bluuebunny/crossref_metadata_2025 huggingface dataset:
Surrogate end points for overall survival in patients with advanced non-small-cell lung cancer in trials of immunotherapy.
Authors: Ou Qiyun , Yu Yunfang , Li Anlin , Chen Yongjian , Wang Ying , Ren Wei , Li Chenchen , Yao Herui
Published: May 2019

e14122

Background: Our objective was to evaluate progression-free survival (PFS) and objective response rate (ORR) as surrogate end points for overall survival (OS) among patients with advanced non-small-cell lung cancer (NSCLC) in randomized trials of immunotherapy. Methods: Individual patient data (total N = 569 patients) were available from two multicentre, randomized, phase II/III trials (POPLAR and OAK). Analysis of OS, PFS and ORR were computed using the Kaplan-Meier estimates method and compared with the log-rank test, hazard ratios [HRs] and 95% confidence intervals [CIs] were calculated by using the Cox regression model. The surrogacy was evaluated using Spearman’s correlation coefficient (ρ) and coefficient of determination ( R²) between treatment effects on the surrogate end point and OS. We further validated the surrogacy in trial-level analysis with 26 NSCLC immunotherapy trials (total N = 12,094 patients) published prior to June 2018. Results: In the patient-level analyses, patients who achieved a response had better OS compared with nonresponders (HR 0.18; 95% CI, 0.11 to 0.29; P < 0.001). Greater OS was also observed in landmark anlyses at 6-month PFS (HR 0.21; 95% CI 0.11 to 0.39; P < 0.001) and 12-month PFS (HR 0.16; 95% CI 0.11 to 0.24; P < 0.001). PFS correlated with OS at the individual level (Spearman ρ = 0.37; P < 0.001). In the trial-level analysis, PFS moderately correlated with OS (Spearman ρ = 0.64; P < 0.001), and the R² was 0.50 (95% CI 0.39 to 0.64). Conclusions: In NSCLC immunotherapy trials, responders and patients with longer PFS had better OS. ORR and PFS are acceptable surrogates for OS in trials of immunotherapy, but caution must be taken when interpreting efficacy in the absence of OS data.
Incidence and clinical pattern of immune related adverse effects (irAE) due to immune checkpoint inhibitors (ICI).
Authors: Thapa Bicky , Roopkumar Joanna , Kim Ann S , Gervaso Lorenzo , Patil Pradnya Dinkar , Calabrese Cassandra , Khorana Alok A. , Funchain Pauline
Published: May 2019

e14151

Background: ICIs have revolutionized outcomes in many advanced malignancies, however their use is associated with irAEs. Methods: An IRB-approved, retrospective chart review was done using the Cleveland Clinic pharmacy database. Patients were included who received six FDA approved (nivolumab, ipilimumab, pembrolizumab, atezolizumab, avelumab, or durvalumab) ICIs from July 2015 to December 2017. irAEs were identified from review of electronic medical records. Descriptive analysis was done to evaluate the incidence, pattern, and severity of irAEs. Results: A total of 1091 patients received ICI therapy, 651 (59.7%) were male, 958 (87.4%) white, 95 (8.7%) black. Lung cancer (540, 49.5%) comprised the majority of the cohort, followed by melanoma (152, 13.9%), and renal cell carcinoma (121, 11.1%). About 996 (91.3%) received treatment with only one ICI, 85 (7.8%) with 2 ICIs, and only 10 patients received 3 ICIs. A total of 487 (44.63%) patients encountered irAEs, 128 (11.73%) resulting in treatment cessation. Fatigue (152, 13.9%) was the most common, followed by dermatologic irAEs (131, 12%). Endocrine irAEs occurred in 108 (9.89%), GI toxicities, namely diarrhea and colitis, were seen in 92 (8.4%) and hepatotoxicity in 54 (4.94%). Other irAEs including rheumatologic, pneumonitis, renal, and neurological adverse effects were documented in 6.5%, 5.1%, 2.56%, and 2.01% respectively. Rare irAEs such as ocular toxicity, cardiac toxicity, and vasculitis were seen in 0.8%, 0.73%, and 0.54%, respectively. Compared with a pooled analysis from clinical trials (De Velasco G et al. Comprehensive Meta-analysis of Key irAEs from CTLA-4 and PD-1/PD-L1 Inhibitors in Cancer Patients) dermatologic irAEs and fatigue were less frequent; diarrhea/colitis, pneumonitis and rheumatologic irAEs were more frequent. Conclusions: irAEs on ICI therapy are very common. Awareness of the relative frequencies of various irAEs, particularly severe and rare irAEs, in a real-world setting can help improve quality of care for cancer patients receiving ICI. [Table: see text]
Phase Ib study of pembrolizumab and trebananib (angiopoietin-2 inhibitor [Ang-2]): Preliminary analysis of the colorectal cancer (CRC) cohort.
Authors: Rahma Osama E. , Cleary James M. , Schlechter Benjamin L. , Ng Kimmie , Eno Jessica , Stroiney Amanda , Giobbie-Hurder Anita , McDermott David F. , Hodi F. Stephen
Published: May 2019

