%, and 34% decreases in tumor burden by RECIST v1.1 and were durable for 31, 21, and ongoing at 22 weeks, respectively. The stable disease response was durable for 22 weeks. Common toxicities were mild and included fatigue, myalgia and nausea. Conclusions: The combination of pembro and RT is well-tolerated. This is a poor prognosis population with 5/17 (29%) of patients dying within 12 weeks of study entry. However, durable responses were observed outside of the RT field in 3/9 (33%) patients who were unselected for PD-L1 expression and evaluable at 13 weeks. Clinical trial information: NCT02730130.
Baseline circulating myeloid-derived suppressor cells and response to PD-1 inhibitor in non-small cell lung cancer patients.
Authors: Delaunay Myriam , Guibert Nicolas , Lusque Amellie , Farella Magali , Boubekeur Nadia , Gouin Sandrine , Dormoy Inge , Fons Pierre , Esquerre Michael , Favre Gilles , Pradines Anne , Mazieres Julien
Published: February 2018

145

Background: Inhibitors of immune checkpoint PD-1/PD-L1 (ICI) have become a care standard in non-small cell lung cancer (NSCLC). Despite promising results, some patients cannot take advantage of immunotherapy effects. Nowadays, neither predictive nor prognostic circulating biomarkers have been found in order to select patients or to predict response to ICI. Myeloid-derived suppressor cells (MDSC) are potent immunity suppressors and may represent both a potential prognostic and a predictive biomarker. Methods: Peripheral blood samples regarding 54NSCLC patients treated with nivolumab and 5 healthy donors were prospectively included. Early-MDSC: e-MDSC (LIN-/CD14-/CD15-/HLA-DR-/CD33+), monocytic-MDSC: M-MDSC (HLA-DR low/-/CD14+/CD15-/CD11b+), polymorphonuclear-MDSC: PMN-MDSC (HLA-DR low/-/CD14-/CD15+/CD11b+) were analyzed through flow cytometry. MDSC percentage was calculated from viable peripheral blood mononuclear cells. We assessed if e-MDSC,M-MDSC and PMN-MDSC levels detected before anti-PD-1 therapy correlate with both progression-free survival (PFS) and overall survival (OS). Results: 54 patients were prospectively included: 41 with adenocarcinoma and 13 with squamous NSCLC. When compared to healthy donors, patients had significantly higher levels of circulating e-MDSC(median (range): 0.13% (0-10.9) vs0.02%(0.01-0.14), p = 0.04) and M-MDSC (median (range): 4.55% (0.08-24.18) vs1.36% (0.77-3.76), p = 0.01). Cox proportional hazard regression analysis showed that high level of M-MDSC before anti-PD-1 therapy was associated with OS (HR [CI95%]: 1.08 [1.01; 1.15],p = 0.02) and PFS (HR [CI95%]: 1.07 [1.01;1.14], p = 0.02). No prognostic impact was observed regarding e-MDSC and PMN-MDSC. Conclusions: Our study suggests that a baseline circulating high level of M-MDSC is associated with poor survival. M-MDSC may be a predictive and prognostic biomarker in NSCLC patients treated with anti-PD1. Further studies have been scheduled, including MDSC monitoring during anti-PD1 treatment, in order to confirm our findings. These data would be interesting for the development of combined therapy, targeting the suppressive environment, with ICI.
Correlation of lung cancer mutational profile with immune profile.
Authors: Papanicolau-Sengos Antonios , Pabla Sarabjot , Dy Grace K. , Ernstoff Marc S. , Puzanov Igor , Conroy Jeffrey M. , Nesline Mary , Glenn Sean T. , Burgher Blake , Andreas Jonathan , Giamo Vincent , Qin Maochun , Lenzo Felicia L. , Gardner Mark , Morrison Carl ,   OmniSeq, Inc.
Published: February 2018

