bluuebunny/crossref_metadata_2025 huggingface dataset:
Intralesional anti-PD1 treatment in patients with metastatic melanoma: The pilot study.
Authors: Samoylenko Igor , Korotkova Olga V. , Zabotina Tatiana , Demidov Lev V.
Published: February 2018

188

Background: Systemic anti-PD1 treatment is major advantage for metastatic melanoma patients over the last five years. Nevertheless up to the moment there are no solid predictive factors discovered. On the other hand immune related adverse events develop quite frequent and sometimes could be dangerous. Methods: Patient with advanced cutaneous melanoma with known BRAF status and at least one superficial lesion (suitable for injection) are eligible. One of two commercially available anti-PD1 drugs (nivolumab or pembrolizumab) is administered intralesionaly to patients every 2 weeks with total daily dose not more than 50 mg (5 ml) using a scale-base approach depending on lesions size. Clinical and immunological outcomes assessed every 12 weeks. For assessing lymphoid infiltrate tumor biopsy performed before treatment and on week 12. To detect a systemic objective response rate of 50% with sensitivity of 80% and error type alfa 5% at least 25 patients to be included. Interim analysis was preplanned when 3 (25%) of events occurred. Results: Among 7 pts OR was detected in 4 (54.7%), PD in 3 (42.9%). Bystander effect was seen in 2 of 4 patients (50%). Among 3 non-responders 2 failed to salvage systemic anti-PD1 treatment and one is now on anti-CTLA4. The most common adverse event was injection site pain, which was self-limiting. Baseline immune phenotype of tumor infiltrating lymphocytes (TILs) was available for 5 of 7 pts included. Baseline percent of TILs, CD3+CD8+ and CD8+CD279+ was higher in responders vs non-responders (3.6±4.08% vs 0.8±0, NS, 75.1±20.6% vs 54.8±1.13, NS; 36.75±18.7% vs 42.8±13.1, NS respectively ). Baseline difference in CD11b+CD28- between responders (7.06±7.768) and non-responders (25.0 ±0.00) was statistically significant. Conclusions: these results demonstrates feasibility, tolerability and potential clinical efficacy of intralesional anti-PD1 in melanoma patients. Further study are needed to determine if unsuccessful intralesional administration could predict failure to systemic treatment as well as TILs phenotype could predict intralesional or systemic anti-PD1 clincal benefit Clinical trial information: MEL004IL.
18-month efficacy and safety update from JAVELIN Merkel 200 part A: A phase II study of avelumab in metastatic Merkel cell carcinoma progressed on chemotherapy.
Authors: D'Angelo Sandra P. , Russell Jeffery Scott , Bhatia Shailender , Hamid Omid , Mehnert Janice M. , Terheyden Patrick , Shih Kent C. , Brownell Isaac , Lebbé Celeste , Lewis Karl D. , Linette Gerald P. , Milella Michele , Hennessy Meliessa , Bajars Marcis , Zwiener Isabella , Nghiem Paul
Published: February 2018

192

Background: Merkel cell carcinoma (MCC) is a rare, aggressive skin cancer. Avelumab, a human anti–PD-L1 monoclonal antibody, is FDA and EMA approved for the treatment of metastatic MCC (mMCC). Here, we report efficacy and safety data for avelumab in patients (pts) with mMCC at ≥18-mo of follow-up. Methods: Pts with mMCC and progression on prior chemotherapy received avelumab 10 mg/kg IV Q2W. Objective response rate (ORR), duration of response (DOR), and progression-free survival (PFS) were evaluated by independent review committee (RECIST v1.1) with tumor assessment every 6 wks; overall survival (OS) and adverse events (AEs; NCI CTCAE v4.0) were also evaluated. Results: As of Mar 24, 2017, 88 pts were treated and followed for a median of 23.0 mo (range 18.7–32.0). Median treatment duration was 17 wks (range 2–132; mean 35 wks ±37). Treatment was ongoing in 15 pts (17%); 7 pts (8%) voluntarily discontinued with continuing responses (5 complete responses [CR] and 2 partial responses). Other reasons for discontinuation were disease progression (n = 42; 48%), death (n = 10; 11%), AE (n = 8; 9%), or consent withdrawal/other (n = 6; 7%). Confirmed ORR of 33% (95% CI 23.3–43.8; CR in 11.4%) remained unchanged from previous analysis and median DOR has not been reached (range 2.8–24.9 mo; 95% CI 18.0–not estimable). Responses were ongoing in 20/29 pts (69%), including 5 pts with > 2 y of follow-up. PFS curve showed a plateau with identical 12- and 18-mo rates of 29% (95% CI 19–39). Median OS was 12.6 mo (95% CI 7.5-19.0) and the 12- and 18-mo OS rates were 51% (95% CI 40–61) and 40% (95% CI 29–50), respectively. 66 pts (75%) had a treatment-related (TR)AE, most commonly ( > 10%) fatigue (25%), infusion-related reaction (15%; all grade 1/2), nausea (11%), and diarrhea (11%); 8 (9%) had a grade ≥3 TRAE. 17 pts (19%) had an immune-related AE, which was grade ≥3 in 4 pts (4.5%). No treatment-related deaths occurred. Conclusions: Updated results demonstrate continued durable antitumor activity of avelumab in pts with mMCC, beyond that expected with cytotoxic chemotherapy. The median OS exceeding 1 y and manageable safety profile of avelumab signify a clinically meaningful benefit in pts with mMCC. Clinical trial information: NCT02155647.
Cemiplimab (REGN2810): A fully human anti-PD-1 monoclonal antibody for patients with unresectable locally advanced or metastatic cutaneous squamous cell carcinoma (CSCC)—Initial safety and efficacy from expansion cohorts (ECs) of phase I study.
Authors: Papadopoulos Kyriakos P , Owonikoko Taofeek Kunle , Johnson Melissa , Brana Irene , Gil Martin Marta , Perez Raymond P. , Moreno Victor , Salama April K. , Calvo Emiliano , Yee Nelson S , Safran Howard , Gonzalez Martin Antonio , Aljumaily Raid , Mahadevan Daruka , Mohan Kosalai Kal , Li Jingjin , Stankevich Elizabeth , Lowy Israel , Fury Matthew G. , Homsi Jade
Published: February 2018

