bluuebunny/crossref_metadata_2025 huggingface dataset:
A phase I study of mRNA-2752, a lipid nanoparticle encapsulating mRNAs encoding human OX40L, IL-23, and IL-36γ, for intratumoral (iTu) injection alone and in combination with durvalumab.
Authors: Patel Manish R. , Bauer Todd Michael , Jimeno Antonio , Wang Ding , LoRusso Patricia , Do Khanh Tu , Stemmer Salomon M. , Maurice-Dror Corinne , Geva Ravit , Zacharek Sima , Laino Andressa S , Sun Jing , Frederick Joshua , Zhou Honghong , Randolph William , Cohen Pamela Sarah , Meehan Robert S. , Sullivan Ryan J.
Published: May 2020

3092

Background: mRNA-2752 is a novel mRNA-based therapeutic agent encoding OX40L T cell co-stimulator, IL-23 and IL-36γ pro-inflammatory cytokines. Here we present findings from a first-in-human study of iTu mRNA-2752 in solid tumor patients as monotherapy or in combination with durvalumab (durva). At the time of presentation, data will encompass the monotherapy escalation MTD/RDE along with the supporting translational work, and the available data in combination. Methods: iTu mRNA-2752 was administered every 2 weeks for up to 7 doses as monotherapy or in combination with durva in patients with advanced solid malignancy or lymphoma. Biomarker analyses include measurement of IL-23, IL-36γ and pro-inflammatory cytokine proteins in pre- and post-treatment tumor biopsies and plasma. PD-L1 immunohistochemistry was used to further characterize baseline status and changes to the TME with treatment. Results: As of 20 December 2019, 23 solid tumor patients have been treated either with mRNA-2752 alone (n = 14) or in combination (n = 9) and has been well tolerated with no dose limiting toxicities or related grade 3/4 toxicities. Of the 17 patients evaluated per RECIST and iRECIST, 1 had a PR (iRECIST), 6 had SD, and 10 had PD. The patient with a PR (52% tumor reduction) received 0.5 mg mRNA-2752 with durva, and had aPD-1/L1 naïve squamous-cell bladder carcinoma. Tumor shrinkage was observed in an additional 5 patients in injected and/or uninjected lesions in both monotherapy and combination. Preliminary biomarker data showed increased IL-23 and IL-36γ protein expression after 6-24 hours, and increased levels of downstream cytokines IL-22 and IL-6, respectively. Pro-inflammatory cytokines (e.g. IFN-γ, TNF-α) were also significantly increased at 1 day and 1-week post-treatment. Significant increases in PD-L1 expression predominantly in tumor-associated immune cells were observed after first dose and persisted up to 29 days after treatment. Conclusions: iTu mRNA-2752 given as monotherapy and in combination with durva is tolerable at all dose levels studied, and administration can be associated with tumor shrinkage. Analyses of tumor and plasma biomarkers suggest a sustained immunomodulatory effect of treatment that includes elevated IFN-γ, TNF-α, and PD-L1 levels. These data support the ongoing testing of the mRNA-2752/durva combination in the dose escalation part of the study. Clinical trial information: NCT03739931 .
An in vivo model to evaluate donor-dependent cytokine release in response to single-agent or combination immune-oncology therapies.
Authors: Draheim Kyle , Jiao Jing , Yang Jiwon , Cai Danying , Cheng Mingshan , Keck James G.
Published: May 2020

3114

Background: Although immune-oncology therapies such as checkpoint inhibitor, bi-specific antibody and CAR-T cell therapies are successfully used for cancer therapy, they can have very severe adverse effects such as cytokine release syndrome (CRS). The animal models and in vitro human PBMC assays presently in use do not reliably predict CRS in patients. Currently, the only widely accepted predictors of CRS are cancer burden and therapeutic dose. Despite this, most pre-clinical assays that evaluate CRS do not incorporate cancer cells and the safety of drug combinations has not been widely explored. A predictive assay that identifies patient/cancer/therapy combinations at risk for developing CRS upfront in addition to treatment efficacy would improve the safety of immune-oncology drug development. Methods: We have developed sensitive in vivo humanized mouse models for quantitating CRS that are rapid, reproducible and able to show variation among PBMC donors. The NSG mouse and its derivatives are engrafted with cancer cells and human PBMCs. Mice are then dosed with checkpoint inhibitors or bi-specific antibodies as a single therapy or in combination. Cytokine release is evaluated 2-6 hours post dosing. This assay can be modified to also evaluate efficacy by using luciferase labeled cancer cells and monitoring tumor burden using the Xenogen IVIS imaging system. Results: For all therapy groups, each cytokine tested (including human IFN-γ, IL-2, IL-6, IL-10 and TNF) was upregulated 2-6 hours after drug treatment, but different PBMC donors had various cytokines release levels. Cytokine release levels correlated with a dose response, PBMC engraftment levels and tumor burden. We can demonstrate additive and synergistic cytokine release in the combination treated groups and compare efficacy versus single agents. Our in vivo method was able to determine CRS missed in the in vitro testing method. Conclusions: We have developed a rapid, sensitive and reproducible novel in vivo PBMC humanized mouse model that can differentiate human PBMC donors based on individual safety response to single agent and combination therapeutics of immune checkpoint inhibitors and bispecific T-cell-engaging antibodies. Additionally, this assay can utilize luciferase labelled cell lines to measure treatment efficacy. Using this assay, we can potentially evaluate both cytokine release and efficacy of current immune-oncology therapies as single agents and in combination. This assay has immediate utility in current and future drug development.
Investigating the tumor immune infiltrate for populations that predict immune-related adverse events (irAEs) in patients receiving PD-1 inhibitors.
Authors: Blum Steven Michael , Smith Neal , Sade-Feldman Moshe , Frederick Dennie T. , Jenkins Russell William , Cohen Justine Vanessa , Lawrence Donald P. , Mooradian Meghan , Freedman Christine , Fadden Riley , Rubin Krista M. , Richey Sarah , Flaherty Keith , Wargo Jennifer Ann , Hacohen Nir , Sullivan Ryan J. , Boland Genevieve Marie , Villani Alexandra C
Published: May 2020