e14160

Background: Ang-2 is produced by endothelial cells, predominantly in tissues undergoing vascular remodeling and in hypoxic microenvironments. Ang-2 partially suppresses T cell activation by increasing PD-L1 expression and decreasing monocyte activation. Our group demonstrated that high pre-treatment serum Ang-2 is associated with reduced overall survival (OS) in patients treated with PD-1 blockade. Methods: We initiated a phase 1b trial to evaluate the safety, efficacy, and immunological effect of the combination of pembrolizumab and trebananib, an Ang-1/2 neutralizing peptibody, in solid tumors. Treatment consisted of an induction phase of pembrolizumab 200 mg every 3 weeks and trebananib weekly (with an initial run-in dose escalation of 15-30 mg/kg) for 12 weeks followed by pembrolizumab alone for 2 years. Here we present the preliminary data in the fully enrolled CRC expansion cohort. Results: The study enrolled a total of 18 microsatellite stable (MSS) heavily pretreated CRC patients. There were no DLTs and 30mg/kg was deemed to be the MTD. This summary is based on 15 CRC patients treated with 30 mg/kg trebananib (3 in dose escalation, 12 in dose expansion) plus pembrolizumab. Nine patients (60%) were female and the median age was 51 years (43-63). The most common treatment-related adverse events (AEs) were abdominal distension, diarrhea, limb edema, transaminitis, and proteinuria (each reported in 40% of the patients). One grade 3 transaminitis and one grade 4 pneumonitis related to pembrolizumab were reported. As of the date of data retrieval, 11 patients were off treatment and four were continuing with a median follow-up of 6.1 months. The response rate was 7% (1 partial response for 14 months) and the disease control rate was 33% (4 stable disease with CEA trending down). Median time to progression and OS were 2.8 (90% CI: 1.5 to 4.9 months) and 11.4 months (90% CI: 2.6 months to ∞), respectively. Conclusions: The combination of pembrolizumab and trebananib is well tolerated and demonstrated promising activity in heavily pretreated MSS CRC. Paired biopsies and blood were collected for planned correlative immune analyses which will be presented at the meeting. Clinical trial information: NCT03239145.
Characterization of a patient-specific T-cell and tumor cell coculture model of immuno-cytotoxicity in colorectal cancer.
Authors: Li Yanhui , Gunn Nicole Ann , Kua Lindsay , Lee Fiona Yi Xin , Koo Si-Lin , Chua Clarinda Wei Ling , Tan Emile , Toh Ee-Lin , Mathew Ronnie , Lee Ser Yee , Tan Wah Siew , Dasgupta Ramanuj , Tan Iain B.
Published: May 2019

e14178

Background: Colorectal Cancer (CRC) is the third most commonly diagnosed cancer in the world. Yet, CRC has been difficult to treat with immunotherapy, as majority (85%) of CRC are Microsatellite stable (MSS), lowly immunogenic and do not respond well. To investigate immuno-cytotoxicity in MSS cancers, we set up an ex vivo co-culture model using patient-derived tumor epithelial cells and autologous tumor-infiltrating lymphocytes (TILs). Aims: To perform phenotypic and functional characterisation of the model, and show we are able to modulate immuno-cytotoxicity. Methods: Fresh tumor, adjacent normal and/or liver metastasis samples were collected and freshly dissociated from 29 colorectal cancer patients undergoing surgery. TILs and tumor epithelial cells were grown in culture. Weekly immunophenotyping of TILs was performed via flow cytometry and data was analysed to identify temporal changes in proportions of cell types. To modulate cytotoxicity, epithelial cell lines were co-cultured with autologous and allogeneic TILs at various effector:target ratios with and without anti-PD1 (Pembrolizumab). Caspase dye was used to detect apoptosis and measure cell death. Results: TILs from most patients were successfully expanded, with a theoretical potential of up to 10,000 times. CD4+ T cells were selectively expanded, while CD8+ T cells decreased in proportion over time. A select group of patients showed an opposite trend. Cell type proportions in metastases mirrored those of the primary tumour while NK cells expanded more in tumour samples than normal samples. Selection proceeded up to 3 weeks then decreased. In the cytotoxicity experiment, we show that cell death is increased at higher effector:target ratios in the presence of autologous TILs. Addition of Pembrolizumab further modulates cytotoxicity in vitro. Conclusions: We have developed and characterised an autologous T cell and epithelial cell co-culture system to evaluate immuno-cytotoxicity in colorectal cancer. This will allow screening of perturbations for improved immuno-cytotoxicity in colorectal cancer.
Prognostic significance of NLR kinetics in operable triple negative breast cancer.
Authors: Ganta Ranga Raman , Nasaka Srividya
Published: May 2019

e14201

Background: Inflammatory response exacerbates mechanisms linked to tumor growth and dissemination. As an index of systemic inflammatory marker, neutrophil lymphocyte ratio (NLR) may be a predictive biomarker of both prognosis and outcome in several malignancies. However very few reports have addressed the association of change in NLR and outcome in operable breast cancer. We evaluated preoperative NLR and postoperative NLR to assess which one would be predictive of disease outcome in triple negative breast cancer patients. Methods: This study included 67 stage I-III triple negative breast cancer patients diagnosed at HCG Cancer center, between 2013 to 2015. Those patients who underwent upfront surgery were included in the study. Patients who received neoadjuvant chemotherapy and those without adequate medical records were excluded. The NLR was calculated from the differential count by dividing neutrophil percentage with lymphocyte percentage. All preoperative NLRs were calculated from medical records, at the first visit. Postoperative NLR was obtained before first cycle chemotherapy. The NLR was divided into high if ratio is > 3 and low if it is ≤ 3. We evaluated prognostic value of NLR on 3 year DFS. Results: The median preoperative NLR was 2.52 (Range 0.77-8.6). The pre op NLR was high in 19 patients (28%) and low in 48 patients (72%). There was no significant difference between two groups in baseline characteristics. Among the preoperative High and low NLR groups, 3 year DFS is statistically significant. The median postoperative NLR was 2.23 (Range 0.89-8.1). The post operative NLR was high in 7 patients (11%) and low in 60 patients (89%). Among the postoperative High and low NLR groups, 3 year DFS is statistically not significant. The 12 (63%) patients in the high preoperative NLR patients were converted to of the low NLR after surgery. Conclusions: Preoperative NLR correlated with outcome in operable triple negative breast cancer than postoperative NLR. The NLR kinetics might be an index of response to the treatment which needs to be evaluated in prospective studies. Drawbacks of the study: single centre, retrospective study and small sample size.[Table: see text][Table: see text]
Identification of immune microenvironment subtypes of breast cancer in TCGA set: Implications for immunotherapy.
Authors: Liu Chang , Xiao Yi , Tao Zhonghua , Hu Xi-Chun
Published: May 2019