195

Background: There is no established standard of care for unresectable locally advanced or metastatic CSCC. UV-induced DNA damage causes hypermutation in most CSCCs. Therefore, these tumors may be responsive to PD-1 checkpoint blockade. In the dose escalation portion of the phase 1 study of cemiplimab (REGN2810), a durable (19 + months) radiologic complete response was observed in a patient (pt) with metastatic CSCC (ASCO 2015, #3024). Methods: ECs enrolled pts with distantly metastatic (EC 7) and locally advanced (EC 8) CSCC. All patients received cemiplimab 3 mg/kg by intravenous infusion over 30 minutes every 2 weeks for up to 48 weeks. Research biopsies were performed at baseline and Day 29 (and at progression, if possible). Tumor measurements were performed every 8 weeks according to RECIST 1.1 to determine overall response rate (ORR). Data cutoff date was 31 Jan 2017. Results: 26 pts were enrolled (10 in EC 7 and 16 in EC 8): median age, 72.5 y (range, 56–88y); median PS 1 (range, 0–1); 21M:5F; median number of prior systemic therapy regimens, 1 (range, 0–2). Median exposure to cemiplimab was 7 doses (range, 1–22). The most common treatment-related adverse event of any grade was fatigue (19.2%). Each of the following ≥ Grade 3 related AEs occurred once: AST elevation, ALT elevation, arthralgia, and rash. ORR (PR + CR, including unconfirmed) and disease control rate (ORR+SD) were 52% (12/23; 4uPR, 5 PR, 2CR, 1 uCR) and 70% (16/23, including 4SD), respectively. Three patients are not yet evaluable. Median PFS and OS have not been reached, and only one patient has experienced PD during cemiplimab treatment after initial response. Correlative studies are in process, including PD-L1 status and whole exome tumor DNA sequencing. Conclusions: Cemiplimab is well tolerated and produces antitumor activity in patients with advanced CSCC. A pivotal trial of cemiplimab for patients with advanced CSCC is enrolling patients. Clinical trial information: NCT02383212.
Real world treatment patterns of first-line combination therapies among BRAF+ metastatic melanoma patients stratified by tumor burden.
Authors: Ghate Sameer , Tang Jackson , Li Zhiyi , Nakasato Antonio Reis
Published: February 2018

198

Background: For patients (pts) with metastatic melanoma (MM) and BRAF V600 mutation (BRAF+), options for first-line (1L) systemic combination therapy include immunotherapy (IO) or targeted therapy (TT). This study describes real world treatment patterns among BRAF+ MM pts treated with 1L ipilimumab+nivolumab (I+N) or dabrafenib+trametinib (D+T), stratified by tumor burden. Methods: A retrospective observational analysis used Flatiron Health’s electronic health record-derived database from Oct ’15 - Jul ’16. Pts were aged ≥18 years with a MM diagnosis, tested BRAF+ prior to therapy, and treated with ipilimumab+nivolumab (I+N) or dabrafenib+trametinib (D+T) as 1L therapy. Low tumor burden was defined as low/normal LDH (≤ 333 IU/L) and no brain metastasis. High tumor burden was defined as high LDH ( > 333 IU/L) or brain metastasis. Baseline characteristics and treatment patterns were descriptively assessed. Kaplan-Meier (KM) analysis measured time to discontinuation. Results: Among 76 BRAF+ pts, 38% (29) were treated with I+N as 1L, and 62% (47) were treated with D+T as 1L. Of these, 45% (13/29) of I+N vs. 32% (15/47) of D+T had low tumor burden, while 41% (12/29) of I+N vs. 49% (23/47) of D+T had high tumor burden. The two cohorts did not differ by age or gender. Treatment patterns are summarized below. Conclusions: Among pts with low tumor burden, I+N demonstrated shorter time to discontinuation and higher discontinuation rate relative to D+T. Treatment toxicity and progression was the main reason for discontinuation of I+N and D+T, respectively. Among pts with high tumor burden, I+N demonstrated longer time to discontinuation but higher discontinuation rate relative to D+T. Progression was the main reason for discontinuation of both I+N and D+T. [Table: see text]
Tumor-infiltrating lymphocytes to predict survival to anthracycline/taxane-based neoadjuvant chemotherapy in breast cancer.
Authors: Ou Qiyun , Yu Yunfang , Xiao Xiaoyun , Luo Baoming
Published: February 2018

2

Background: Current guidelines have limited information regarding tumour-infiltrating lymphocytes (TILs) were predictive in breast cancer survival treated with neoadjuvant chemotherapy, although some studies suggest a benefit. We quantify the prognostic value of this biomarker for neoadjuvant chemotherapy in patients with breast cancer. Methods: PubMed and EMBASE were searched until March 2017 for studies that investigated the association of TILs with survival for anthracycline/taxane-based neoadjuvant chemotherapy in breast cancer. Overall survival (OS) and disease-free survival (DFS) were combined using random-effects meta-analysis calculated as combined hazard ratio (HR) with 95% credible intervals (CIs). The interaction test was adopted in order to determine the differential effect according to the histology subtypes and TILs subsets. The PROSPERO registry number is CRD42017072133. Results: We identified 16 studies comprising 2,523 patients were entitled. According to histology subtypes analysis indicated that TILs were not prognostic markers for DFS and OS in overall population, but high TILs were related to improved DFS (HR = 0.73, 95% CI, 0.63 – 0.86) and OS (HR = 0.65, 95% CI, 0.51 – 0.83) in triple negative breast cancer (TNBC) patients, and high TILs were also related to better DFS (HR = 0.62, 95% CI, 0.40 – 0.96) in HER2–positive patients. For TILs subsets, our results indicated that high TILs were prognostic markers for DFS (HR = 0.94, 95% CI, 0.89 – 0.98) and OS (HR = 0.74, 95% CI, 0.58 – 0.94). In an ad-hoc analysis, increasing CD8+ lymphocytes were associated with improved DFS (HR = 0.46, 95% CI, 0.33 – 0.65) and OS (HR = 0.41, 95% CI, 0.24 – 0.71), high CD8/FOXP3 ratio was correlated with prolonged DFS (HR = 0.43, 95% CI, 0.29 – 0.63) and OS (HR = 0.45, 95% CI, 0.26 – 0.76), while rich FOXP3+ lymphocytes were associated with reduced DFS (HR = 1.47, 95% CI, 1.01 – 2.15) in overall population. Conclusions: The present results reveal TILs as a predictor of survival to anthracycline/taxane-based neoadjuvant chemotherapy in breast cancer patients. Furthermore, predictive benefit appears to be influenced by breast cancer subtypes and TILs subsets as well.
Microarray to reveal LncRNA profile and the role of lncCUEDC1 in BCSCs.
Authors: Zhang Fengchun , Xu Ying chun , Yan Ning
Published: February 2018

