73 Background: For patients with RAS/RAF wild type metastatic colorectal cancer (mCRC), systemic therapy usually includes fluoropyrimidine-based chemo + anti-VEGF or anti-EGFR targeted therapy. The PARADIGM trial established tumor sidedness as an important predictive biomarker for patients with RAS/RAF wild-type disease; in right-sided tumors, there was no survival difference between first-line chemo + anti-EGFR vs. chemo + anti-VEGF. However, there is no comparative data to guide second-line treatment decisions in this population. We aim to compare the effectiveness of second-line chemo + anti-EGFR versus chemo + anti-VEGF therapy for patients with RAS/RAF wild-type, right-sided mCRC who received first-line chemo + anti-VEGF. Methods: We used the nationwide Flatiron Health electronic health record-derived database, comprising de-identified patient-level structured and unstructured data obtained from ~280 cancer clinics and curated via technology-enabled abstraction. Patients ≥18 years old with RAS/RAF wild-type, right-sided mCRC who received first-line therapy consisting of chemotherapy ((FOLFIRI or FOLFOX or CAPEOX) + anti-VEGF and initiated second-line chemotherapy with either anti-EGFR or anti-VEGF targeted therapy between January 2013-May 2024 were included. Multiple imputation with chained equations imputed missing values for sidedness, RAS/RAF status, and propensity score covariates (age, gender, year of diagnosis, synchronous/metachronous disease, MMR/MSI status, ECOG score, CEA level, and first-line therapy duration). Cox proportional hazards modeling with stabilized inverse probability of treatment weighting (IPTW) assessed the association of anti-EGFR vs anti-VEGF treatment with overall survival. Results: 4,444 patients received appropriate first-line treatment and received chemo + anti-EGFR or chemo + anti-VEGF in the second line. Across 25 imputations, an average of 444 patients met inclusion criteria: 175 patients received chemo + anti-EGFR and 269 patients received chemo + anti-VEGF. Following IPTW, baseline characteristics were balanced between treatment groups. Patients who received chemotherapy + anti-EGFR had a 24% increased hazard of death when compared with patients who received chemotherapy + anti-VEGF, though this was not statistically significant (HR 1.24, 95% CI 0.96 – 1.61, p=0.097). Conclusions: Among patients with RAS/RAF wild-type, right-sided mCRC who received first-line chemotherapy + anti-VEGF, there is some evidence to support continuing anti-VEGF therapy vs. switching to anti-EGFR therapy in the second line, though the result was not statistically significant. Future studies should explore predictive biomarkers for EGFR vs VEGF-directed treatment for this population to determine the patients most likely to benefit from anti-EGFR therapy during their disease course.

Supplemental tables: (1) Subject characteristics. (2) Cytogenetic profiles and high-risk features. (3) Prior treatment exposures and refractoriness. (4) CAR T cell product characteristics. (5) Products that did not meet target dose. (6) Adverse events of grade 3-4. (7) Cytokine release syndrome and ICANS. (8). Maintenance therapy. Supplemental Figures: (1) Study schematic and subject disposition, (2) Correlates of manufacturing success, (3) Hematopoietic recovery, (4) Post-infusion T cell phenotypes, (5) Correlates of in vivo expansion and manufacturing success, (6) Late post-infusion CAR T cell re-expansion, (7) Soluble BCMA, (8) Late-onset clinical responses, (9) MM cell BCMA expression, (10) Pre- and post-treatment Sox2-specific T cell responses in CART-BCMA monotherapy patients, (11) Pre- and post-treatment Sox2-specific T cell responses in CART-BCMA + huCART19 combination therapy patients, (12) Sustained post-treatment SOX2-specific T-cell responses.