e14205

Background: Checkpoint blockade immunotherapy has been reported to show promising responses in breast cancer. However, only a small quantity of patients benefited from immunotherapy. To understand the underlying molecular mechanisms of immunotherapy and distinguish appropriate patients, we profiled the immune microenvironment landscape of breast cancer. Methods: We analyzed 1092 breast cancer samples from the Cancer Genome Atlas (TCGA) profiled by RNA sequencing. Using 141 immune-related genes, the cohort was divided into 2 subtypes (Immune and non-Immune) by k-means cluster analysis. Immune-related gene signatures representing different immune statuses or immune cells were used to profile Immune Class by k-means cluster analysis, and we further classified them into Active and Exhausted Immune Classes. Results: We identified 40% of breast cancer in TCGA set (433/1092) as Immune Class and further classified them into 2 distinct subtypes, Active and Exhausted Immune Classes (17% and 23%, respectively). Compared with non-Active Immune Class, Active Immune Class was associated with black people and negative hormone receptor (all, P < 0.01). We detected higher proportions of basal-like and HER2-enriched cases within Active Immune Class compared to Exhausted and non-Immune Class (basal-like, 41.0% vs. 11.3% and 12.6%; HER2-enriched, 14.1% vs. 7.1% and 6.5%). In survival analyses, Active Immune Class was significantly correlated with favorable disease-free survival ( P = 0.019) and overall survival ( P = 0.041). Multivariate analyses further identified Active Immune Class as an independent prognostic factor for overall survival ( P = 0.024). Conclusions: Our work classified breast cancer into 3 immune microenvironment subtypes. This novel classification can indicate the prognosis of patients, reveal the underlying immunological mechanisms of tumor progression and may assist immunotherapy decision-making for breast cancer patient. [Table: see text]
Immunotherapy clinical trials suggest predictive immune-related biomarkers are tissue specific.
Authors: Jones Wendell , Ganapathi Ram N. , Ganapathi Mahrukh K
Published: May 2019

e14206

Background: RNA-based immune landscape signatures (RILS) portray the breadth of immune and stromal cell activity in the tumor microenvironment (TME). Their predictive and prognostic capability for patients with solid tumors and treated with various therapies has been reported. Methods: Single and multi-variable survival analysis, multi-variable linear models, publicly available data on clinical trials NCT00796445, NCT01621490, NCT02108652. Results: Using publicly available data, RILS were compared to each other and to tumor mutational burden (TMB) to determine their relative ability to identify responders/non-responders and estimate patient survival in a series of immunotherapy clinical trials. These immunotherapy trials include patients treated with a PD1 inhibitor, PD-L1 inhibitor or MAGE-A3 tumor antigen combined with immunostimulant (MAGE-A3). Although no single RILS/TMB biomarker emerged as a clear indicator of immunotherapy response and survival for all cases, there were certain immune-related signatures that recurred as significant (p < .05) or had near-significance (p < .10) related to a specific cancer or therapy. Immune signatures for T cell trafficking, ratios of PD1 to PD-L1 expression, cytotoxic lymphocytes and B cells were consistently predictive and prognostic in melanoma patients treated with the PD1 inhibitor, nivolumab or MAGE-A3. However, while these same signatures were not predictive/prognostic in bladder and kidney/ureter cancer patients treated with the PD-L1 inhibitor, atezolizumab, biomarkers significantly associated with outcome included TMB, TGFB1, and the ratio of T cell trafficking to M2 tumor associated macrophage activity. Since TMB and various immune signatures generally worked better together than individually in predicting outcome, there is potential for utilization of a combination of biomarkers and immune signatures in the design of effective immunotherapy clinical trials. Conclusions: In summary, our results suggest effective immunotherapy trials may require tissue-specific biomarker development. Further, a combination of immune signatures and biomarkers such as TMB shows markedly increased utility compared to single factor such as TMB, PD-1 or PD-L1 expression alone.
Novel monoclonal antibody therapeutics for metastatic castration resistant prostate cancer.
Authors: Ghosh Maloy , Rodrigues Kavitha Iyer , Maity Sunit , Bhattacharjee Sanghamitra , Manjunath Yogendra , Chakrabarty Subhra Prakash , Dubey Ashvini Kumar , Tiwari Anurag , Murugesan Sathyabalan , Halan Vivek
Published: May 2019