3

Background: Recently, a growing amount of reports have shown that long non-coding RNAs (lncRNAs) are involved in breast cancer development, progression and metastasis. However, the correlation between lncRNAs and breast cancer stem cells (BCSCs) has been poorly explored. Methods: We initially isolated BCSCs from mammosphere-cultured cells; then, microarray analyses were carried out to detect the lncRNA signature in BCSCs. In addition, bioinformatics analyses, including Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), were consulted to explore the functional roles of lncRNAs on BCSCs. Results: A total of 142 aberrantly expressed lncRNAs in BCSCs were identified, and of those lncRNAs, 25 were downregulated and 117 were upregulated compared to non-BCSCs. In addition, some of these lncRNAs were randomly selected and verified by RT-PCR and sanger sequencing. Notably, bioinformatics data showed that the lncRNAs that were detected were largely associated with stemness-related signalling pathways. Additionally, an interacting network between lncRNAs and its mRNAs was constructed to further depict the lncRNA functional activities. Furthermore, we found that lncCUEDC1 negatively regulated phenotype and biological functions of BCSCs in vitro. Conclusions: Our work establishes the lncRNAs signature in BCSCs for the first time. These findings provide us with evidence to explore the functionalities of lncRNAs in BCSCs and indicate that lncCUED1 is a prospective target for BCSCs.
Therapeutic effects of 2B8T2M, a novel fusion of ALT-803, an IL-15 superagonist, with 4 single-chains of anti-CD20 antibody in combination with expanded natural killer cells against rituximab sensitive and resistant Burkitt lymphoma (BL).
Authors: Chu Yaya , Kham Su Nang , Alter Sarah , Jeng Emily , Rhode Peter R. , Barth Matthew John , Lee Dean Anthony , Wong Hing C. , Cairo Mitchell S.
Published: February 2018

32

Background: Patients retreated with rituximab often relapse which limit patient treatment options (Goldman/Cairo, Leukemia, 2013). Our group has successfully expanded functional and active peripheral blood NK cells (exPBNK) to target BL (Chu/Cairo, et al, Can Imm Res, 2015). 2B8T2M was generated by fusing ALT-803, an IL-15 superagonist, to four single-chains of rituximab (Liu/Wong, et al, JBC, 2016). 2B8T2M displayed tri-specific CD20 binding activity, activated NK cells to enhance antibody-dependent cellular cytotoxicity, and induced apoptosis of B-lymphoma cells (Liu/Wong, et al, JBC, 2016). Methods: ALT-803 and 2B8T2M were generously provided by Altor BioScience Corporation. NK expansion, NK receptors expression and cytotoxicity were examined as we previous described (Chu/Cairo, et al, Can Imm Res 2015). IFNg and granzyme B levels were examined by ELISA assays. Equal doses of IgG, Rituximab, ALT-803, Rituximab+ALT-803, obinutuzumab (obinu, generously provided by Christian Klein, PhD from Roche) were used for comparison. Results: 2B8T2M significantly enhanced exPBNK cytotoxicity against rituximab-sensitive Raji cells compared to the controls IgG, Rituximab, ALT-803, Rituximab+ALT-803, obinu (p < 0.001, E:T = 1:1). 2B8T2M also significantly enhanced exPBNK cytotoxicity against rituximab-resistant Raji-2R cells (p < 0.001, E:T = 1:1) and resistant Raji-4RH cells (p < 0.001, E:T = 1:1). Furthermore, 2B8T2M significantly enhanced IFN-g and granzyme B production from exPBNK against Raji, Raji-2R and Raji-4RH compared to IgG (p < 0.001), rituximab (p < 0.001), ALT-803 (p < 0.001), Rituximab+ALT-803 (p < 0.001), and obinutuzumab (p < 0.001). Conclusions: 2B8T2M compared to rituximab, ALT-803 or obinutuzumab significantly enhanced exPBNK in vitro cytotoxicity against rituximab-sensitive and –resistant BL cells. The in vivo functions of 2B8T2M with exPBNK using humanized NSG models are under investigation.
Development of a quantitative PD-L1 assay using laser capture microdissection (LCM)-based reverse phase protein microarray (RPPA) workflow: Implications for precision medicine.
Authors: Pierobon Mariaelena , Baldelli Elisa , Hodge K. Alex , Sereni Maria Isabella , Ludovini Vienna , Bellezza Guido , Crino Lucio , Liotta Lance A. , Petricoin Emanuel
Published: February 2018