Patients with advanced pancreatic ductal adenocarcinoma (PDAC) have a median survival of less than a year, highlighting the urgent need for treatment advancements. We report on a phase 1 clinical trial assessing the safety and feasibility of intravenous and local administration of anti-mesothelin CAR T cells in patients with advanced PDAC. While therapy is well tolerated, it demonstrates limited clinical efficacy. Analyses of patient samples provide insights into mechanisms of treatment resistance. Single-cell genomic approaches reveal that post-infusion CAR T cells express exhaustion signatures, including previously identified transcription factors ID3 and SOX4, and display enrichment for a GZMK+ phenotype. Single knockout of ID3 or SOX4 enhances efficacy in xenograft models, though with donor-dependent variability. However, single-knockout cells eventually fail. Conversely, ID3 and SOX4 double-knockout CAR T cells exhibit prolonged relapse-free survival, demonstrating a sustained therapeutic effect and a potential avenue for engineering more potent CAR T cells in PDAC. This study was registered at ClinicalTrials.gov (NCT03323944).

Supplemental Table 1: Reasons for MajesTEC-1 ineligibility

Lisocabtagene maraleucel (liso-cel) is an autologous CD19-directed CAR T cell therapy approved for the treatment of relapsed/refractory large B-cell lymphoma (LBCL). We present a multicenter retrospective study evaluating safety, efficacy, and resource utilization of liso-cel in the standard-of-care setting. Patients received commercial liso-cel at 7 US medical centers and patient selection, toxicity management, and disease assessment followed institutional practices. Among 101 infused patients, the median age was 71 years (35% ≥75 years), 68% had a Charlson Comorbidity score ≥3, and 10% had secondary CNS involvement. Median number of prior therapies was 3, and due to comorbidities, 33% would have been ineligible for the TRANSCEND study. Bridging therapy was used in 60% (43% received polatuzumab-based treatment). Any Grade CRS occurred in 49% (3% Grade ≥3) with any Grade ICANS in 26% (10% Grade ≥3). The overall response rate (ORR) to bridging therapy was 45% with 18% achieving a complete response (CR). Following liso-cel infusion, the day 90 ORR was 66% (60% CR), and with a median follow-up of 15.5 months, 12-month progression-free survival (PFS) and overall survival were 55% and 68%, respectively. A normal lactate dehydrogenase pre-lymphodepletion was associated with improved PFS and OS. These analyses confirm similar efficacy and safety of commercial liso-cel compared to pivotal trial results. Notably, these outcomes were achieved in patients predominantly of advanced age and with significant comorbidities. Results also likely reflect advancements in patient selection, toxicity management, and the use of novel bridging strategies.

Long-term risks of gene therapy are not fully understood. In this study, we evaluated safety outcomes in 783 patients over more than 2,200 total patient-years of observation from 38 T cell therapy trials. The trials employed integrating gammaretroviral or lentiviral vectors to deliver engineered receptors to target HIV-1 infection or cancer. Eighteen patients (2.3