e14222

Background: Therapeutic potential of innate immunity comprising Natural killer cell based targets are beginning to unravel the complexity of immune responses. NK cells recognize and induce cytotoxicity of wide range of target cells, such as, tumor cells without prior antigen sensitization. In this study, we have studied Lectin-like transcript 1 (LLT1), a member of the C-type lectin super family, is expressed on target cells and various immune cells. LLT1 isoform 1, is known to interact with CD161, a critical receptor on NK cells. CD161 is expressed on most of human NK cells, NK-T cells, γδ T cells and so on. Tumors exploit the CD161- LLT1 interaction to evade host defense mechanism (“DO NOT KILL” signal); indicating LLT1 as an attractive immunotherapeutic strategy. Methods: Prostate cancer cell lines and other tumor cell lines were used to evaluate novel anti LLT1 antibodies for therapeutic potential - IFNγ production assays and tumor cell death assays were carried out. In vivo efficacy of these antibodies were established using PC3 xenograft in humanized mouse model (HuNOG-EXL). Results: Human androgen independent prostate cancer cell line, PC3 was studied for LLT1 expression and interactions with immune cells, to understand role of LLT1 in metastatic castration resistant prostate cancer (mCRPC). Overexpression of LLT1 on tumor cells was influenced by cytokines and various TLRs. Inhibition of CD161-LLT1 interaction with novel anti LLT1 antibodies leads to IFNγ production and consequent NK cell mediated cytotoxicity – hall mark of anti-tumor responses. Disruption of LLT1 - CD161 innate immunity axis with anti LLT1 antibody releases the break on NK cell cytotoxicity and hence, established a new therapeutic option. PC3 xenograft on HuNOG mouse revealed in vivo efficacy of LLT1 antibody. Significant tumor growth reduction was observed with specific anti LLT1 antibodies alone and in combination with check point antibodies. Thus, synergistic tumor growth reduction was established by combinatorial application of anti LLT1 antibody and PD1/PDL1 axis inhibitors. Conclusions: PC3 xenograft study and other results point to therapeutic opportunities in metastatic castration resistant prostate cancer, a disease condition which needs improved patient outcomes. The ligation of CD161/LLT1 will serve as a new immuno-oncology pair regulating innate and adaptive immune responses; novel human antibodies against LLT1 described here will bring therapeutic benefit to patients in need.
An innovative combined immunization platform for personalized cancer immunotherapy.
Authors: Matta Jessica , Matta Célia , Thiebault Peter Emilie , Moulaert David , Drillien Robert , Petit-Demouliere Benoit , Sorg Tania , Bou about Ghina , Limacher Jean-Marc
Published: May 2019

e14225

Background: Activity of immune checkpoint inhibitors relies mainly on the presence of an immune response directed against neoantigens resulting from tumor specific mutations. The induction and/or amplification of such an immune response is expected to increase the activity of these therapies. We describe here a novel immunization platform developed for the purpose of personalized cancer immunotherapy. This platform integrates a DNA vector coding for neoantigens, a live modified vaccinia of strain Ankara (MVA) used as a physiologic adjuvant and anti-CTLA-4 as a locally acting early immune checkpoint blocker. Methods: Immune potency was assessed in C57BL6 mice injected subcutaneously three times five days apart with an ovalbumine (OVA) expressing DNA vector (100 µg), either alone or in combination with increasing doses of MVA (up to 2.5x107 plaque forming units, pfu) and increasing doses of anti-CTLA-4 (up to 100 µg). OVA specific immune responses were measured by ELISpot. Anti-tumor efficacy was then investigated with a similar administration scheme in a therapeutic B16F10 mice melanoma model with a DNA vector coding for the B16F10-M30 tumor neoantigen. Results: At an optimal dose of 2.5x106 pfu, MVA significantly improved OVA specific immune response up to 10 times higher as compared to vector alone. Addition of CTLA-4 blockade further increased the magnitude of response, up to 30 times higher than with vector alone. Both MVA and CTLA-4 demonstrated a bell-shaped dose dependent effect. In tumor-bearing animals, 80% experienced durable tumor-free survival when treated with the combination therapy as compared to less than 20% in untreated animals or animals treated with each component independently. Treatment appeared feasible and well-tolerated. Conclusions: Neoantigen coding DNA vector, MVA and CTLA-4 immune checkpoint blockade, when co-administered in immunocompetent C57BL6 mice, acted synergistically to induce a cellular immune response. The same approach translated into a strong anti-tumoral response in an aggressive melanoma model. This combined immunization platform appears as a potential novel way to enhance clinical benefit from current immune checkpoint inhibitors.
Occupational health risk of pressurized intra-peritoneal aerosol chemotherapy (PIPAC) via endoscopical microcatheter system.
Authors: Khosrawipour Tanja , Pigazzi Alessio , Chaudhry Haris , Mikolajczyk Agata , Schubert Justyna , Khosrawipour Veria
Published: May 2019

e14231

Background: Pressurized Intraperitoneal Aerosol Chemotherapy (PIPAC) is becoming an increasingly widespread approach for delivering intra-peritoneal chemotherapy (IPC) by means of a chemoaerosol. In current studies, a flexible and easy-to-handle endoscopic microcatheter (MC) approach has been introduced to replace the conventional micropump used in PIPAC. While this new method has many advantages such as wide availability, cost effectiveness and improved drug distribution, its possible occupational health risks remain unknown. Methods: PIPAC was performed on three post mortem swine using the MC device. According to standard protocol in patients, solutions of doxorubicin and oxaliplatin were aerosolized in a standardized procedure room with laminar flow. Multiple probes of all surgical material including instruments and sutures, surrounding area surfaces and the operating personals’ items such as gloves and gowns were collected. Moreover, the air filter of the operating room was examined for doxorubicin and oxaliplatin concentrations. Results: The majority of probes showed no measurable doxorubicin or oxaliplatin concentrations. No exposure was detected on the operating personals’ gowns as well as the surrounding area surfaces. While a few of the surgical materials as well as the personals’ gloves and the main part of the application system showed low drug exposure, no drug exposure was measurable in the room filter. Conclusions: Our ex-vivo data suggest that PIPAC delivered via MC is safe and does not exhibit any major occupational hazards if the safety protocol is followed. MC offers an improved, easy-to-handle and cost-effective alternative to the micropump used in conventional PIPAC. Furthermore, the MC offers the great advantage of flexibility which allows access even in anatomically difficult situations and reduces the surgical point of entry to only one abdominal trocar placement.
Targeting KDM1A with iadademstat in combination with immunotherapy in an in vivo model of melanoma.
Authors: Maes Tamara , Mascaró Cristina , Sacilotto Natalia , Lufino Michele MP , Ciceri Filippo
Published: May 2019