35

Background: FDA approved IHC-based companion/complementary assays are routinely used to measure PD-L1 for treatment selection. However, IHC cut-point values vary across platforms and are based on subjective analysis that requires antigen retrieval methods. Even in highly selected populations of PD-L1 “positive” patients, clinical benefits are seen only in subgroups of patients. We tested the feasibility of utilizing LCM and RPPA as a new methodology for quantitative, operator independent measurements of PD-L1 on tumor cells. Methods: PD-L1 quantification by RPPA was compared to IHC on 23 lung cancers (LC). Tumor cells were isolated from the surrounding microenvironment using LCM. The E1L3N clone from Cell Signaling was used to quantify PD-L1 expression by RPPA and IHC. Reproducibility across antibody clones was then assessed on LCM procured tumor epithelia from 10 FFPE LC and 71 snap-frozen ovarian cancers (OC). PD-L1 measurements were compared between the Cell Signaling E1L3N and the Ventana SP-142 clone. A quantitative PD-L1 calibrated assay was developed and assessed with the pilot population data. Results: Of the 23 LC, 5 were PD-L1 positive by IHC. The 5 IHC-positive patients had the greatest level of PD-L1 expression by RPPA indicating that the RPPA platform correlates with IHC. However, RPPA quantification showed a dynamic range ~ 4-fold across IHC negative samples. Correlation between PD-L1 detection with the E1L3N and the SP-142 clone in the OC was significant (R2 = 0.85), indicating great concordance across clones. Results were confirmed in the 10 LC samples (R2 = 0.87). Conclusions: The LCM-RPPA workflow captures PD-L1 expression on a continuous quantitative scale. This quantitative output correlated with IHC although it captures a much broader dynamic range in both IHC negative and positive populations. PD-L1 quantification by LCM-RPPA may be less dependent upon the clone used for the detection than IHC. Because the detection is unconstrained by antigen retrieval issues as well as subjectivity of IHC interpretation, this approach may generate a more accurate cut-point of therapeutic response prediction.
A phase 1b study of the anti-PD-1 monoclonal antibody BGB-A317 (A317) in combination with the PARP inhibitor BGB-290 (290) in advanced solid tumors.
Authors: Friedlander Michael , Meniawy Tarek , Markman Ben , Mileshkin Linda R. , Harnett Paul , Millward Michael , Lundy Joanne , Freimund Alison E. , Norris Christie , Wu John , Paton Virginia , Wang Lai , Gao Bo
Published: February 2018

48

Background: The release of tumor-associated antigens may enhance the response to immunotherapy. BGB-A317, a humanized IgG4 variant monoclonal antibody engineered to have no Fc gamma receptor binding, targets the programmed cell death-1 (PD-1) receptor. It is being developed in solid and hematologic malignancies at a dose of 200 mg IV Q3W. BGB-290, a potent inhibitor of PARP 1/2, is hypothesized to promote neoantigen release that may potentially increase the efficacy of BGB-A317. A phase 1 study identified 60 mg BID as the recommended Phase 2 dose (RP2D) for BGB-290. This study consists of initial dose escalation to determine the maximum-tolerated dose (MTD), safety, pharmacokinetic (PK) profile, and preliminary antitumor activity of the combination, followed by expansion into ovarian, breast, prostate, gastric, bladder, pancreatic and small cell lung cancers. Methods: Cohorts of 6–12 patients with advanced solid tumors were treated in each of 5 planned dose levels (DLs). In DLs 1–3, BGB-290 doses ranged between 20–60 mg PO BID with BGB-A317 2 mg/kg IV Q3W. In DLs 4–5, BGB-290 doses were 40 or 60 mg BID; A317 was given at 200 mg IV Q3W based on PK data from a single agent Phase 1 study. Results: As of 31 March 2017, 43 patients [median age 63 years (34–75)] were treated in DLs 1–5. Three patients experienced four dose-limiting toxicities: grade 2 nausea (DL4), grade 2 nausea and grade 2 vomiting (DL5), and grade 4 autoimmune hepatitis (DL5). MTD was identified as BGB-A317 200 mg IV Q3W + BGB-290 40 mg PO BID. The most common adverse event (AE) considered related to both study drugs was fatigue. Immune-related AEs of Grade ≥3 were elevated alanine aminotransferase/aspartate aminotransferase (n = 3), autoimmune hepatitis (n = 3), and hepatitis (n = 1). Complete or partial response was observed in 11 patients, 4 of whom had confirmed PR or CR. Plasma/serum exposure of BGB-290 and BGB-A317 were consistent with those in single-agent trials. Conclusions: The combination of BGB-A317 and BGB-290 was generally well tolerated in patients with advanced solid tumors. These results support the continuation of this trial with enrollment into the disease-specific cohorts. Clinical trial information: NCT02660034.
IFNγ mRNA signature (IFNγ sig), circulating tumor DNA (ctDNA), and survival in NSCLC or urothelial cancer (UC) treated with durvalumab (D).
Authors: Higgs Brandon W. , Morehouse Chris , Kuziora Michael , Brohawn Philip Z. , Sridhar Sriram , Raja Rajiv , Angra Natasha , Abdullah Shaad Essa , Ranade Koustubh
Published: February 2018

51

Background: Associations between early reduction in plasma ctDNA, pretreatment tumoral IFNγ sig, liver metastases and outcomes, and between tumor mutational burden (TMB) and CD274 (PD-L1) mRNA or IFNγ sig in TCGA were evaluated in NSCLC and UC pts treated with D. Methods: Pts received 10 mg/kg Q2W of D in a Phase 1/2 study in advanced solid tumors. RNAseq measured a 4-gene IFNγ sig; top tertile was IFNγ sig+. Pts with PD-L1 expression (Ventana SP263) ≥ 25% tumor cells in NSCLC or ≥ 25% tumor or immune cells in UC were PD-L1+. 70 genes were assayed for DNA variants (Guardant360) in plasma ctDNA pre/posttreatment. TCGA was used to calculate TMB; ≥ median TMB was high. Results: IFNγ sig+ NSCLC or UC pts had higher response and longer median PFS and OS compared with PD-L1+, PD-L1- and IFNγ sig- pts (Table). Responders showed significant decreases in ctDNA mean variant allele frequency (VAF) posttreatment with D; pts with progressive disease showed increased VAF. Pts with decreased VAF at week 6 had longer median PFS and OS compared with those with VAF increases. VAF changes were not associated with IFNγ sig. NSCLC without liver metastases had higher IFNγ sig (P < 0.001) than pts with liver metastases. In TCGA NSCLC and UC, IFNγ sig+ correlated with high TMB; CD274 mRNA did not. Conclusions: IFNγ sig correlated with outcomes and with TMB in NSCLC and UC. CtDNA VAFs were reduced in NSCLC or UC responders after treatment and correlated with longer survival, suggesting utility as an early indicator of clinical benefit. Clinical trial information: NCT01693562. [Table: see text]
Observational analysis of IDO-1 expression in MEL samples assessed using an RNAscope assay.
Authors: Newton Robert Charles
Published: February 2018