3116

Background: The mechanistic relationship between clinical benefit and immune-related adverse events (irAEs) in response to immune checkpoint inhibitors (ICIs) remains unclear, with several clinical studies reporting that irAEs are biomarkers of responses. Single-cell RNA sequencing (scRNAseq) analysis of tumors from patients with advanced melanoma before and after treatment with ICIs have identified immune cells that correlate with response to ICIs. We sought to evaluate if these populations were also associated with irAEs. Methods: A published scRNAseq data set generated with the Smart-Seq2 protocol (Sade-Feldman M, et al. Cell 2018.) was re-analyzed, stratified by two definitions of irAEs: (1) toxicity requiring systemic immunosuppression (prednisone > 10mg/day) or (2) systemic immunosuppression and/or endocrinopathy. Unbiased single-cell analysis was performed, followed by sub-clustering of T cell populations. The percentage of cells in each cluster was determined on a per sample basis. Results: 13,184 immune cells from 39 samples collected from 25 patients were re-analyzed. 27 samples were from patients who did not respond to ICIs, while 12 samples came from responding patients. 21 samples came from patients who required immunosuppression, 5 samples from patients with isolated thyroiditis, and 13 samples from patients who met neither irAE criteria. Unsupervised scRNAseq analyses focused on ICI efficacy re-capitulated published associations between response and populations that included B-cells (p < 0.01) and TCF7 expressing T-cells (p < 0.01). While these cell populations were not associated with either definition of toxicity, we observed a non-Treg CD4 expressing T cell population (0.8-10.5% cells/sample) that positively correlated with either definition of toxicity (p < 0.05) but not efficacy. Conclusions: In a patient cohort with advanced melanoma, tumor-infiltrating immune cell populations associated with response to ICI therapy were not associated with irAEs. This suggests that biomarkers of ICI response may not function as biomarkers of irAEs, and ongoing analysis will seek to validate this result. Understanding the differences between ICI response and irAEs may identify new therapeutic targets for maximizing efficacy while mitigating toxicity.
VCAN accumulation and proteolysis as predictors of T lymphocyte-excluded and permissive tumor microenvironments.
Authors: Emmerich Philip , Matkowskyj Kristina A. , McGregor Stephanie , Kraus Sean , Bischel Kristen , Qyli Tonela , Buehler Darya , Pasch Cheri , Babiarz Christopher , Depke Mitchell , Clipson Linda , Wisinski Kari Braun , Burkard Mark E. , Baschnagel Andrew , Uboha Nataliya Volodymyrivna , Asimakopoulos Fotis , Deming Dustin A.
Published: May 2020

3127

Background: Immune checkpoint inhibitors (ICIs) represent a major advance for treating solid tumors. However, only a minority of patients (pts) benefit from these therapies and markers that predict response have been elusive. Versican (VCAN) is an immunosuppressive proteoglycan in the tumor microenvironment (TME), which releases an immunostimulatory N-terminal fragment versikine (Vkine) when cleaved by ADAMTS proteases. We have demonstrated in colorectal cancers (CRC) that a low VCAN/high Vkine (VCAN proteolytic predominant [VPP]) phenotype correlates with increased tumor-infiltrating CD8+ T lymphocytes (TILs). Here we examine the accumulation of VCAN as a marker of immune exclusion and its proteolysis as a marker of an immune-permissive TME. Methods: Immunohistochemistry for VCAN, Vkine and CD8+ was performed on samples from 1662 pts across breast (BC), CRC, endometrial cancer, pancreatic adenocarcinoma (PDAC), esophageal cancers and neuroendocrine tumors (NETs), across stages of disease (I-IV) and with diverse prior treatments. Stromal intensities of VCAN and Vkine staining quantified in collaboration with blinded surgical pathologists using a 0-3+ scale. 0/1+ were considered “low” for both VCAN and Vkine, whereas 2/3+ were considered “high”. The number of CD8+ TILs were counted using 400x magnification, the equivalent of a high power field (hpf). Results: Across the entire cohort VCAN phenotypes were diverse (VCAN high/Vkine low, 21%; VCAN high/Vkine high, 23%; VCAN low/Vkine low, 29%; VCAN low/Vkine high (VPP), 27%). Consistent with VCAN accumulation as a marker of T cell exclusion, VCAN low cancers had increased TILs compared to VCAN high (4.8 vs 18.3 TILs/hpf, p < 0.001). Low VCAN was identified in 85% esophageal, 79% NET (including small cell lung cancer [SCLC]) 72% endometrial, 47% MSI-H CRCs, 28% triple-negative BC and only 22% MSS CRC, 18% PDAC, 17% ER+ BCs. The VPP subgroup had the highest TILs per hpf across tumors. VPP was identified in 47% of esophageal, 45% endometrial, 41% NETs (including SCLC), 24% MSI-H CRCs, and only 9% MSS CRC, 7% ER+ BCs, 3% triple-negative BCs, and 0% of PDAC (n = 131 PDAC pts). Conclusions: VCAN accumulation correlates with T lymphocyte exclusion, while VCAN proteolysis predicts an immune permissive phenotype. VCAN accumulation and proteolysis are now incorporated into ICI clinical trials as a potential marker of response. Future studies will clarify the role of these biomarkers in predicting benefits of immuno-oncology treatment strategies.
ESR1 mutations provide novel targets for breast cancer immunotherapy.
Authors: Goldberg Jonathan , Qiao Na , Gross Brett , Meric-Bernstam Funda , Guerriero Jennifer , Chen Ken , Philips Anne V. , Peoples George Earl , Alatrash Gheath , Mittendorf Elizabeth A.
Published: May 2020