e14248

Background: By recruiting the immune system against cancer cells, immunotherapy holds promise to achieve long-lasting responses in several malignant conditions. However, a therapeutic response is observed only in a subset of patients. The low mutational burden of “cold” tumors enables them to stay undetected from the host inflammatory response (innate resistance). Also, cancer cells develop a number of coping strategies in response to the selective pressure applied by the treatment with checkpoint inhibitors (recruitment of regulatory cells, defective antigen presentation, immunosuppressant mediators, reduced costimulation, and T cell apoptosis). Hence combinatorial treatments are being considered to tackle the problem of resistances and responsiveness. Iadademstat (ORY-1001) is a selective, orally-available KDM1A inhibitor with low nanomolar potency (Maes et al., 2018). KDM1A is a histone demethylase involved in transcription regulation and key to tumorigenicity in different hematological and solid malignancies. The tolerability and safety of iadademstat has been evaluated in Phase I studies in AML and SCLC and the drug is currently being investigated in combination with SoC in phase II clinical trials in AML and in SCLC. Methods: The combined efficacy of iadademstat administered orally at 10 µg/kg, a dose that does not provoke weight loss or thrombocytopenia, and an anti-Pd1 antibody was evaluated in the mice syngeneic B16F10 melanoma model. Results: Treatment with iadademstat and the checkpoint inhibitor anti-Pd1 monoclonal antibody was well tolerated. By Day 15, treatment with the anti-Pd1 antibody alone or in combo with iadademstat reduced tumor growth by 45% and 65% relative to vehicle controls. Control animals were sacrificed by day 19 due to progression. By day 22, 54% reduction was achieved in the animals treated with the combination compared to the animals treated with the anti-Pd1 antibody alone. Conclusions: This proof-of-concept in vivo experiment highlights the potential of iadademstat in combination with checkpoint inhibitors, prompting further investigation in other oncological indications, characterized by low or no response to the currently approved immune-oncology treatments.
Immune-related adverse events for different age groups using the FDA Adverse Event Reporting System.
Authors: Myint Zin , Qasrawi Ayman , O'Neal Richard , Thomas Chris Z , Arnold Susanne M. , Wang Peng
Published: May 2019

e14256

Background: Understanding immune-related adverse events (irAEs) for different age groups is crucial in the era of immunotherapy. The aim of our study is to assess the irAE profiles in different age groups (≤65 and > 65 years) for drugs that target PD1/PDL1, CTLA4, and their combination. Methods: Data were obtained from the FDA Adverse Event Reporting System database from Jan 2014 to Sep 2018. We extracted the data based on reported irAEs and reviewed 30 different irAEs including, but not limited to colitis, pneumonitis, interstitial lung disease (ILD), myocarditis and pneumonitis. Cases with missing data, another drug reported with the PD1/PDL1 or CTLA4 combinations, and duplicate cases were excluded. We calculated the reporting odds ratio (ROR) for irAE associated with the use of PD1/PDL1, CTLA4 and combination, respectively. Results: A total of 6197 reported irAEs cases were included: 3140 cases from patients ≤65 years and 3057 from > 65 years old. Colitis was the most common irAEs in both groups. The RORs (95% CI) in patients using CTLA4 alone, PD1/PDL1 alone or the combination were 4.17 (3.47-5.02), 0.6 (0.5-0.72), 1.98 (1.64-2.39) for colitis; 0.21 (0.09-0.52), 2.94 (1.97-4.38), 2.2 (1.46-3.3) for diabetic ketoacidosis; 0.29 (0.09-0.92), 16.5 (6.54-41.9), 0.18 (0.04-0.73) for rheumatoid arthritis in ≤65 age group. In > 65 age group, the RORs (95% CI) in patients using CTLA4 alone, PD1/PDL1 alone or the combination were 4.79 (3.49-6.58), 0.56 (0.4-0.78), 2.84 (2.06-4) for hypophysitis; 0.12 (0.06-0.24), 28.53 (18.5-44), 0.17 (0.1-0.29) for interstitial lung disease (ILD); 0.38 (0.19-0.78), 2.87 (2.01-4.1), 2.41 (1.64-3.54) for myocarditis. When we compared all the studied irAEs between age groups (≤65 and > 65) by ROR, the only statistically significant irAE was ILD. The RORs (95%CI) for ILD associated with PD1/PDL1 in age≤65 and > 65 were 11.3 (7.65-16.8) & 28.53 (18.49-44.02). Conclusions: There was no significant difference between irAEs in different age groups except with ILD, which is more commonly seen in the > 65 age group. The ROR combination irAE profiles provide hypothesis generating information regarding association between irAEs and the use of various immune therapies.
Surface programmable activation receptor (SPAR) cell platform for the rapid detection of melanoma-specific biomarkers.
Authors: Lwande Joel , Kittle Joseph D , Williams M. Russell , Liang Shengwen , McQuaid Kyle , Frenchmeyer Melissa , Tang Yuanyuan , Neese Allison , Hua Jiangzhou , McBrairty Charles
Published: May 2019