53

Background: IDO-1 catalyzes the degradation of trp to kyn and can promote tumor escape from host immunosurveillance. IDO-1 expression evaluation has potential importance in identification of a population with worse prognosis and is important in the clinical evaluation of IDO-1 inhibitors. We have developed an optimized assay for IDO-1 specific message detection using RNAscope technology (RST) and applied it to an analysis of human melanoma (MEL) samples. Methods: 25 MEL samples were evaluated for IDO-1 by IHC and the samples used to optimize the RST assay. IDO-1 expression could be demonstrated in tumor cells (TC) as well as in immune cells (IC) infiltrating the tumor and in IC in the peritumoral area. A scoring method was developed to classify IDO-1 expression in all three populations. An H-score of 5 or greater was used as an arbitrary cutoff for IDO-1 positivity. An additional 27 MEL samples, collected as part of the ECHO202/KN037 study of epacadostat in combination with pembrolizumab, were evaluated. Results: IHC for IDO-1 showed 24% positive samples for tumor expression. As expected based on enhanced sensitivity, RST identified 56% as positive – 83% of the samples were positive by both assays. With IHC, 60% were positive for intratumoral IC expression of IDO-1 while RST identified 68% as positive for IC expression – 93% of the samples were positive by both methods. Analysis of the samples from the ECHO 202 study showed that 85% were positive for TC IDO-1 expression, while 59% were positive for IC IDO-1 expression with 52% positive for both. Of 22 PD-L1+ samples, 20 were positive for IDO-1 in either TC or IC. Of PD-L1- samples, 4 were positive for IDO-1 in either TC or IC. There was a trend toward higher IDO-1 expression in PD-L1+ tumors. In this data set there was no association of IDO-1 expression with BRAF mutation status, elevated LDH or prior exposure to immune therapies. Conclusions: RST detection has the potential to provide a robust and quantitative assay to evaluate IDO1 expression in TC and IC. Further refinement of this assay and determination of its utility in assessing predictors of patient response in MEL patients undergoing combination therapy with epacadostat and pembrolizumab will take place in ECHO-301/KN252 (NCT02752074).
Reversing immune checkpoint inhibitor resistance with adenoviral IL-24 and p53 tumor suppressor immune gene therapy.
Authors: Chada Sunil , Wiederhold Dora B , Menander Kerstin , Ahn Hyomin , Oh Eonju , Yun Chae-Ok , Sobol Robert E.
Published: February 2018

64

Background: Immune checkpoint inhibitors represent an important advance in cancer therapeutics. However, most cancer patients either do not respond or become resistant to this therapy. Methods: We evaluated the ability of tumor suppressors p53 and IL-24, delivered via adenoviral vectors, to reverse immune checkpoint inhibitor resistance and induce abscopal therapeutic effects in the highly immune therapy resistant murine B16F10 melanoma tumor model. To mimic clinical conditions of immune checkpoint inhibitor resistance, animals with established tumors were treated with anti-PD-1 before intra-tumoral delivery of adenoviral vectors encoding tumor suppressors p53 (Ad-p53) or IL-24 (Ad-IL24). Results: Anti-PD-1 had minimal or no therapeutic efficacy when compared to PBS controls. However, there was a reversal of anti-PD-1 resistance in animals treated with Ad-p53, or Ad-IL24, in combination with anti-PD-1. Evaluation of primary tumor growth using ANOVA confirmed synergistic effects of the combination treatments over either agent used as monotherapy (p = 0.0001 for p53, p = 0.002 for IL-24). We also observed statistically significant decreases in contralateral abscopal tumor growth in animals whose primary tumors were treated with either Ad- p53 or Ad-IL24 (p = 0.046, and p = 0.0021, respectively) or in combination with anti-PD-1 (p = 0.02 p53, and p < 0.0001 for IL-24) as compared to animals treated with anti-PD-1 alone. With respect to survival, combined tumor suppressor + anti-PD-1 therapy resulted in a statistically significant increase in survival compared to: a) tumor suppressor therapy alone (p = 0.01 for either Ad-p53 and or IL-24) and b) anti-PD-1 therapy alone (p < 0.001 for p53, and p = x for IL-24). Conclusions: Overall, these results indicate that IL-24 and p53 tumor suppressor immune gene therapy can reverse immune checkpoint inhibitor resistance in primary tumors, and induce abscopal effects inferring the activation of systemic anti-tumor immune responses that reverse resistance to immune checkpoint inhibitor therapy.
Intratumoral administration of the alpha-Gal glycolipid AGI-134 to induce tumor regression in a mouse model of melanoma.
Authors: Shaw Stephen M. , Wigglesworth Kim , Middleton Jenny , Vainstein Haras Abi , Sorani Ella , Tabakman Rinat , Carmi Levy Irit , Kristian Sascha A.
Published: February 2018