e15591 Background: The PARADIGM trial showed an OS benefit to 1L doublet chemo+anti-EGFR vs doublet+anti-VEGF treatment (tx) in patients with RAS wild-type (WT) metastatic colorectal cancer (mCRC) with left-sided primary tumors and some evidence of harm in those with right-sided tumors. Subsequent ctDNA analysis found that a “hyperselection” (HS) biomarker was predictive of anti-EGFR tx benefit, including within subgroups with left- and right-sided tumors. We evaluated the predictive value of a tissue HS biomarker for anti-EGFR benefit in a real-world setting. Methods: We performed a retrospective cohort study using the nationwide Flatiron Health-Foundation Medicine (FMI) mCRC clinico-genomic database. The de-identified data originated from ~280 US cancer clinics. Adults with RAS/RAF WT, MMRp/MSS mCRC who received 1L tx including chemo+ anti-VEGF or anti-EGFR tx from 1/2012 - 12/2024 and who underwent FMI comprehensive genomic profiling on tissue collected pre-1L tx were eligible. Patients without alterations in the HS panel ( ERBB2 amp or activating alterations, MET amp, NTRK/ROS1/RET rearrangement, PIK3CA exon 20/ PTEN inactivating/ AKT1/EGFR ECD alterations) were classified as HS-; those with any alteration were HS+. Multiple imputation was performed for missing values of pre-specified covariates. Multivariable Cox proportional hazards modeling of the association of anti-EGFR vs anti-VEGF tx with OS that included imbalanced covariates and the interaction between tx and HS status was performed in the overall population. Separate models were performed within subgroups defined by tumor sidedness and HS status with adjustment for imbalanced covariates, when possible. Results: 1,332 patients met inclusion criteria, of which 1,155 were HS- and 177 were HS+ (Table). In the multivariable model, the interaction between HS status and tx was not significant (p = 0.51). In subgroup models, there was an 5% reduction in hazard of death for anti-EGFR vs anti-VEGF tx among HS- patients (HR 0.95, 95% CI 0.80 – 1.15) and an 8% increased hazard of death among HS+ patients (HR 1.08, 95% CI 0.69 – 1.69); neither result was statistically significant. Within tumor sidedness subgroups, similar patterns were observed (Table). Conclusions: While there may be some evidence of a differential tx effect by HS status, power to demonstrate predictive effect was limited by the low frequency of alterations in the panel. Among patients with RAS/RAF WT, MMRp/MSS mCRC, tissue HS was not predictive of 1L anti-EGFR tx benefit. Subgroup multivariable models. Hyperselection Negative (n= 1,155) Hyperselection Positive (n=177) Population Anti-VEGF Anti-EGFR HR for OS (95% CI) Anti-VEGF Anti-EGFR HR for OS (95% CI) Overall 870 285 0.95 (0.80-1.15) 135 42 1.08 (0.69 – 1.69) Left-Sided 703 255 0.88 (0.72 – 1.07) 95 34 1.15 (0.68 – 1.97) Right-Sided 167 30 0.94 (0.54 – 1.63) 40 8 1.78 (0.53 – 5.92)

This comparative effectiveness study investigates anti–vascular endothelial growth factor vs anti–epidermal growth factor receptor therapy in second-line treatment among patients with RAS/RAF wild-type, right-sided cancer.

Anti-CD19 CAR (CAR19) T-cell therapy has revolutionized the treatment for relapsed and refractory B-cell malignancies, showing long-term efficacy and potential cures. However, while many patients, including those with Chronic Lymphocytic Leukemia (CLL), exhibit initial signs of response, nearly two-thirds will eventually relapse. This highlights the critical need to understand treatment failure mechanisms and to identify targets to improve therapies. Our team previously reported an exceptional response in a CLL patient treated with CAR19 T-cells, which coincided with significant expansion of a CD8+ CAR T-cell clone harboring a disruption of the TET2 gene from an integration event of the lentiviral construct. Building on this finding, we conducted a large study analyzing lentiviral vector integration sites (LVIS) in 40 CAR19-treated Acute Lymphoblastic Leukemia (ALL) and CLL patients. Consistent with other studies, our data revealed a correlation between LVIS and therapeutic outcomes. To test whether these vector-mediated gene disruptions influence CAR T-cell growth kinetics and anti-tumor efficacy, we designed a CRISPR library targeting 180 LVIS-associated genes using 4-8 guide RNAs (gRNAs) per gene, including positive and negative controls. Our custom vector was engineered to drive the expression of both the anti-CD19 CAR and gRNAs under the promoters EF1a and human U6, respectively. To generate these modified CAR T-cells, we first transduced healthy donor T-cells at low multiplicity of infection, ensuring single-gene edits per cell, purified for CAR positivity three days post-transduction, and expanded cells for additional 10 days. The impact of each gene edit on CAR T-cell fitness was assessed in our well-established in vitro repeat tumor challenge and an in vivo xenogeneic model using the aggressive leukemia cell line NALM6. In both scenarios, we identified several genes with increased frequency (Log2FC ≥ 0.25, FDR ≤ 0.1) upon Robust Rank Aggregation (RRA) analysis using the MAGeCKFlute package in R, suggesting a potential fitness advantage. To validate the top candidates from our screen, we utilized Cas9-ribonucleoprotein (RNP) complexes with 2 independent gRNAs compared to AAVS1 (safe-targeting) and wild type controls. The targeted disruptions of the genes PCNX1 and PDCD10, among others, exhibited superior proliferative capacity upon our in vitro assay compared to controls. Ongoing studies include in vivo validation, phenotypic analyses of all candidates through spectral flow cytometry, and RNA sequencing to elucidate the biological roles of these genes in T-cell function. This preclinical package has the potential to advance into a Phase 1 clinical trial, where our optimized CAR T-cell product will be tested in CLL and potentially other refractory/relapsed B-cell malignancies. Beatriz Coutinho de Oliveira, Saaurav Bari, Jessica Kerr, Victoria Putzbach, Yee Peng Phoon, Joseph A. Fraietta, Wei-Ting Hwang, Frederic D. Bushman, Jan Joseph Melenhorst. Validation of a functional genomics screen based on lentiviral integration site analysis reveals single-gene disruptions enhancing CAR19 T-cell effectiveness in preclinical models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3807.