3135

Background: Estrogen receptor (ER)-positive breast cancer is not considered immunogenic. Standard treatment is endocrine therapy to include aromatase inhibitors (AI). However, constitutively activating mutations in estrogen receptor alpha ( ESR1) emerge with treatment making tumors resistant to AI therapy. While these mutations represent a pathway of resistance, they also represent potential neoepitopes that can be targeted with immunotherapy. Here we characterize the role of ESR1 mutations as novel targets for breast cancer immunotherapy. Methods: Immunogenic epitopes derived from mutated ESR1 (i.e. D538G, Y537S and E380Q) were identified in silico using the Immune Epitope Database and by determining overlapping peptides. In vitro T2 binding assays were used to measure the affinities of these peptides to HLA class-I, specifically HLA-A*0201. Dissociation assays were employed to characterize the stability of the peptide-HLA complex. Peptide-HLA-A*0201 tetramer staining was used to determine the expansion potential of peptide-specific cytotoxic T lymphocytes (CTL) from peripheral blood of healthy females. Cytotoxicity assays were used to determine the ability of peptide-specific CTLs to lyse cells presenting mutated ESR1-derived peptides. Results: We identified 22 nonameric and decameric peptides derived from the most common ESR1 mutations; 10/22 demonstrated high affinity (i.e. IC50 < 500nM) binding to HLA-A*0201. The 3 highest predicted peptides demonstrated low IC50 values (13 nM, 19.5 nM and 56.6 nM), indicating very tight binding to HLA-*0201. In vitro assays confirmed high affinity binding for 10 of the 22 in silico-predicted peptides with an average fold change of 1.52 compared to non-pulsed T2 cells, and a median dissociation half-life of 6.45 hours. Tetramer staining of peptide specific CTLs from normal donor peripheral blood mononuclear cells showed relatively high expansion frequencies, with the highest three frequencies noted for D538G (1.04%), Y537S (0.49%) and V392I (0.27%). Using 4-hour in vitro cytotoxicity assays, in comparison with non-pulsed T2 cells, there was significantly higher lysis of peptide pulsed T2 cells that were cocultured with matching peptide-specific CTL: D538G (67.1 % vs 36.9%, P < 0.001), Y537S (59.5% vs 37.5%, P < 0.01), and E380Q (36.3% vs 7.8%, P < 0.001). Conclusions: These data confirm the immunogenicity of epitopes derived from the most common ESR1 mutations. Further investigation of these peptides as part of novel immunotherapies, to include vaccine strategies is warranted.
Personalized viral-based prime/boost immunotherapy targeting patient-specific or shared neoantigens: Immunogenicity, safety, and efficacy results from two ongoing phase I studies.
Authors: Drake Charles G. , Johnson Melissa Lynne , Spira Alexander I. , Manji Gulam Abbas , Carbone David Paul , Henick Brian S. , Ingham Matthew , Liao Chih-Yi , Roychowdhury Sameek , Kyi Chrisann , Basciano Paul Andrew , Bournazou Eirini , Abhyankar Jesse , Bezawada Ashish , Kounavouth Sonia , Schenk Desiree , Ferguson Andrew R. , Rousseau Raphael F. , Catenacci Daniel V.T.
Published: May 2020

3137

Background: Neoantigens are key targets of a tumor-specific immune response and CD8 T cells targeting neoantigens drive clinical benefit in patients (pts) treated with checkpoint inhibitors. Methods: Two Phase I studies are being conducted to assess the safety, immunogenicity, and early clinical activity of a viral-based neoantigen-targeting prime/boost immunotherapy aimed at maximizing the CD8 T cell response. Both studies use a chimpanzee adenovirus prime followed by increasing doses of repeat boosts with a self-amplifying mRNA in combination with IV nivolumab +/- SC ipilimumab. In the first study, GO-004, patient-specific neoantigens are predicted using Gritstone's EDGE model and incorporated into both prime/boost vectors. In GO-005, shared neoantigens derived from common driver mutations (including several from KRAS) are encoded in off-the-shelf prime/boost vectors. Results: To date, 12 pts have been treated: 6 pts with GEA, NSCLC, or MSS-CRC (GO-004) and 6 pts with NSCLC, MSS-CRC, or PDA (GO-005) with all pts receiving IV nivolumab and 5 pts also receiving SC ipilimumab. Nine pts continue to receive study treatment. No DLTs have been observed. Treatment-related AEs are reversible and include Grade 1/2 fever (7/12), injection site reactions (4/12), fatigue (3/12), diarrhea (2/12), hypotension (2/12), pruritus (2/12), skin reactions (2/12), anorexia (1/12), dyspnea (1/12), hyponatremia (1/12), infusion-related reactions (1/12), myalgia (1/12), and asymptomatic Grade 3 CK elevation (1/12). At the time of analysis, 8 of 12 pts with ≥ 1 radiographic assessment have a best response of stable disease (SD) (3) and progressive disease (PD) (4), and one pt with no evaluable disease at baseline continues on study > 8 months. In GO-005, 1 pt with SD has a 20% reduction in tumor dimensions that correlates with a decrease in ctDNA. In 4 pts in GO-004 analyzed to date, all pts showed substantial neoantigen-specific CD8 T cell responses to multiple neoantigens after priming which increase further in 2 of 3 pts analyzed after subsequent boosts. In GO-005, 1 of 3 pts showed a robust KRAS G12C-specific CD8 T cell response. Induced T cells express IFNg and granzyme B, consistent with an effector response. Conclusions: Taken together, these early data support the tolerability of a viral-based prime/boost immunotherapy, demonstrate marked immunogenicity, and are consistent with potential clinical activity. Additional pts and data at higher dose levels will be presented. Clinical trial information: NCT03639714, NCT03953235 .
Phase I study of 9-ing-41, a small molecule selective glycogen synthase kinase-3 beta (GSK-3β) inhibitor, as a single agent and combined with chemotherapy, in patients with refractory tumors.
Authors: Carneiro Benedito A. , Cavalcante Ludimila , Bastos Bruno R. , Powell Steven Francis , Ma Wen Wee , Sahebjam Solmaz , Harvey Donald , De Souza Andre Luiz , Dhawan Mallika Sachdev , Safran Howard , Giles Francis J. , Munster Pamela N.
Published: May 2020