68

Background: AGI-134 is a fully synthetic alpha-Gal glycolipid for intratumoral (i.t.) treatment of solid tumors to induce a patient-specific anti-tumor immune response. AGI-134 recruits pre-existing anti-Gal antibodies to the injected lesion, leading to complement activation and enhanced tumor antigen processing. Using the B16.F10 murine melanoma model, we have previously demonstrated that AGI-134 evokes a robust abscopal anti-tumor effect, with treatment of a primary lesion protecting mice from the growth of distant lesions. In the current study, we investigated the response of injected tumors to AGI-134 administration. Methods: Tumors were induced by s.c. injection of B16.F10 or ovalbumin expressing B16 (B16.OVA) cells into the flank of α1,3galactosyltransferase knock out mice. After reaching a treatable size, the tumors were injected twice, 24 hrs apart, with PBS or 1-1.25 mg AGI-134, and tumor growth monitored for up to 32 days. In addition, to measure the activation of complement after treatment with AGI-134, tumors were processed 2 hours after treatment and the i.t. concentration of complement fragment C5a determined by ELISA. Results: Almost 50% of AGI-134-treated B16.F10 tumors fully regressed vs. 24% in the PBS controls. Moreover, AGI-134 treatment had a significant survival benefit, with 23% of AGI-134-treated mice dying or requiring euthanisia due to tumor mass by Day 27 post treatment vs. 43% in the PBS groups (p < 0.05, Mantel-Cox test). In the B16.OVA model, 67% of the AGI-134 treated tumors fully regressed vs. 0% in PBS-treated mice in two independent assays. Comparison of C5a levels in B16.F10 tumors 2 hrs after AGI-134 or PBS treatment demonstrated that AGI-134 induced significant complement activation within injected tumors. Conclusions: Having previously shown that AGI-134 induces an abscopal effect that protects mice from the development of secondary tumors, we now show that AGI-134 also induces regression of injected tumors: AGI-134-treatment induces the activation of complement within the tumor and leads to complete regression of the tumor in significantly more AGI-134-treated mice than PBS-treated. Clinical trials with AGI-134 are planned for 2018.
Tumor infiltrating lymphocyte recruitment in patients with early stage breast cancer after intramammary IRX-2 cytokine immunotherapy.
Authors: Page David B. , Pucilowska Joanna , Berinstein Neil Lorne , Egan James E.
Published: February 2018

7

Background: The IRX-2 biologic is an injectable immunotherapy containing cytokines derived from stimulated lymphocytes. In preclinical models, IRX-2 activates T-cells, natural killer (NK) cells, macrophages, and dendritic cells, and facilitates maturation of antigen-presenting cells. Stromal tumor-infiltrating lymphocytes (sTILs) are associated with improved outcomes in early stage breast cancer (ESBC), and neoadjuvant IRX-2 increased TILs in a head/neck squamous cell carcinoma trial. We conducted a phase Ib trial to evaluate the feasibility/activity of IRX-2 in ESBC. Methods: Beginning 21 days prior to surgical resection, enrolled operable patients with stage I-III ESBC received the pre-operative IRX-2 regimen consisting of a single low dose of cyclophosphamide (300 mg/m2 to facilitate T-regulatory cell depletion), followed by 10 days of subcutaneous peri-areolar injections of IRX-2 into the affected breast (1 mL × 2 at tumor axis and at 90°). Endpoints were feasibility (primary), sTIL count (secondary), and immune monitoring by flow cytometry and immune transcriptome analysis (Nanostring PanCancer Immune Panel). Results: As of October 2017, 12 patients are enrolled and evaluable. All patients received all planned injections with no treatment-related surgical delays, complications, or grade III/IV toxicities. Treatment was associated with a mean 52% relative increase in sTILs (range –25% to +166%, p = 0.02). RNA expression profiling revealed the greatest increases in Th1 cell signature (mean +28.5%, range -5.3% to +283%), but also increases in CD8+, NK, and DC signatures. PDL1 RNA expression increased in 10/12 patients (mean +131.5%, range –83% to +1112%). In blood, increases in T-cell activation markers (ICOS, HLADR, and CD38) and T-reg depletion were observed at 1-month follow-up. Conclusions: Peri-lymphatic IRX-2 was well tolerated with preliminary evidence of sTIL recruitment, PDL1 upregulation, and peripheral lymphocyte activation. Study enrollment is ongoing, including patients for a pre-neoadjuvant-chemotherapy triple-negative ESBC cohort. Clinical trial information: NCT02950259.
The immunomodulatory activity of bemcentinib (BGB324): A first-in-class selective oral AXL inhibitor in patients with relapsed/refractory acute myeloid leukemia or myelodysplastic syndrome.
Authors: Loges Sonja , Gjertsen Björn Tore , Heuser Michael , Chromik Jörg , Vigil Carlos Enrique , Paschka Peter , Ben-Batalla I , Akyüz Nuray , Micklem David , Brown Anthony , Lorens James , Kebenko Maxim , Janning Melanie , Binder Mascha , Fiedler Walter M. , Cortes Jorge E.
Published: February 2018

70

Background: The tyrosine kinase AXL acts as suppressive immune checkpoint of the innate immune system. AXL signalling leads to immunosuppression and tumour immune escape by downregulating dendritic cell activity, modulating efferocytosis as well as favouring an immunosuppressive chemokine profile and M-MDSC expansion. Therefore it represents an interesting novel immune oncology target. Bemcentinib (BGB324) is a first-in-class, highly selective and orally bioavailable small molecule AXL inhibitor in phase II clinical development. In pre-clinical models, inhibition of AXL signalling with bemcentinib reversed multiple established immune suppressive mechanisms leading to increased infiltration of CTLs, NK and NKT cells and decreased infiltration of M-MDSCs (Wnuk-Lipinska, 2017). We explored safety, PK, efficacy and the effects of treatment with bemcentinib on the T- and B cell repertoire in patients with relapsed AML or MDS. Methods: BGBC003 (NCT02488408) is an open-label, dose finding study of bemcentinib in patients with R/R AML or MDS. Three dose levels have been explored and a loading dose of 400 mg on days one to three followed by 200 mg daily thereafter has been established as safe and recommended phase 2 dose. The TCRß repertoire was quantified by NGS of DNA isolated from PBMNCs using an Illumina MiSeq sequencer. TCRß genes and the IGH repertoire were analysed with BIOMED2-TCRß-A and –B and BIOMED2-FR1/-FR3 primer pools, respectively. Using genomic DNA as template, the amplicons were tagged with Illumina adapters and indices in two consecutive PCR reactions. Demultiplexing and FastQ formated data output was generated by the MiSeq reporter. Analysis of TCRß and IGH data was performed on a Microsoft Cloud using our in-house analysis pipeline Pippa. Results: Early evidence of antileukemic activity was seen. Diversification of the TCR and IGH repertoire could be detected in peripheral blood at day 21 of therapy compared to pre-treatment in three and two out of five matched samples, respectively. Conclusions: Bemcentinib shows preliminary evidence of immune modulation in AML. Clinical trial information: NCT02488408.
Correlation of IL-17 with immune suppression involving MDSC, malnutrition, and prognosis in patients with gastric and colorectal cancer.
Authors: Gonda Kenji , Shibata Masahiko , Ujiie Daisuke , Ashizawa Mai , Kikuchi Tomohiro , Okayama Hirokazu , Fujita Shoutarou , Sakamoto Wataru , Nakajima Takahiro , Endo Hisahito , Saito Motonobu , Monma Tomoyuki , Saze Zenichiro , Ohki Shinji , Kono Koji , Takenoshita Seiichi
Published: February 2018