e14263

Background: We have previously demonstrated that SPAR modified T cells effectively detect bacterial and viral pathogens. SPAR cells are engineered to express a modified T cell receptor (TCR) capable of antibody-directed signal transduction and respond with a Ca2+-mediated production of luminescent signal upon target detection. Here we describe the ability of SPAR cells to rapidly detect the cell surface expression of melanoma biomarkers without a requirement for extensive sample preparation. Methods: SPAR cells expressing both an engineered T cell receptor and the luminescent reporter protein aequorin were developed via the transfection of Jurkat cells with the aequorin expression vector pEF1-Aeq and an engineered TCR complex composed of mouse FcγRI fused to the CD3ζ subunit. The mFcγRI-CD3ζ receptor binds to the Fc region of full-length mouse IgG2 antibodies: SPAR cells are programmed for target detection via the addition of target-specific antibody. The ability of programmed SPAR cells to accurately detect melanoma-specific cell surface biomarkers was evaluated using whole cells from cultured mouse (B16-F10) and human (SK-Mel-28) melanoma cell lines. SPAR cells were programmed by incubation with murine antibodies directed toward the melanoma biomarkers CD133 or TRP1, then mixed with melanoma cells or K562 control cells and evaluated for signal generation. Results: SPAR cells programmed with anti-CD133 or anti-TRP1 antibody produced luminescent signal within minutes when combined with human SK-Mel-28 cells or mouse B16-F10 cells, respectively, known to abundantly express the appropriate biomarker. No signal was generated when programmed SPAR cells were incubated with K562 cells. Further studies also document cytokine production following receptor engagement. Conclusions: SPAR cells can be programmed for the rapid and specific detection of known cell surface cancer biomarkers. The Ca2+-dependent production of luminescent signal and cytokine release in response to TCR engagement suggests SPAR cell activation. Thus, in addition to biomarker detection the SPAR system may ultimately provide predictive insights into the potency of antibody-directed cell therapy.
NSCLC mutation frequency in the central American and Caribbean region.
Authors: Corrales-Rodriguez Luis , Arauz Erik Alexander , Bacon Fonseca Ludwing Alexander , Lanza Mario , Matos Luis Homero , Vargas Carmen
Published: May 2019

e14278

Background: Lung Cancer (LC) constitutes a leading cause of death in the World. Non-small cell lung cancer subtype adenocarcinoma (NSCLC-A) has been associated with epidermal growth factor receptor (EGFR) mutations, which is essential for treatment decisions. The EGFR mutation frequency has been proven to have regional variability in this NSCLC-A population. The current study presents the EGFR mutation incidence and T790M frequency in NSCLC-A cases diagnosed in Central American and Caribbean(CA&C) region. Methods: A cross-sectional evaluation was performed in patients diagnosed with NSCLC-A between 2016 and 2018 in five CA&C countries. Patients were obtained from Reference Centers in Dominican Republic, Nicaragua, Panama, Honduras and Costa Rica through the AstraZeneca testing program. EGFR mutations were evaluated through Real-Time PCR. Results: The research included 972 cases. Mean of age at the diagnosis was 62.79(28.12-90.46) years. Overall distribution of cases by gender was female in 52.37% (509/972) and male in 47.64% (463/972). For the group of antiEGFR TKI naïve patients, tissue samples were processed in 67.83% (601/886). For the group of post antiEGFR TKI patients plasma samples were processed in 90.70% (78/86). For the group of antiEGFR TKI naïve patients, EGFR mutations were present in 26.64% (236 / 886), the mean of age was 63.17 years (28.12-113.47). The frequency of Exon 19 was 53.38(126/236)% and Exon 21 was 27.54%(65/236)% and 11.01% (26/236) were complex mutations. T790M mutations in this treatment naïve population was 2.14%(19/886). For the group of post antiEGFR TKI patients the frequency of T790M mutation was 50.00% (43/86) where 78.57% were obtain from plasma samples. Conclusions: The frequency of NSCLC EGFR mutation positive patients in the CA&C region was similar to that previously described in other Latin American (LA) countries. In the treatment naïve population, tissue samples remain the most important sample to analyze for EGFR mutations. T790M mutations can be seen in treatment naïve population. For patients that progressed to an antiEGFR TKI, T790M mutations were seen in a similar frequency as describe in previous LA countries. For post antiEGFR TKI patients, plasma samples were the preferred method to evaluate T790M mutations with an adequate sensibility.
Development of a PD-L1 multiplex immunofluorescence assay with advanced visual analysis for understanding the tumor microenvironment.
Authors: Downing Sean , Sharma Aditi , Hebert Courtney , Buell Jamie , Wood Douglas
Published: May 2019

e14283

Background: Current PD-L1 Immunohistochemistry (IHC) assays utilizing conventional brightfield, chromogenic 3,3′-Diaminobenzidine (DAB) staining modalities are the norm in both translational and clinical research. With the rise of immuno-oncology, multiplexed immunofluorescence (mIF) has the potential to revolutionize pathology research as it enables the identification of complex cell phenotypes and their potential interactions in the tumor microenvironment (TME). Ultivue has developed a mIF assay for profiling PD-L1, CD8, CD68, pan-Cytokeratin, and Sox10 enabling whole-slide imaging and data analysis in a single workday. Methods: With an expanding number of biomarkers being interrogated, the complexity of the data analysis and visualization tasks increase exponentially. For N markers, a total of 2N phenotypes are possible. To address this problem and reveal the biologically relevant information embedded in the data, we have developed software tools to reduce the complexity, visualize, and quantify spatial distributions of cells across the full spectrum of possible phenotypes. In this study we demonstrate the utility, robustness, and ability to derive meaningful biological insights by validating the accuracy, precision, sensitivity, and specificity of the PD-L1 multiplex immunofluorescence assay. Results: Here we present results that identify all distinct binary phenotypes within cohorts of lung cancer and melanoma samples using qualitative pathology review and an image processing technique called Phenotypic Surface Density Mapping (PSDM). Important features of this quantitative technique are robustness, use of real physical units (e.g. cells/µm2 or intensity/µm2), and data generation in an unbiased fashion to reveal information about every possible phenotype. Conclusions: Applying the pathology review and PSDM to the cohorts of samples confirmed a high level of concordance demonstrating sensitivity and specificity with high degrees of confidence above 85%. Analysis of intra-run, inter-run, and inter-technician runs revealed that cell counts were within a CV less than 15% across all markers. Data presented here indicates the potential for use in a high throughput testing laboratory to aid in collection and interpretation of meaningful biological insights.
Prognostic implications of PD-L1 expression in breast cancer at the protein and mRNA levels.
Authors: Matikas Alexios , Zerdes Ioannis , Lovrot John , Richard François , Sotiriou Christos , Bergh Jonas C. S. , Valachis Antonios , Foukakis Theodoros
Published: May 2019