3507

Background: Overexpression of GSK-3β, a serine/threonine kinase, is associated with advanced stage malignancies, aggressive tumor growth, and chemotherapy resistance. 9-ING-41 is a GSK-3β inhibitor with significant broad spectrum pre-clinical antitumor activity, including chemotherapy-resistant models. This first-in-human study (NCT03678883) is evaluating the safety, pharmacokinetics (PK), and efficacy of 9-ING-41 monotherapy and in combination with chemotherapy in patients (pts) with refractory malignancies. Methods: 9-ING-41 is given intravenously (IV) twice-weekly as a single-agent (21-day cycle) or combined with gemcitabine, gemcitabine/nab-paclitaxel, carboplatin, carboplatin/paclitaxel, doxorubicin, lomustine or irinotecan in patients previously treated with the same chemotherapy. Results: As of Jan 2020, 101 pts were enrolled. Tumor types: 25 pancreatic (PDAC), 14 colorectal (CRC), 10 non-small cell lung cancer (NSCLC), 8 GBM and other gliomas, 7 melanoma, 5 appendiceal, 4 breast (BC), 30 others. Seven single agent dose levels (DL) completed (1, 2, 3.3, 5, 7, 9.3, 12.4 mg/kg) without any 9-ING-41-attributable SAEs. 9-ING-41 attributable AEs include: transient visual change (color perception; n = 29), starting at DL 3 (3.3mg/kg), all G1/2; infusion reactions (n = 14), all G1/2, starting at DL 5 (7mg/kg). 9-ING-41’s mean terminal half-life is 12-20 hrs. Cmax and AUC0-72, are dose proportional with no accumulation. One BRAFV600K-mutated melanoma with > 20 brain metastases, post checkpoint/BRAF/MEK failure has an ongoing CR starting at cycle 2 and sustained after 9 months on treatment. 32 (31%) pts had SD as best response (6 PDAC, 6 CRC, 3 appendiceal, 2 BC, 2 salivary gland, 2 melanoma, 1 Merkel cell, 2 GBM/glioma, 1 RCC, 1 HCC, 1 NSCLC, 1 esophageal, 1 parotid gland, 1 nasopharyngeal, 1 peritoneal, 1 T cell-ALL). 8 pts remained on study treatment > 5 months (1 melanoma, 3 PDAC, 1 appendiceal, 1 GBM, 1 peritoneal, 1 salivary gland) with median treatment duration of 198 days (range 163-261). 32 pts continue to receive 9-ING-41. Conclusions: 9-ING-41 has dose-proportional PK, is well tolerated with significant antitumor activity as monotherapy and in combination with chemotherapy in pts with refractory tumors. 1 ongoing CR was observed in a refractory BRAF-mutated melanoma. A biologically active dose has been reached, although MTD has not been determined. Enrollment is ongoing. Clinical trial information: NCT03678883 .
Utility of circulating tumor DNA (ctDNA) versus tumor tissue clinical sequencing for enrolling patients (pts) with advanced non-colorectal (non-CRC) gastrointestinal (GI) cancer to matched clinical trials: SCRUM-Japan GI-SCREEN and GOZILA combined analysis.
Authors: Ohba Akihiro , Nakamura Yoshiaki , Taniguchi Hiroya , Ikeda Masafumi , Bando Hideaki , Mizuno Nobumasa , Kato Ken , Morizane Chigusa , Nishina Tomohiro , Ueno Makoto , Furuse Junji , Hara Hiroki , Denda Tadamichi , Kawakami Hisato , Esaki Taito , Moriwaki Toshikazu , Yasui Hisateru , Komatsu Yoshito , Yoshino Takayuki , Ohtsu Atsushi
Published: May 2020

3516

Background: We recently reported that clinical assessment of genomic biomarkers using ctDNA had advantages over tumor tissue-based sequencing for enrollment into matched clinical trials across a wide range of GI cancers. Herein we investigated the utility of ctDNA in non-CRC cancers in a SCRUM-Japan GI-SCREEN and GOZILA combined analysis. Methods: In GI-SCREEN, tumor tissue samples of pts with non-CRC were analyzed by a next generation sequencing (NGS)-based assay, Oncomine Comprehensive Assay, since Feb 2015. In GOZILA, plasma samples of non-CRC pts were analyzed by an NGS-based ctDNA assay, Guardant360, since Feb 2018. Results: As of Apr 2019, 2,952 pts in GI-SCREEN and 633 pts in GOZILA were enrolled. Baseline characteristics between the groups were well matched except that GOZILA included more pancreatic (P < 0.0001) and liver cancers (P = 0.016) but fewer gastric cancers (P < 0.0001) and GIST (P = 0.020) than GI-SCREEN. The success rates of the tests were 86.6% in GI-SCREEN and 87.3% in GOZILA (P = 0.649). Median turnaround time (TAT) was 37 days in GI-SCREEN and 12 days in GOZILA (P < 0.0001). The proportion of cases with actionable alterations detected (tissue vs blood; 29.8% vs 46.8%, P < 0.0001) and enrolled into matched clinical trials (4.8% vs 6.5%, P = 0.286) for each group by cancer type are shown in the Table. Pts with upper GI cancers, especially those in GOZILA, were more often enrolled into matched trials; trial enrollment for those with hepatobiliary and pancreatic (HBP) or other cancers was similar regardless of testing method. Median time from GI-SCREEN or GOZILA enrollment to clinical trial enrollment was 5.0 and 1.0 months (mo), respectively (P < 0.0001). Objective response rates (ORR) and progression-free survival (PFS) were not significantly different (tissue vs. blood; ORR: 14.6 vs. 26.3%, P = 0.30: median PFS: 3.3 vs. 2.6 mo, P = 0.71). Conclusions: Clinical sequencing of ctDNA, with its shorter TAT, contributed to rapid enrollment of non-CRC pts into matched clinical trials compared to those tested by tumor tissue sequencing, particularly for those with upper GI cancer, without compromising efficacy. Clinical trial information: UMIN000029315 . [Table: see text]
A phase Ib study of simmitecan (LP) single-agent and in combination with 5-fluorouracil/leucovorin (5-FU/LV) or thalidomide (T) in patients with advanced solid tumor.
Authors: Gong Jifang , Deng Ting , Li Jie , Lu Ming , Li Jian , Ba Yi , Yu Qiuqiong , Shen Lin
Published: May 2020