83

Background: Although a causal relationship linking inflammation and cancer immunity is more widely accepted today, precise cell mechanisms mediating this relationship have not been elucidated. IL-17, a pro-inflammatory cytokine primarily secreted by T helper (Th)17 cells, has previously been associated with inflammatory processes in autoimmune disease. The presence of IL-17 and Th17 cells has been confirmed in various invasive cancers and recently linked to immunosuppression in cancer patients. We investigated systemic inflammation, immune suppression, malnutrition, and prognosis associated with IL-17 in patients with gastric and colorectal cancer. Methods: We measured IL-17 in 106 patients, including 43 with gastric and 63 with colorectal cancer, Production of IL-17 stimulated by PHA was measured by ELISA. MDSC (myeloid-derived suppressor cells), which significantly contribute to immunosuppression, were measured by flow cytometry (CD11b+CD14-CD33+). Neutrophil-to-lymphocyte ratio (NLR) and C-reactive protein (CRP) were used as inflammatory markers. Production of IL-12 and the SI (stimulation index) of lymphocytes’ blastogenic response were used as markers of cell-mediated immune response. Results: Production of IL-17 increased in advanced stages of gastric and colorectal cancer. IL-17 positively correlated with levels of MDSC, serum concentrations of VEGF, NLR, and levels of CRP, and was inversely correlated with IL-12 production, SI, and nutritional markers including prealbumin and retinol binding protein. Patient cohorts were divided into two groups with IL-17 level (540 pg/ml) and OS (overall survival) of patients with stages III and IV gastric or colorectal cancer both significantly worse in patients with high production of IL-17 than in those with low IL-17, although the differences were not significant in patients at stages I and II. Conclusions: The present study suggests that IL-17 may reflect an inflammatory impact on the advancement and progression of cancer, and it may serve as useful marker of immune suppression involving MDSC, malnutrition, and poor prognosis in patients with gastric and colorectal cancer.
Frequency and prognostic role of tumor infiltrating lymphocytes and MMR status in advanced non-colorectal GI malignancies.
Authors: Khan Uqba , Hamza Ameer , Abraham Renny , Khurram Muhammad S , Hadid Tarik H. , Mazzara Paul , Kafri Zyad
Published: February 2018

89

Background: Tumor infiltrating lymphocytes (TILs) and mismatch repair gene mutation (MMR) status are emerging biomarkers in immunotherapy. MMR status and TILs have significant clinical implications with respect to treatment with checkpoint inhibitors. We designed a study to determine the frequency and prognostic utility of TILs and MMR in advanced unresectable non-colorectal GI (NCGI) cancers. Methods: This is a retrospective cohort study of patients who were diagnosed with metastatic or unresectable NCGI cancers between 2009 and 2015 at St. John Hospital and Medical Center. Immunohistochemistry panels were performed on representative tissue sections for MMR testing. TILs were assessed on the hematoxylin and eosin stained slide of the same tissue section. MMR was interpreted as deficient (d) or proficient and TILs were categorized as ≤5 or > 5 per high power field. Descriptive statistics were generated using frequency distributions, medians and means. Kaplan-Meier analysis was performed to determine the impact of TILS and MMR on survival; differences by factor were assessed with the Log_Rank test. Results: We analyzed 132 patients; the mean age at diagnosis was 66.7 years, 62.1% were male. All samples had proficient MMR status. The percentage of patients with TILs ≤ 5 was 46.2. There was no statistically significant difference in median overall survival (OS) by TILs when stratified by stage of tumor. When stratified by type of tumor, median OS by TILs level was significantly different only for hepatocellular cancers (HCC) (≤ 5 TILs, 86 days vs. > 5 TILs 312 days, p = 0.031), table 1. Conclusions: Our study suggests that MMR-d tumors are quite rare in advanced NCGI malignancies. TILs can be present in tumor microenvironment of NCGI malignancies. Though the number of patients in our study was small, there was a statistically significant difference in median OS of patients with HCC when stratified by TILs status.[Table: see text]
Quantitative comparison of PD-L1 immuno-histochemical assays in hepatocellular carcinoma: The Blueprint-HCC study.
Authors: Pinato David James , Mauri Francesco A , Spina Paolo , Cain Owen , Siddique Abdul , Goldin Robert D , Victor Stephane , Pizio Corinna , Akarca Ayse , Boldorini Renzo , Mazzucchelli Luca , Black James M , Shetty Shishir , Marafioti Teresa , Sharma Rohini
Published: February 2018