146

Background: The association between neoplasm mutational and immune profiles has not been well-characterized. Methods: We collected 26 lung cancer formalin-fixed paraffin embedded (FFPE) samples which had been tested with a comprehensive mutation profile to detect clinically actionable mutations, and a comprehensive immune gene expression profile, which interrogates PD-L1 immunohistochemistry (IHC), PD-L1/2 copy number, CD3/CD8 IHC, microsatellite instability status, mutational burden, and the expression profile of 54 immune-related genes. The ranking of gene expression and 7 immune phenotypes was compared to a reference population. Six cases were positive for an activating KRAS mutation and 2 cases were positive for an activating EGFR mutation. Principal component analysis was performed to determine the association of EGFR/KRAS mutations with the measured immune landscape. Results: The proinflammatory immune phenotype was significantly correlated with KRAS mutation positive samples (first principal component, R squared = 0.53, p < 0.05). Similarly, CD38 expression was correlated with KRAS mutation (R squared = 0.47, p < 0.05). CD137, KLRD1, and DDX58 expression was significantly correlated with EGFR positive samples (second principal component, R squared = 0.47, 0.37, 0.35 respectively, p < 0.05 in all cases). Unsupervised hierarchical clustering of the samples resulted in three distinct clusters, EGFR positive, KRAS positive, and EGFR negative/KRAS negative. In the KRAS positive cluster, high proinflammatory immune phenotype, VISTA moderate expression, presence of CD3/CD8 tumor infiltrating lymphocytes (TILs), and low KLRD1 were overrepresented (p < 0.05), while low VISTA and proinflammatory moderate immune phenotype were significantly underrepresented (p < 0.05). In the EGFR positive cluster DDX58 high, very low TILs, and very low CD8 were significantly (p < 0.05) overrepresented. Conclusions: KRAS mutation positivity is significantly associated with a proinflammatrory immune phenotype and CD3/CD8 infiltration. KRAS positive, EGFR positive, and KRAS/EGFR negative clusters are immunophenotypically distinct. A higher number of specimens is necessary to verify and expand these findings.
Predictive biomarker of response to anti-PD-1 treatment in non-small cell lung cancer patients.
Authors: Zheng Xuan , Hu Yi , Ma Junxun , Zhang Fan , Sun Danyang , Li Lingling
Published: February 2018

148

Background: PD-1 inhibitors have shown significant clinical activity in different cancer types. However, responses in pts with NSCLC are variable, and insights are needed to identify a predictive biomarker of response with greater diagnostic accuracy. Here we tested the hypothesis tha tserum TNF-a level is predictive of response to anti-PD-1 treatment. Methods: NSCLC patients treated with nivolumab or pembrolizumab were studied. Pts received nivolumab (3mg/kg, q2w) or pembrolizumab ( 2mg/kg, q3w). Pts on anti-PD1 were classified as either responders (R) deriving clinical benefit (with SD, PR, CR) or non-responders (NR) not deriving clinical benefit (PD) based on RECIST criteria.Serum was collected at baseline; at 2-3 weeks after the first dose (early stage); and at the time of response evaluation. Serum TNF-a levels were determined by Luminex kit. Changes in serum TNF-a levels and their strength of association with response were compared with Non-parametric Analysis. Results: Evaluable plasma samples were collected from twenty-one NSCLC patients treated with nivolumab or pembrolizumab. There was no significant difference in baseline serum TNF-a levels in responders (n = 15) vs non-responders (n = 6). Between baseline and early stage ,serum TNF-a levels significantly increased in responders (P = 0.010), while in non-responders, no significant change was found. High early change rate of serum TNF-a levels ( > 50%) was observed only in responders(n = 7).At early stage, responders had significantly higher serum TNF-a levels than non-responders(P = 0.008). We found no significant difference in serum TNF-a levels at the time of response evaluation. Conclusions: Early changes in serum TNF-a levels and high serum TNF-a levels at early stage in non-small cell lung cancer patients correlate to response to anti-PD-1 treatment.
The prognostic significance of PDL1 expression in locally advanced non-small-cell lung cancer (LA-NSCLC) treated with definitive intent.
Authors: Mitra Devarati , Agarwal Vishesh , Chen Yu-Hui , Baldini Elizabeth H. , Barbie David Allen , Chen Aileen B. , Glass Benjamin , Herman Gretchen , Kozono David E. , Schoenfeld Jonathan Daniel , Mak Raymond H.
Published: February 2018