3523

Background: Simmitecan (LP), a novel 9-substituted lipophilic camptothecin derivative, is a potent inhibitor of topoisomerase I with anti-tumor activity single agent or in combination with other anti-tumor agents, i.e. thalidomide or anti-PD-1 antibody in xenograft models. In this phase Ib study (NCT02870036), we evaluate the safety and anti-tumor effects of LP as monotherapy or in combination therapies in patients (pts) with advanced solid tumors. Methods: This open-label, multi-cohort phase Ib study was conducted at 3 academic centers in China. Eligible pts had advanced cancer without standard treatment options. Prior irinotecan treated failure pts was eligible. In the single agent study, enrolled pts was allocated to received intravenous(iv) 50, 80, or 120 mg/m2 of LP Q2W to determine the safety of LP. After single agent study finished, enrolled pts allocated in LP+5-FU/LV cohorts received 50, 65, or 80 mg/m2 of LP then LV 400 mg/m2 and 5-FU bolus at 400 mg/m2 followed by 5-FU continuous iv of 2400 mg/m2 Q2W; pts in LP+T cohorts received LP 65 mg/m2 Q2W + thalidomide 50mg QD or thalidomide 100mg QD, or LP 80 mg/m2 Q2W + thalidomide 50mg QD. Treatment repeated in 28-day cycles until disease progression or unacceptable toxicity. Results: Between October 2016 to February 2019, 41 pts, median age of 55.1 (range 29-69) years, were enrolled, 13 in LP monotherapy, 10 in LP + 5-FU/LV and 18 in LP + T. Over all, no dose limited toxicity (DLT) occurred. The most common (≥20%) grade 3/4 AE among three regimens was neutropenia (44%, 70% and 89% respectively in LP, LP+5FU/LV and LP+T),and treatment related SAEs were similar, i.e. anemia and febrile neutropenia (11.1% each) in LP, diarrhea (10%) in LP+5-FU/LV, febrile neutropenia (5.6%) in LP+T. Majority of enrolled pts (24/41, 59%) had progressed on prior irinotecan, nevertheless, partial response (PR) was observed in 1 colorectal cancer pt treated with LP 80 mg/m2 + thalidomide 50mg. Disease control rate (DCR, SD≥12 weeks or CR or PR) were 60%, 80% and 61% and median progress free survival were 2.71, 4.17 and 2.76 months respectively in LP, LP + 5-FU/LV and LP+T cohorts. Conclusions: This study showed the safety profiles are manageable, either LP monotherapy or combination therapies, no DLT or safety concern was elicited in combination compared to single agent, and the LP + 5-FU/LV regimen showed higher DCR. A phase II study in pts with metastatic NEC is on-going to explore safety and efficacy of LP + 5-FU/LV regimen combined with toripalimab, an anti-PD-1 antibody. Clinical trial information: 02870036 .
Longitudinal and personalized detection of circulating tumor DNA (ctDNA) for monitoring efficacy of atezolizumab plus bevacizumab in patients with unresectable hepatocellular carcinoma (HCC).
Authors: Hsu Chih-Hung , Lu Shan , Abbas Alex , Guan Yinghui , Zhu Andrew X. , Aleshin Alexey , Lee Kyung-Hun , Lee Michael Sangmin , Mahipal Amit , Ryoo Baek-Yeol , Ding Beiying , Spahn Jessica , Verret Wendy , He Aiwu Ruth , Wang Yulei
Published: May 2020

3531

Background: ctDNA has emerged as a promising biomarker for noninvasive monitoring of treatment response and disease progression in many tumor types. However, the clinical use of ctDNA in patients with HCC has not been established. Here, we evaluated longitudinal and personalized detection of ctDNA for monitoring the treatment response to atezolizumab (atezo) + bevacizumab (bev) in patients with unresectable HCC not previously treated with systemic therapy. Methods: A subset (n = 48) of 104 patients with HCC who enrolled in Arm A of GO30140 (NCT02715531; Phase 1b) and received atezo + bev treatment were included in this study. These patients had 10 CR, 11 PR, 12 SD and 15 PD per IRF-assessed RECIST 1.1. Serial plasma samples were collected at baseline (Cycle [C]1 Day [D]1), during treatment (C2D1, C4D1) and at disease progression. Somatic mutations in individual tumors were identified via whole exome sequencing of archival tumor tissues or fresh biopsies collected before treatment. Personalized ctDNA assays (Signatera 16-plex multiplex PCR next-generation sequencing assay) specific to each patient’s tumor mutational signatures were successfully designed for 47 of 48 patients. Results: At C1D1, a median of 25.7 ng of cell-free DNA was extracted from 2-mL plasma samples. ctDNA was detected in 45 of 47 patients (96%), with a median of 70.6 mean tumor molecules/mL of plasma (MTM/mL) and a median of 1.8% mean variant allele frequency (mean VAF) in plasma. Higher ctDNA levels detected at C1D1 appeared to be associated with increased tumor burden ( P < 0.03). Dynamic changes in ctDNA levels post-treatment were associated with response at C4D1. ctDNA status changed from positive at baseline to negative in 7 of 10 CR (70%), 3 of 11 PR (27%), 1 of 11 SD (9%) and 0 of 11 PD (0%) patients. Longer PFS was observed in patients whose ctDNA became undetectable post-treatment. The median PFS in patients with ctDNA present vs cleared at C4D1 was 6.5 months and not reached, respectively (HR, 12 [1.7-93], log-rank P < 0.00029). Conclusions: Our study showed that Signatera, a personalized and tumor-informed ctDNA assay, could be used as a sensitive method for detecting ctDNA in patients with unresectable HCC. More importantly, our results illustrate the promise of ctDNA as an emerging biomarker that may potentially help to monitor treatment responses and disease progression in patients with HCC.
Comprehensive genomic profiling of 216 Chinese patients with renal cell carcinoma.
Authors: Zhang Congwang , Yang Chunhe , Chen Jie , Xu Xiaohong , Wang Lingyu , Lin Rongbo , Yang Yining
Published: May 2020