91

Background: Programmed cell death ligand 1 (PD-L1) expression by immunohistochemistry (IHC) enriches for responses to PD-1/PD-L1 inhibitors, however its role as a predictive biomarker in hepatocellular carcinoma (HCC) is inconclusive, with no consensus on any particular assay. We evaluated the performance of 4 different PD-L1 detection assays previously published in landmark clinical studies. Methods: PD-L1 IHC was performed on 4 serial sections from tissue microarray (TMA) blocks containing 100 archival cases of HCC that included tumour and surrounding non-tumorous tissue. Antibody clones E1LN3, 28-8, 22c3, SP263 were compared on the basis of percentage and intensity of staining in malignant cells (M) to generate an H-score (range 0-300). Immune cells infiltrating (ICI) and at the periphery, in non-tumorous tissue (ICP) were scored on a 4-tier system (0-3). Results: Patients were 76% males, 20% HCV-positive, 64% cirrhotic with a median age of 67 years. Median tumour size was 4 cm, 70% of patients had T1-T2 tumours and 48% were of grade 2. The proportion of PD-L1 positive cases according to M-ICI-ICP pattern was 2-6-2% for E1LN3, 10-18-19% for 28-8, 9-22-18% for 22c3 and 5-14-13% for SP263. Pairwise comparison of M H-scores revealed heterogeneity across antibodies, with highest concordance between E1L3N/SP263 (R2 = 0.95), E1L3N/22c3 (R2 = 0.65), 22c3/SP263 (R2 = 0.66) and increasing discordance for 28-8/22c3 (R2 = 0.44), E1L3N/28-8 (R2 = 0.29), and 28-8/SP263 (R2 = 0.26). Detection of PD-L1-positive immune infiltrates using a semi-quantitative scoring system revealed significantly different scores in pairwise non-parametric comparisons of ICI (p < 0.05) but not ICP (p > 0.05 for chi-square test). Conclusions: In the Blueprint-HCC study we demonstrated that quantification of PD-L1 protein levels in tumour cells, intra-tumoural and peri-tumoural infiltrate is characterised by inter-assay discordance in HCC. This has profound implications in the clinical development of predictive correlates of efficacy to immunotherapy in HCC. Sources of such discordance should be explored.
Preclinical development of tumor-infiltrating lymphocyte therapy for metastatic colorectal cancer.
Authors: Sakellariou-Thompson Donastas , Forget Marie-Andree , Roszik Jason , Jackson Kyle R , Kim Young Uk , Crosby Shadarra , Hurd Mark W , Wu Ji , Jiang Zhi-Qin , Hwu Patrick , Lizee Gregory , Menter David , Kopetz Scott , Overman Michael J. , Haymaker Cara L. , Bernatchez Chantale
Published: February 2018

95

Background: Immunotherapy has become an effective cancer therapy, particularly in the case of checkpoint blockade and adoptive T-cell therapy (ACT). ACT exploits the presence of tumor-infiltrating lymphocytes (TIL) by exponentially expanding their numbers ex vivo and re-infusing them into the patient in an autologous setting. With the effectiveness of TIL therapy already well established in multiple phase II studies in melanoma, there is a push to translate it to other cancers in need of improved therapies. Colorectal cancer (CRC) is a cancer where the presence of TIL has been strongly correlated with increased survival. Metastatic CRC (mCRC) has a poor outcome with median overall survival of less than 3 years. At present anti-PD1 therapy is only active in the small subset of mCRC patients (4%) that are MSI-high. We sought to evaluate the ability to generate and characterize TIL from patients with mCRC to provide a rationale for future TIL therapy in this disease. Methods: To assess the feasibility of utilizing TIL ACT for this patient population, we characterized the immune infiltrate of mCRC, TIL growth from tumor fragments (n = 24), as well as the TIL repertoire (n = 4) and anti-tumor potential in samples with available autologous tumor target. Results: Flow cytometry analysis detected a CD4-rich T-cell infiltration at the tumor site as well as a scarce yet activated CD8+ TIL population. The outgrowth of CD8+ TIL from tumor fragments in media containing IL-2 was potentiated by the addition of an agonistic 4-1BB antibody (Urelumab, BMS). Additionally, the use of a 4-1BB mAb improved the likelihood of growing TIL (63% [14/24] to 83% [20/24]) and increased the total yield of TIL grown. T-cell receptor sequencing showed enrichment in the tumor of CD8+ T-cell clones shared between the blood and tumor, suggesting selective expansion at the tumor site. Using IFNγ ELISPOT, CD8+ TIL from one patient with an MSS tumor were found to be reactive against a peptide derived from mutated TP53R116W restricted to HLA-B*39:01. Conclusions: It is possible to expand CD8+ T cells from microsatellite stable (MSS) mCRC that are able to target tumor antigens. Although preliminary, the initial data suggest the feasibility of TIL therapy for mCRC.
MDSC clinical assay for cancer detection and monitoring in urothelial carcinoma.
Authors: Basu Arnab , Rafatnia Ali , Manoli Hugh , Danenberg Genevieve , Epstein Alan L. , Pinski Jacek K.
Published: February 2018

TPS110

Background: Human Myeloid Derived Suppressor Cells (MDSCs) have gained recognition as a significant population of cells that arise in cancer patients to mediate immune tolerance. Some studies have shown MDSC concentrations to be associated with overall survival and response to therapy in urothelial carcinoma. MDSCs comprise a heterogenous population that are yet to be fully characterized. We have identified a set of biomarkers uniquely expressed by the suppressive human MDSC phenotype and have developed an assay targeted to these markers. MDSCs are potentially a relatively cheap and easily performed screening and monitoring tool in cancers. This study will examine how a proprietary clinical assay works in detecting and monitoring MDSCs in blood and urine samples from patients with or without localized or metastatic bladder cancer. Methods: The pilot research trial will enroll 63 subjects in a 1:1:1 ratio in three groups, normal controls over age 40 without evidence of any malignancy, patients with metastatic disease and localized muscle invasive disease prior to cystectomy. This sample size provides 80% power to detect a 1SD difference in MDSC concentrations in these groups on an F test. Blood and urine samples will be drawn at baseline and at 4 months for comparison. The percent CD33+ HLA-DRlow HIF1a+, CD33+ HLADRlow C/EBPb+, CD11b+ HLA-DRlow HIF1a+, and CD11b+ HLA-DRlow C/EBPb+cells as a fraction of PBMC will be determined. Primary Outcome Measures will be 1. Change in MDSC levels in patients with known localized, muscle-invasive bladder cancer who undergo neoadjuvant and surgical treatment. 2. Change in MDSC level in patients with known metastatic bladder cancer who undergo systemic treatment and 3. The direction and magnitude of the changes compared with radiographically assessed tumor burden. Secondary outcome measures are to assess MDSC level measurements in urine cytology analysis at baseline and after treatment to determine whether the two tests correlate in any of the 3 groups of patients defined in this study. Conclusion: This pilot study will examine if a MDSC clinical assay works in detecting and monitoring MDSCs. Recruitment is open to patients at the USC Cancer center and LAC+USC medical center. Clinical trial information: NCT02735512.