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Background: The PACIFIC trial showed an 11-month (mo) improved median (med) progression-free survival after definitive chemoradiation in LA-NSCLC using the PDL1 inhibitor, durvalumab. This study will likely change practice and puts further emphasis on understanding the significance of immune marker expression in this poor-prognosis disease. Methods: From 1999-2012, pathology specimens from 126 patients (pts) with LA-NSCLC who received combined modality therapy at our institution and had sufficient tissue available were selected for tissue microarrays. Immunohistochemistry for PDL1 (E1L3N), PDL2 (9E5), PD1 (NAT105) and CD3 (F7.2.38) was performed and tumor cells and infiltrating immune cells with moderate or high staining intensity were quantified using Definiens Tissue Studio image analysis software. Univariate and multivariate analyses (MVA) were performed using a Cox proportional hazards model. Results: Distribution by stage was: IIA-3%, IIB-3%, IIIA-65%, IIIB-29%. Sixty-three percent of pts had surgery. All patients received fractionated radiation and chemotherapy with 81% receiving concurrent chemoradiation. Med follow-up from start of therapy was 20 mo (rg 2-168 mo). Median overall survival (OS) was 21 mo (2-yr OS 43%). Median progression-free survival (PFS) was 11 mo (2-yr PFS 26%). Immune marker expression was: PDL1-med 3% (rg 0-58%), PDL2-med 0.03% (rg 0-0.5%), PD1-med 0% (rg 0-76%) and CD3-med 21% (rg 0-78%). PDL2, PD1 and CD3 were not associated with outcomes. However, PDL1 > 8% (chosen as the 75th-percentile) was associated with lower risk of distant metastases (DM) (HR 0.57, 95% CI 0.31-0.98, p = 0.04) and a trend towards longer time to disease progression (HR 0.63, 95% CI 0.37-1.02, p = 0.06). There was no association with local-regional recurrence or OS. On MVA including disease stage, PDL1 > 8% was associated with lower risk of DM (HR 0.59, 95% CI 0.35-0.96, p = 0.03) and overall disease progression (HR 0.53, 95% CI 0.29-0.93), p = 0.02). Conclusions: PDL1 > 8% is associated with lower risk of DM and better PFS in LA-NSCLC. This data suggests that even with traditional cancer therapies, the best outcomes may be seen in patients with higher PDL1 expression.
Identification of HLA-A0201 restricted epitope of novel cancer/testis antigens VCX/Y and generation of antigen-specific T-cell receptor engineered T cells for treatment of solid tumor malignancies.
Authors: Pan Ke , Yee Cassian
Published: February 2018

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Background: To identify HLA-A0201 restricted epitope of novel cancer/testis antigen VCX/Y, generate antigen specific T cells and T-cell receptor (TCR) engineered T cells for adoptive cell therapy (ACT) of solid cancer patients. Methods: Reverse-immunology method was used to identify HLA-A0201 restricted epitope of VCX/Y. The high binding score peptide or whole length of VCX3A mRNA were pulsed or transfected to mature dendritic cells (mDC) from HLA-A0201+ donor and then stimulated autologous naïve T cells. Tetramer guided sorting were performed to purify the epitope specific T cells and CTL clones were generated with limiting dilution. TCR were cloned out from high activity CTL clone and the recombinant of retrovirus vector were constructed to introduce the TCR to allogeneic PBMC to generate the TCR engineered T cells. Results: One peptide which its sequence was shared with all VCX/Y members was identified. Interesting, only CTL clone generated from simulation of VCX3A mRNA transfected DC can recognize naturally processed VCX/Y presented by HLA-A0201+ tumor cells. Cold target inhibition detection confirmed that this VCX/Y peptide was naturally processed and recognized by HLA-A0201+ CTL clone. After infection of retrovirus containing the TCR from high activity of CTL clone, the TCR engineered T cells can recognize HLA-A2+ tumor cells but not normal lung cells. Moreover, these TCR engineered T cells specifically secreted IFN-γ in response to T2 cells pulsed with peptide, as well as HLA-A0201+ and VCX/Y overexpressed tumor cells. Conclusions: VCX/Y peptide we identified is a novel candidate peptide antigen for vaccine or for endogenous adoptive T cell therapy. The correlated high activity TCR gene can generate TCR engineered T cells from patients with anti-tumor activity and offer an alternative adoptive T cell treatment for patients with VCX/Y expressing solid tumor malignancies.
Updated overall survival and safety profile of durvalumab monotherapy in advanced NSCLC.
Authors: Hellmann Matthew D. , Antonia Scott Joseph , Balmanoukian Ani Sarkis , Brahmer Julie R. , Ou Sai-Hong Ignatius , Kim Sang-We , Ahn Myung-Ju , Kim Dong-Wan , Gutierrez Martin , Liu Stephen V. , Schoffski Patrick , Jaeger Dirk , Jamal Rahima , Leach Joseph W. , Jerusalem Guy Heinrich Maria , Lutzky Jose , Nemunaitis John J. , Gu Yu , Abdullah Shaad Essa , Segal Neil Howard
Published: February 2018