e14284

Background: Conflicting data have been reported on the prognostic value of PD-L1 expression per immunohistochemistry (IHC) in breast cancer (BC). There is a paucity of data on the role of PD-L1 gene expression (GE). Methods: Medline, Embase, Cochrane Library and Web of Science Core Collection were searched and data were extracted independently by two researchers. Outcomes included pooled PD-L1 positivity in tumor cells, immune cells or both, per subtype and per antibody used; and the prognostic value of PD-L1 positivity for DFS and OS. Heterogeneity was assessed using the Q and I2 statistics. A pooled GE analysis of 39 publicly available transcriptomic datasets was also performed. Results: Of the initial 4184 entries, 38 retrospective studies fulfilled the inclusion criteria. The overall pooled PD-L1 positivity rate in tumor cells was 24%, 33% in immune cells and 25% in both; highest positivity was reported with Dako 28-8 clone. PD-L1 IHC expression in tumor cells was prognostic for shorter DFS (HR = 1.36, 95% CI 1.02 – 1.83, p < 0.04) and OS (HR = 1.66; 95% CI 1.09 – 2.50, p = 0.02); there was significant heterogeneity. PD-L1 IHC expression in immune cells was associated with better DFS (HR = 0.61; 95% CI 0.51 – 0.73, p < 0.001) and OS (HR = 0.53, 95% CI 0.39 – 0.73, p < 0.001) in TNBC. In addition, higher PD-L1 GE predicted better survival in multivariate analysis in the entire population (HR = 0.70, 95% CI 0.60 – 0.82, p < 0.001 for DFS and HR = 0.84, 95% CI 0.75 – 0.93, p = 0.001 for OS) and in basal-like tumors (HR = 0.55, 95% CI 0.38 – 0.80, p = 0.001 for DFS and HR = 0.63, 95% CI 0.50 – 0.79, p < 0.001 for OS), pinteraction 0.124 for DFS and 0.005 for OS. Conclusions: The largest to our knowledge meta-analysis on IHC PD-L1 expression in BC informs on PD-L1 positivity rates and its prognostic value. Standardization is needed prior to routine implementation. PD-L1 GE is a promising prognostic factor, especially in basal-like BC.
Changes in tumor mutational burden in serially biopsied non-small cell lung cancer.
Authors: Smithy James W. , Hwang David H. , Li Yvonne Y. , Spurr Liam , Cherniack Andrew D. , Sholl Lynette M. , Awad Mark M.
Published: May 2019

e14286

Background: High tumor mutational burden (TMB) has been associated with response to checkpoint blockade in non-small cell lung cancer (NSCLC) and other malignancies. However, the degree to which TMB changes over time, across anatomical sites, and with intervening treatment remains unknown. To evaluate TMB changes across time points, we compared TMB in tissue specimens from patients with serially-biopsied NSCLC. Methods: Clinicopathologic characteristicsand changes in TMB were analyzed from patients with NSCLC and more than one biopsy that had undergone targeted next generation sequencing (NGS, OncoPanel) at the Dana-Farber Cancer Institute. Those representing distinct primary tumors by histology and/or discordant mutational profiles were excluded. Results: 134 NSCLC patients with more than one interpretable NGS result were identified; 23 were excluded due to separate primary tumors. Of the 111 remaining patients included in the analysis, the median time between samples was 14 months (range: 0 to 114 months). TMB correlated closely across all matched tumor pairs (Pearson’s r = 0.89), and greater variability in TMB was seen in biopsies taken from different anatomic sites (p = 0.04) compared to biopsies obtained from the same lesion. There was no significant change in median TMB with any intervening therapy, as TMB increased in some patients and decreased in others. Conclusions: In this observational study, TMB correlated closely across tumor pairs. However, these data suggest that sampling from different tumor sites may be associated with greater discrepancies in TMB. It is possible these differences could account for challenges in using TMB as a predictive biomarker for immunotherapy response. Further prospective investigation is needed to inform decisions regarding the need for repeat biopsy in patients starting immunotherapy with remote tissue samples.
BRCA 1/2 mutations in high-grade serous ovarian cancer (HGS-OC) patients of three Latin American countries: Report of 412 cases.
Authors: Molina Juan , Castillo Fernandez Omar Orlando , Lim Maria , Mas Luis , Veliz Ignacio Fernando , Vargas Carmen
Published: May 2019