3532

Background: Renal cell carcinoma (RCC), a global public health problem, has exhibited a gradual rise in incidence. Unfortunately, the scarcity of effective biomarkers in the clinical became a major limitation of the progress of biological therapies. Therefore, it is imperative to accurately comprehend RCC genomic profiling for exploring its clinical treatment strategies. Methods: Formalin Fixed Paraffin Embedded (FFPE) tumor and matched blood samples of 216 Chinese RCC patients were obtained for next-generation sequencing (NGS)-based 620 cancer genes panel assay and RCC genomic profiling was evaluated. Results: In our Chinese RCC cohort, multiple histological subtypes, encompassing clear cell (96/216, 44.44%), papillary (14/216, 6.48%), chromophobe (2/216, 0.93%), and undefined subtypes (104/216, 48.15%) were included. The top ranked genomic alterations in Chinese RCC patients were VHL (45.83%), PBRM1 (17.1%), BAP1(13.89%), TP53 (10.65%), SETD2 (9.29%), MTOR (8.67%), ARID1A (5.6%), PTEN (5.09%). Interestingly, BAP1, PBRM1 were co-mutated with VHL, and MET, NF2 were mutually exclusive with VHL (all p < 0.05). Of these patients, 87.9% (190/216) of RCC patients had at least one genomic alteration, indicating the potential clinical benefits of targeted therapies. Out of 15 most common canonical pathways, potentially targetable genomic alterations were mainly identified in HIF (45.83%), chromatin remodeling (42.59%), PI3K/AKT (24.07%), DNA damage response (19.44%), RTK-RAS (18.06%), TP53 (15.74%), NOTCH (9.72%), Hippo (9.72%), and WNT (5.56%) pathway. Additionally, HIF pathway was commonly co-altered with chromatin remodeling pathway, and WNT and NOTCH pathways were significantly co-altered with TP53 pathway. Rare mutation types such as CDKN2A-DMRTA1 and HOOK1-ALK were also detected. Across samples, the median TMB was 2.2 (0-18.6) mutations/Mb. Only 2 Chinese RCC patients had TMB-high (> 10 mutations/Mb). Conclusions: Our results displayed the landscape of genomic alterations in 216 Chinese RCC patients. The genomic alterations identified in our study may provide an opportunity to discover potential strategies for targeted and immunotherapy in RCC. Future studies should account for these genomic alterations.
The predictive role of plasma mutant allele fraction to antiangiogenic drugs in patients with mCRC: An expanded analysis of surrogate biomarkers of response to first-line treatment with bevacizumab.
Authors: Martini Giulia , Elez Elena , Mancuso Francesco M , Gomez Maria Auxiliadora , Caratu Ginevra , Matito Judit , Argiles Guillem , Mulet Nuria , Baraibar Iosune , Ros Javier , Garcia Ariadna , Comas Raquel , Ligero Marta , Santos Cristina , Nuciforo Paolo , Dienstmann Rodrigo , Tabernero Josep , Aranda Enrique , Perez-Lopez Raquel , Vivancos Ana
Published: May 2020

3541

Background: So far, no biomarkers of response to anti-angiogenic drugs are available in colorectal cancer (CRC) treatment. Liquid biopsy tracks dynamic mutational changes in CRC patients (pts). RAS mutant allele fraction in plasma (plMAF) is an independent prognostic marker in metastatic CRC (mCRC). We explored the predictive value of plMAF in RAS mutant pts treated in 1st line with chemotherapy +/- bevacizumab (bev). Methods: A multicentric prospective/retrospective analysis was conducted. We collected data from 226 mCRC pts and selected the subset not eligible for metastasis resection with basal plMAF sample evaluable for RAS mutant MAF quantification with digital PCR (BEAMing). Pts were stratified as high (≥ 5.8%) or low ( < 5.8%) plMAF. We investigated associations between clinicopathological variables and progression-free survival (PFS) stratified by plMAF RAS levels using Cox regression models and survival data were calculated by Kaplan-Meier method. Computational analysis of baseline CT scan data extracted 93 radiomics features of all the lesions per patient including 1) 1st class from density histogram distribution and texture analysis by 2) 2nd order and 3) higher order feature classes. The radiomic features distribution between pts with high and low pIMAF was assessed with Student’s t-test analysis. Results: From October 17 to May 19, 63 basal plasma samples were analysed with BEAMing. 42 pts (67.7%) were classified as high and 21 pts (32,3%) as low plMAF. In high plMAF subgroup, a statistically significant longer PFS favouring FOLFOX+bev was observed, compared to FOLFOX alone (10.7 vs 6.9 mts; HR: 0.30; p = 0.002). In low RAS plMAF subgroup, no differences in terms of PFS were observed in either arm (8.9 vs 8.7 mts; HR: 0.7; p = 0.6). Multivariate PFS model showed no association between RAS plMAF and clinicopathological variables, except for high RAS plMAF and treatment benefit with FOLFOX+bev. The CT-radiomics signature, that may translate tumor vascularization, differentiated patients with high vs low pIMAF (p = 0.002). 58 patients (92%) had similar radiomic score; 5 patients with high plMAF (8%) presented very heterogeneous radiomic score distribution. Conclusions: Tumor-borne RAS plMAFs may constitute a potential predictive biomarker of efficacy for anti-angiogenic drugs in mCRC. Next steps will include the identification of -histological, transcriptomic and radiomic- surrogate biomarkers of response that reflect tumor irrigational status.
First-in-human dose-escalation study of anti-EGFR ADC MRG003 in patients with relapsed/refractory solid tumors.
Authors: Xu Rui-hua , Qiu Miao-Zhen , Zhang Yang , Wei Xiao-Li , Hu Chaohong
Published: May 2020