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Background: Single-agent durvalumab is being evaluated in patients with advanced squamous and non-squamous NSCLC in an ongoing Phase 1/2 study (NCT01693562). Here we present updated survival and safety data in NSCLC patients. Methods: Treatment-naïve (1L) and previously treated (2L or 3L+) stage IIIB/IV NSCLC patients received durvalumab 10 mg/kg Q2W for up to 12 months. Patients were stratified by tumor PD-L1 expression (Ventana PD-L1 [SP263] Assay [PD-L1 high: ≥25% of tumor cells with membrane staining]), treatment line, and histology. Results: As of 05 September 2017, 304 NSCLC patients received durvalumab monotherapy. Median duration of follow-up was 35.6 (0.3–50.9) months. Investigator-assessed ORR ranged between 23.2% and 30.0% among PD-L1 high patients, and between 3.6% and 8.3% among PD-L1 low/negative patients. Median PFS and median OS were longer in PD-L1 high vs PD-L1 low/negative patients (Table). Any-grade treatment-related AEs (TRAEs) were reported in 57.2% of pts (including fatigue, 17.4%, decreased appetite, 9.2%, diarrhea, 8.9%); in 10.2% of pts these were Grade 3 or 4. TRAEs resulting in treatment discontinuation were reported in 17 patients (5.6%); 1 patient had a Grade 5 TRAE (pneumonia). Conclusions: In this ongoing phase 1 study, OS and safety profile appear encouraging in treatment-naïve and previously treated NSCLC patients, particularly among PD-L1 high patients. Further investigation regarding PD-L1 expression for selection of patients who most likely benefit from durvalumab is needed. Clinical trial information: NCT01693562. [Table: see text]
Systemic immune signature of inflammation in metastatic melanoma (MM) patients treated with ipilimumab (IPI) and carboplatin/paclitaxel (CP).
Authors: Miller Wilson H. , Jamal Rahima , Cocolakis Eftihia , Thebault Pamela , Friedmann Jennifer Elisabeth , Kazemi Shirin , Dionne Jeanne , Cailhier Jean-Francois , Lepage Stephanie , Belanger Karl , Ayoub Jean-Pierre M. , Le HB , Lambert Caroline , El Hajjar Jida , Van Kempen Leon , Spatz Alan , Lapointe Rejean
Published: February 2018

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Background: Only a subset of MM patients benefit from treatment with IPI. In an effort to increase response and identify predictive immune biomarkers, we combined IPI with CP. We here report a significant correlation between overall survival (OS) and biomarkers of a pre-existing inflammatory state. Methods: 30 patients were treated with C (AUC = 6) and P (175mg/m2) every 3 weeks x 5 and IPI (3mg/kg) every 3 weeks x 4. Blood was collected throughout. OS Kaplan–Meier curves were compared by the log-rank test. Cutoff thresholds defining high or low levels of chemokines, B cell populations and PD-1+CD8+ T cell populations were established from the mean value of a given variable from all patients. Neutrophil to Lymphocyte Ratio (NLR) and Systemic Immune Inflammation Index defined as Platelet x Neutrophil to Lymphocyte Ratio (SII) were calculated at baseline and tested for association with OS. SII ≥ 1375 and NLR ≥ 5 were considered as high risk groups. Results: Median OS was 16.2 months, with a 3-year OS of 36.7% for all patients. High levels of CCL3 (HR = 2.79, p = 0.0159), CCL4 (HR = 8.36, p < 0.0001) and CXCL8 (HR = 3.52, p = 0.0037) were associated with worse OS. Advanced B cell differentiation before treatment was associated with worse patient outcomes. High levels of early differentiated Bm2 ( > 57%) were strongly associated with better OS: HR = 0.26 p = 0.004 whereas high eBm5+Bm5 ( > 14%) levels were strongly associated with worse OS: HR = 2.65, p = 0.029. Patients with higher proportions of PD-1+CD8+ T cells in circulation during treatment had poorer OS (HR = 3.84, p = 0.004) at week 10; (HR = 3.53, p = 0.005) at week 13 and (HR = 2.84, p = 0.040) at week 24. High risk SII (HR = 3.30, p = 0.0192) and NLR (HR = 2.367, p = 0.0486) were associated with worse OS. Conclusions: We have identified a significant correlation between pre-existing systemic inflammatory state and a poor response to IPI and CP. Specifically elevated CCL3, CCL4 and CXCL8, baseline B lymphocyte subset skewing, increased CD8+PD-1+ T lymphocytes, increased NLR and SII were all strongly associated to worse OS. Comprehensive immune monitoring provides evidence for new circulating biomarkers predicting outcome in MM patients treated with IPI and CP. Clinical trial information: NCT01676649.
A clinical insight into therapeutic sequence in advanced melanoma.
Authors: Vera Aguilera Jesus , Paludo Jonas , Tschautscher Marcella , Yu Cecilia , Duma Narjust , Markovic Svetomir
Published: February 2018