e14288

Background: Ovarian Cancer (OC) is the most common gynecologic cancer in women in the World and with elevated mortality. High Grade Serous Ovarian Cancer (HGS-OC) is a common histotype and closely related with the presence of BCRA abnormalities. This is to our knowledge the first report of a large series of patients with HGS-OC evaluated in Latin-American countries. The study aim was to present the data obtained from Perú, Costa Rica and Panamá from January 2016 to December 2018. Methods: A cross-sectional evaluation was performed in patients diagnosed between 2016 and 2018 with HGS-OC in three countries in Latin America. The patients were selected from Cancer National Reference Centers in Perú, Panamá and Costa Rica. Germline BRCA 1 / 2 mutations were evaluated through Next Generation Sequencing (NGS) in blood samples. Results: During the period 412 HGS-OC were studied. The mean age at diagnosis was 56.55(SD:12.21) years for all the evaluated patients and 55.42(SD:8.60) years in the mutated cases. Pathogenic HGS-OC mutations in germline BRCA 1 / 2 were diagnosed in 72 cases. Germline BRCA 2 mutations were the most frequent abnormality detected in Panama and in Costa Rica with 70.0 %(7/10) and 77.8%(14/18), respectively. However in Perú, germline mutations in BRCA 1 represents the majority of the mutations with 79.3%(35/44). Variants of uncertain significance (VUS) were detected in 3.64 %(15/412). Positive family history for breast or ovarian cancer were detected in 13.8% of patients with pathogenic mutations(10/72). Specific mutation distribution evidenced in Perú that 6 of 44(13.6%) mutated patients had the mutation in BRCA 1 c.2105dupT; 6 patients of 18(33.3%) had the mutation in BRCA 2 c.5303_5304delTT in Costa Rica and 2 cases had the mutation in BRCA1 c.5186C > A in Panama. The rest of mutations were less common. Conclusions: The frequency of Germline mutations BRCA 1 and BRCA 2 in patients with HSG-OC is similar to the reported in developed nations. However, the different mutation profile between the studied countries could be explained by the Latin-American genetic diversity. Most of the mutations we report were not described in previous studies evaluating breast cancer susceptibility.
Immune checkpoint inhibitor response in tumors with LRP1B variants.
Authors: Zhu Jason , Tucker Matthew D , Kao Chester , Labriola Matthew , Cheris Sachica , Datto Michael B. , Wu Yuan , Healy Patrick , Gupta Santosh , Kirtane Kedar , Gupta Rajan T. , Marin Daniele , Zhang Tian , McNamara Megan Ann , Harrison Michael Roger , George Daniel J. , Armstrong Andrew J.
Published: May 2019

e14291

Background: Low-density lipoprotein receptor-related protein 1B (LRP1B) is a putative tumor suppressor gene spanning > 500 kb on chromosome 2. A melanoma study previously reported enrichment of LRP1B mutations in responders (34%) to immune checkpoint inhibitors (ICIs) compared with non-responders (3%). Deep deletions in LRP1B are frequently observed in non-small cell lung cancer (NSCLC, 12%), head and neck (11%), cervical (9%), bladder (8%), and prostate cancers (8%). This study examines the clinical response to ICIs in a diverse group of tumor types with LRP1B alterations. Methods: We conducted a single center retrospective study inclusive of all patients (pts) with tumors containing any LRP1B variant who were treated with an ICI between 01/2015 and 11/2018. The primary outcome of the study was to describe the clinical outcomes of patients with LRP1B alterations, with respect to radiographic response to therapy, time on therapy, duration of response, and overall survival. Results: 44 pts with an LRP1B variant were treated with an ICI at Duke between 01/2015 and 11/2018. The most common ICIs were pembrolizumab (29/44, 66%) and nivolumab (12/44, 27%). Tumor types included NSCLC (24/44, 55%), renal cell carcinoma (RCC, 5/44, 11%), and prostate cancer (5/44, 11%). 14 (32%) pts had pathogenic variants (PVs) and 30 pts (68%) had variants of uncertain significance (VUS). Among pts with LRP1b PVs, 36% (95%CI 14%-64%) had radiographic responses (5/14, 2 lung, 1 prostate, 1 RCC, 1 endometrial carcinoma), two of whom were MSI high. As a control, the radiographic response rate among patients with a LRP1b VUS was 10% (95%CI 3%-28%). Of those with PVs, 3 pts had stable disease, one of whom has been on CPI therapy for > 22 months and 5 (36%) remain on therapy, with 2 (14%) pts discontinued due to immune toxicities, and 7 (50%) pts discontinued due to disease progression. The median TMB among responders was 12 (2-150.05 mut/Mb) and the median TMB amongst non-responders was 9.5 (6-18 mut/Mb). Conclusions: In a tumor agnostic population of pts harboring LRP1B PVs, 36% of pts have responses to ICI. The majority of responders were MSS and TMB low, which may suggest that PV in LRP1B may independently predict for response to ICI. Prospective and multicenter validation is now needed.
Human model to predict the outcome of cancer vaccine clinical trials.
Authors: Lőrincz Orsolya , Toke Eniko Rita , Somogyi Eszter , Csiszovszki Zsolt , Megyesi Mónika , Pantya Kata , Tóth József , Molnar Levente , Miklós István , Lori Franco , Lisziewicz Julianna
Published: May 2019

e14298

Background: Cancer vaccines activate T-cells with peptides presented by HLA alleles. We hypothesized that peptides binding to multiple HLA alleles of a patient are more likely to induce T-cell responses than epitopes presented by a single HLA. To prove this hypothesis we predicted the outcome of vaccine clinical trials in a Model Population of subjects with complete HLA genotype. Methods: We collected the immune- and objective response rate (IRR, ORR) results from 64 clinical trials conducted in 1,790 patients treated with 42 different vaccines. From these 42 vaccines, we identified personal epitopes (PEPIs) with strong binding affinity to at least 3 HLA alleles of each subject in the Model Population and calculated the proportion of subjects with PEPIs (PEPI Score), multiple PEPIs (MultiPEPI-Score) and PEPIs against multiple vaccine antigens (MultiAg PEPI-Score). Results: Highly significant correlation between the PEPI Score and IRR (p = 0.001) suggests that peptides binding to at least 3 HLA alleles of the subject induce T-cell responses. MultiPEPI-Score correlated with ORR when the tumor of patients expressed the target antigen (p = 0.01). MultiAg PEPI-Score correlated with ORR of clinical trials conducted with vaccines containing multiple antigens (p = 0.01). Conclusions: Our results suggest that (i) a patient’s HLA genotype is the main determinant of vaccine response and (ii) T-cell responses against multiple antigens expressed in the patient’s tumor favors tumor shrinkage. We introduced a novel pre-clinical human model that accurately predicts the outcome of clinical trials, will reduce the risk, cost and time of vaccine development for the prevention and treatment of cancer.