3550

Background: MRG003 is a novel antibody drug conjugate (ADC) composed of a fully human anti-EGFR IgG1 monoclonal antibody conjugated to a microtubule disrupting agent monomethyl auristatin E (MMAE). MRG003 is presently being tested in an ongoing phase I study for safety, pharmacokinetics, and preliminary antitumor activity in patients (pts) with solid tumors (CTR20180310). Methods: In the phase I dose escalation study of a traditional (3+3) design, pts with relapsed or refractory cancers received single agent MRG003 once every 3 weeks (Q3W) for a maximum of 8 treatment cycles. The starting dose of MRG003 is 0.1 mg/kg, followed by 0.3, 0.6, 1.0, 1.5, 2.0, 2.5, and 3.0 mg/kg. Observations included adverse events (AEs), dose-limiting toxicity (DLT), and antitumor activity which is assessed every two cycles. Results: A total of twenty-two pts with colorectal (CRC, n = 15), nasopharyngeal (NPC, n = 3), head and neck (H&N, n = 2), esophageal (EC, n = 1), and duodenal (DC, n = 1) cancer were enrolled in the dose escalation. The median age of pts was 56.5 years. The MTD identified was 2.5 mg/kg. Commonly observed adverse events were anemia (50%), AST increase (41%), decreased appetite (41%), rash (36%), pruritus (36%), asthenia (36%), and proteinuria (32%). Majority of AEs were mild to moderate in severity. EGFR expression in patients’ tumor samples was determined retrospectively by a validated IHC method in a central laboratory. Nine out of 22 pts tested were EGFR positive. Among these 9 EGFR positive pts, one with NPC in the 2.5 mg/kg cohort had partial response, four had stable disease (one with H&N in the 1.5 mg/kg, one each with NPC and H&N in the 2.0 mg/kg, and one with EC in the 2.5 mg/kg cohorts). The disease control rate (DCR) at doses ≥1.5 mg/kg was 100% for the EGFR positive pts. Conclusions: The dose escalation study of MRG003 showed manageable safety profiles and encouraging preliminary antitumor activity in pts with EGFR-positive solid tumors. MRG003 is currently being evaluated as a single agent in phase I dose expansion cohorts to further assess safety, PK, and antitumor activity. Clinical trial information: CTR20180310 .
First-in-human phase I study of ALT-P7, a HER2-targeting antibody-drug conjugate in patients with HER2-positive advanced breast cancer.
Authors: Park Yeon Hee , Ahn Hee Kyung , Kim Ji-Yeon , Ahn Jin Seok , Im Young-Hyuck , Kim Seol-Hee , Lee Sunbae , CHUNG HYE-SHIN , Park Soon Jae
Published: May 2020

3551

Background: ALT-P7 is an antibody-drug conjugate, in which two molecules of monomethyl auristatin E (MMAE) are site-specifically conjugated to a cysteine-containing peptide motif of trastuzumab variant. This is the first-in-human study evaluating safety and pharmacokinetics of ALT-P7 in patients with HER2-positive advanced breast cancer. Methods: This was an open label, 3+3 dose-escalation phase 1 trial. Eligible patients were with HER2-positive advanced breast cancer progressive to at least two kinds of prior anti-HER2 treatment. ALT-P7 at doses from 0.3mg/kg to 4.8mg/kg were intravenously administered once every 3 weeks. Dose limiting toxicities were evaluated over the first cycle of 21 days. Primary objective was to define the maximum tolerated dose. Results: ALT-P7 were administered in 27 patients (n=4 at 0.3mg/kg, n=3 at each of 0.6, 1.2, 2.4, 3.6, 4.2, 4.5 mg/kg, n=5 at 4.8mg/kg) between Jan 2018 and Feb 2020. The median number of previous line of systemic therapy was six, including median four prior anti-HER2 agents. The most common grade (G) 3/4 adverse event (AE) was neutropenia (n=4). The other common drug-related AEs of all grade were myalgia (n=9), fatigue (n=7), sensory neuropathy (n=6), alopecia (n=6), pruritus (n=6), and neutropenia (n=6). Dose limiting toxicities(DLTs) were observed in three patients at 4.8mg/kg (G4 febrile neutropenia, G4 thrombocytopenia, G4 hyperbilirubinemia, G3 myalgia, G4 hyponatremia). No DLTs have been observed up to 4.2mg/kg, and safety evaluation at 4.5mg/kg is currently ongoing. Toxicokinetic analysis for total antibody, drug-conjugated antibody, and free payload suggested that there were no accumulation of ALT-P7 upon repeated injection. In 22 patients with response evaluation, disease control rate at 6 weeks of ALT-P7 treatment was 77.3%(17/22) and partial response was achieved in two out of fifteen patients with measurable lesion. The median PFS at doses from 2.4 to 4.8mg/kg was 6.2 months (95% CI 2.5-9.9 months). Conclusions: ALT-P7 was well tolerated to a dose of 4.2mg/kg in heavily pretreated HER2-positive advanced breast cancer. DLTs were observed at 4.8mg/kg, and 4.5mg/kg is under evaluation. The observed clinical activity warrants further evaluation in a phase 2 trial. Clinical trial information: NCT03281824. Clinical trial information: NCT03281824 .
Anaplastic lymphoma kinase (ALK) partners identified by next-generation sequencing in Chinese patients with solid tumors.
Authors: Yang Sheng , Gong Fuyu , Wang Guoqiang , He Xiaohui
Published: May 2020

3555

Background: Anaplastic lymphoma kinase (ALK) rearrangement is a validated therapeutic driver gene in non-small cell lung cancer (NSCLC). More than 30 different fusion partner genes of ALK in NSCLC have been reportedand most of these ALK fusions respond well to ALK inhibitors crizotinib. With the development of next-generation sequencing (NGS), more novel partners for ALK rearrangement have been identified. Here, we aimed to report the landscape of ALK rearrangement in Chinese patients with solid tumors. Methods: Tissue or blood samples were subjected to NGS in a College of American Pathologists-certified and Clinical Laboratory Improvement Amendments-accredited lab for ALK arrangement. Results: In total, we profiled more than 40,000 patients, among which 72 cases with 52 ALK fusion partner, harboring 17 reported partners and 35 novel partners. The average ALK rearrangement patients' age was 53 years (range, 17-76 years). Among all the ALK fusion cases (n = 72), lung cancer were the largest proportion with 77.8% (n = 56), colorectal cancer accounted for, 5.5% (n = 4), liver cancer accounted for 4.2% (n = 3), biliary cancer, melanoma, carcinosarcoma and inflammatory myofibroblastic tumor accounted for 2.8% (n = 2) respectively, and only one case (n = 1) was malignant peritoneal mesothelioma. The most common ALK fusion partners were KIF5B (n = 6), DCTN1 (n = 5) and STRN (n = 5). In 38 cases, 35 novel ALK fusion partners were discovered. The novel CLIP4-ALK, EHBP1-ALK, PLB1-ALK occurred twice in 6 patients, which were two lung cancer patients with CLIP4-ALK fusion, two lung cancer patients with PLB1-ALK fusion, one hepatic cellular cancer patients with EHBP1-ALK, and one melanoma patients with EHBP1-ALK. There were two special lung cancer cases with two ALK fusions. One case detected the novel LRIG1-ALK fusion and novel PLB1-ALK fusion, the other case detected novel GLI3-ALK fusion and reported HIP1-ALK fusion. Conclusions: Novel ALK fusions are detected in patients with not only NSCLC but also other solid tumors. NGS fusion assay is an optional method for screening novel fusions.
Comparison of the clinical interpretation of high-dimensional molecular data by two molecular tumor boards.
Authors: Rieke Damian Tobias , de Bortoli Till , Horak Peter , Lamping Mario , Jelas Ivan , Rüter Gina , Berger Johannes , Glimm Hanno , Brors Benedikt , Stenzinger Albrecht , Fröhling Stefan , Keilholz Ulrich
Published: May 2020