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Background: The optimal sequence strategy of BRAF/MEK inhibitors, anti-PD- 1/PDL-1 and anti-CTLA-4 in metastatic BRAF-mutated melanoma patients (pts) is unknown and no treatment guidelines exist. Therefore, we report a single-institution experience of different treatment approaches using targeted therapy (TT) and immunotherapy and its impact on outcomes. Methods: BRAF-mutated metastatic melanoma pts treated with TT and immunotherapy from 2012 to2017 were analyzed. Six groups were identified based on treatment strategy. All time-to-event analyses were calculated using the Kaplan-Meier method and Wilcoxon test. Results: Forty-four pts were identified. The median age at diagnosis was 49 years (range 21-73), 54% pts were females and 43% developed brain metastases during disease course. The most common approach strategy was immunotherapy followed by TT, the median duration of treatment was 11 and 19 weeks, respectively. Time-to-next therapy (TTNT) following 1st line treatment was similar in pts treated with TT (median 23 weeks [95% CI: 15-31]) or immunotherapy (median 26 weeks [95% CI: 10-33], p = 0.94). A trend towards better overall survival (OS) was seen in pts who received immunotherapy followed by TT (p = 0.09); patients who received salvage chemotherapy (carboplatin/paclitaxel) had significantly longer OS (median 7 years [95% CI: 3.2-7.08]) ( p = 0.03). Conclusions: No differences in TTNT were seen with immunotherapy, TT or combined (triple therapy) when used as 1st or 2nd line. The significant longer OS benefit with 1st line immunotherapy was only seen in patients who received chemotherapy later in their treatment course. [Table: see text]
Avelumab in patients with previously treated metastatic melanoma: Phase 1b results from the JAVELIN Solid Tumor trial.
Authors: Keilholz Ulrich , Mehnert Janice M. , Bauer Sebastian , Bourgeois Hugues Pierre , Patel Manish R. , Gravenor Donald , Nemunaitis John J. , Taylor Matthew H. , Wyrwicz Lucjan , Lee Keun-Wook , Kasturi Vijay , Chin Kevin M. , von Heydebreck Anja , Gulley James L.
Published: February 2018