3564

Background: The clinical interpretation of molecular data is a bottleneck of precision oncology. High-dimensional molecular data, such as RNA sequencing (RNA-seq) and whole-exome sequencing (WES), will likely increase the complexity of clinical interpretation. We compared the recommendations by two molecular tumor boards (MTBs) that independently interpreted the same high-dimensional molecular profiles. Methods: Patients with advanced solid tumors, no available standard therapy, an ECOG performance status of 0-1, and available fresh-frozen tissue underwent WES of tumor tissue and normal blood as well as RNA-seq of tumor tissue within the MASTER (Molecularly Aided Stratification for Tumor Eradication Research) precision oncology program of the German Cancer Consortium (DKTK). Data from 46 patients (WES and RNA-seq, n = 41; WES alone, n = 5) were independently discussed by two MTBs. Treatment recommendations were compared with regard to levels of evidence, therapeutic baskets, and types of biomarkers. Results: A total of 51,610 aberrations (median, 393 per patient) were considered for clinical interpretation (34,314 mutations/single-nucleotide variants, 7,115 mRNA expression changes, 6,144 DNA copy number variations, 4,037 gene fusions). 110 and 132 treatment options were identified by the two MTBs, respectively, with an overall agreement rate of 44.1%. The highest agreement rates were identified for treatment options based on clinical levels of evidence (Level 1, 60%; Level 2, 49.6%) and for poly(ADP-ribose)-polymerase inhibition (57.1%). The lowest agreement rates were identified when MTBs opted for traditional chemotherapy (0%), combination therapies (6.9%), therapies based on preclinical levels of evidence (Level 3, 35.9%; Level 4, 32%), and MAPK inhibition (35%). Similar agreement rates, ranging from 39% (gene fusions) to 54% (loss of heterozygosity), were observed for different types of biomarkers. Conclusions: Reproducible, evidence-based annotation of high-dimensional molecular data is feasible. Our experience provides a basis for ongoing harmonization and standardization efforts within the MTBs of the DKTK.
Ultrasensitive multiplex detection of structural rearrangements in ALK, RET, ROS1 and PD-L1 using a comprehensive next-generation sequencing assay.
Authors: Ngeow Kao Chin , Choudhury Yukti , Tan Kian-Hin , Pek Michelle , Ng Kim-Tien , Lim Soon Thye , Lim Jing Quan , Huang Dachuan , Ong Choon Kiat , Tan Min-Han
Published: May 2020

3572

Background: Oncogenic structural rearrangements (SR) in ALK, RET and ROS1 are well-described in lung cancer, and confer sensitivity to targeted therapy. SR disrupting the 3’UTR of PD-L1 gene have been reported in multiple cancer types and can potentially predict response to checkpoint immunotherapy. An amplicon-based next-generation sequencing (NGS) platform technology (AmpliMARK), previously optimized for detection of single nucleotide variations (SNVs), microsatellite instability and viral DNA, was extended to the multiplex detection of SR in ALK, RET, ROS1 and PD-L1 in cell-free DNA (cfDNA) and tumor tissue DNA. Methods: A hybrid primer-extension and adapter-ligation based method allowing detection of SR in a fusion-partner agnostic manner was utilized for multiplex target capture of genomic regions of ALK, RET, ROS1 and PD-L1 SR. Analytical validation was performed using admixtures of fragmented genomic DNA from an ALK SR-positive cell line, commercial standards containing RET and ROS1 SR, and synthetic PD-L1 SR gene constructs. Clinical performance was assessed in cfDNA samples from lung cancer patients and tumor tissue DNA samples from natural killer(NK)/T-cell lymphoma patients. Results: Detection of SR could be achieved to an allele frequency detection limit of 0.5% with sensitivity of 89.5% and specificity of 100% in admixture samples mimicking cfDNA. In an unselected series of 374 lung cancer cases, actionable SR for ALK, RET and ROS1 were detected in cfDNA of 9 samples, for an overall detection rate of 2.4%, and 1.8% (3 out of 168) when restricted to treatment-naive lung cancer cases only. In 29 NK/T-cell lymphoma tumor tissue samples, 9 samples were positive for PD-L1 SR, which were orthogonally confirmed by whole-genome sequencing, targeted sequencing or Sanger sequencing for a concordance rate of 100% across all samples. For 1 NK/T-cell lymphoma tumor tissue sample where matched plasma was available, the same PD-L1 SR was also detected in cfDNA. Conclusions: We have demonstrated and validated a comprehensive amplicon-based NGS assay for ultrasensitive multiplex detection of structural rearrangements in ALK, RET, ROS1 and PD-L1 across both cfDNA and tumor tissue DNA in analytical and clinical contexts. Ongoing studies will further evaluate the performance and utility of this assay across a larger number of clinical samples for the detection of these SR as well as additional cancer-associated SR involving NTRK1/2/3, FGFR2/3 and TMPRSS2.