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Background: Avelumab is a human anti–PD-L1 IgG1 monoclonal antibody that is approved for the treatment of metastatic Merkel cell carcinoma (US and EU) and advanced urothelial carcinoma progressed on platinum therapy (US). Here, we report phase 1b data for avelumab in a cohort of patients (pts) with previously treated metastatic melanoma. Methods: Pts with unresectable stage IIIC or IV melanoma progressed after ≥1 line of therapy for metastatic disease received avelumab 10 mg/kg IV Q2W until progression, unacceptable toxicity, or withdrawal. Endpoints included objective response rate (ORR), progression-free survival (PFS), overall survival (OS), and adverse events (AEs; NCI CTCAE v4.0). Results: As of Dec 31, 2016, 51 pts were treated and followed for a median of 24.2 mo (range 16.1–31.5). Median age was 64 y (range 31–84). Site of primary tumor was cutaneous (n = 28, 54.9%), ocular (n = 16, 31.4%), mucosal (n = 2, 3.9%), or unknown (n = 5, 9.8%). Pts had received a median of 2 prior lines of therapy for advanced disease (range 0–4), including ipilimumab (n = 26, 51.0%). Confirmed ORR was 21.6% (95% CI 11.3–35.3), with complete response in 7.8% and partial response in 13.7%. In pts with cutaneous melanoma, ORR was 28.6% (95% CI 13.2–48.7). There were no objective responses in pts with ocular melanoma; however, 7 pts (43.8%) had stable disease. In pts with ≤1 (n = 25), 2 (n = 17), or ≥3 (n = 9) prior lines, ORR was 36.0% (95% CI 18.0–57.5), 11.8% (95% CI 1.5–36.4), and 0% (95% CI 0–33.6), respectively. Antitumor activity by PD-L1 status will be presented. Median duration of response was not estimable (NE) (95% CI 2.6 mo–NE). Median PFS was 3.1 mo (95% CI 1.4–6.3) and the 6-mo PFS rate was 39.2% (95% CI 25.2–52.9). Median OS was 18.5 mo (95% CI 9.3–NE) and the 12-mo OS rate was 62.3% (95% CI 46.9–74.4). 39 pts (76.5%) had a treatment-related (TR)AE, most commonly infusion-related reaction (25.5%), fatigue (17.6%), and chills (11.8%). 4 pts (7.8%) had a grade ≥3 TRAE. 5 pts (9.8%) had an immune-related AE; all were grade 1/2. No treatment-related deaths occurred. Conclusions: Avelumab showed durable responses, promising survival outcomes, and an acceptable safety profile in pts with previously treated metastatic melanoma. Clinical trial information: NCT01772004.
Association between objective responses (OR) and overall survival (OS) in patients (pts) with metastatic Merkel cell carcinoma (mMCC) treated with avelumab.
Authors: D'Angelo Sandra P. , Hunger Matthias , Schlichting Michael , Hennessy Meliessa G. , Bharmal Murtuza
Published: February 2018

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Background: Response rates are key efficacy endpoints in oncology trials, yet correlations with OS are not universal across cancer types. MCC is a rare and aggressive skin carcinoma, although it is chemo sensitive with high response rates, responses are of short duration with no demonstrated benefit to OS. This study explores the association between OR and OS in mMCC pts treated with the anti-PD-L1 avelumab. Methods: Efficacy data with ≥ 18 months of follow-up from Part A of the phase II avelumab trial (NCT02155647) were analyzed. Pts had prior progression on or after chemotherapy and received avelumab 10 mg/kg IV Q2W until confirmed progression, unacceptable toxicity, or withdrawal. Tumors were assessed for OR every 6 weeks (RECIST v1.1). Using conditional landmark analyses, OS was compared between pts with and without an independently confirmed OR up to day 47. Only pts alive at the landmark (day 47) are included in this analysis. In a sensitivity analysis, a landmark of 89 days was used. A Cox model including OR as a covariate was fit (unadjusted, controlling for age, visceral disease, and number of previous therapies). Results: 20 pts had an initial OR reported up to day 47, with 7 further pts having an OR between day 48 and day 89, and 2 pts having an OR after day 89, including 10 complete responses (CRs). 5 pts died and 1 withdrew consent before day 47 (all without OR). Survival probabilities 18 months after treatment initiation were 0.900 (95%CI: 0.656; 0.974) for pts with OR up to day 47 vs. 0.262 (95% CI 0.157; 0.378) for pts without OR but alive up to day 47. Similar results were observed for the 89-day landmark. In the unadjusted Cox model, OR was associated with a 94% risk reduction of death (hazard ratio: 0.064 [95% CI: 0.022; 0.181]). The hazard ratio in the adjusted model was 0.052 (95% CI: 0.018; 0.152). Conclusions: Avelumab therapy for previously treated mMCC pts demonstrated a clinically relevant and high survival impact. For avelumab, OR was associated with long-term survival, likely driven by the rapid time to response, substantial tumor reductions including CRs and sustained durable responses not reported in the chemotherapy literature for mMCC. Clinical trial information: NCT02155647.