BACKGROUND:Chimeric antigen receptor (CAR) T cells targeting CD19 have transformed the treatment of B-cell cancers, but many patients do not have long-term remission. We designed an anti-CD19 enhanced (armored) CAR T-cell product (huCART19-IL18) that secretes interleukin-18 to enhance antitumor activity. METHODS:In this study, we assessed the safety, feasibility, and preliminary efficacy of huCART19-IL18 in patients with relapsed or refractory lymphoma after previous anti-CD19 CAR T-cell therapy. Using a 3-day manufacturing process, we administered huCART19-IL18-positive cells in doses ranging from 3×106 to 3×108. RESULTS:A total of 21 patients received huCART19-IL18. Cytokine release syndrome occurred in 62% of the patients (47% with grade 1 or 2), and immune effector-cell-associated neurotoxicity syndrome occurred in 14% (all grade 1 or 2). No unexpected adverse events were observed. Robust CAR T-cell expansion was detected across all dose levels. At 3 months after infusion, a complete or partial response was seen in 81% of the patients (90% confidence interval [CI], 62 to 93) and a complete response in 52% (90% CI, 33 to 71). With a median follow-up of 17.5 months (range, 3 to 34), the median duration of response was 9.6 months (90% CI, 5.5 to not reached). CONCLUSIONS:In this small study, huCART19-IL18 had a safety profile consistent with other CAR T-cell treatments and showed promising efficacy at low cell doses in patients with lymphoma after the failure of previous anti-CD19 CAR T-cell therapy. (ClinicalTrials.gov number, NCT04684563.).

Primary central nervous system lymphoma (PCNSL) is a rare form of aggressive non-Hodgkin lymphoma. Given its infrequency, there are few randomized trials to guide induction and consolidation strategies, with no consensus on optimal treatment. Most centers will offer high-dose methotrexate-based induction chemotherapy followed by either autologous stem cell transplant, whole-brain radiation, or prolonged chemotherapy. The preferred strategy at our institution has been 6 doses of methotrexate, temozolomide, and rituximab (MTR) induction with methotrexate on day 15 until complete response for induction followed by 6 monthly cycles of MTR. We conducted a retrospective analysis of patients diagnosed with PCNSL at the University of Pennsylvania from 1 April 2008 to 1 October 2024, identifying 153 patients who received this regimen. With a follow-up of 63 months, the median overall survival (OS) in the entire cohort was 65 months, with a median relapse-free survival (RFS) of 36 months. In the cohort of patients who were able to complete 6 months of MTR induction and proceed with MTR consolidation, median OS and RFS were 143 and 122 months, respectively. Although 13% of patients discontinued therapy because of toxicity, there was no treatment-related mortality. These results indicate that prolonged MTR is a safe treatment option and an alternative to intensified consolidation strategies. Further randomized studies are necessary to determine the optimal treatment strategy in newly diagnosed PCNSL.

102 Background: Outcomes in patients with rGBM are poor, with historical median overall survival (OS) of 6-9 months. Here we report the results from the dose exploration phase of a phase 1 trial investigating ICV-delivered, bivalent CAR T-cells targeting EGFR epitope 806 and IL13Rα2 in rGBM. Methods: Patients with EGFR-amplified GBM that was recurrent/progressive following front-line radiotherapy were enrolled using a 3+3 design (dose levels: 5.0 x 10 6 , 1.0 x 10 7 , and 2.5 x 10 7 cells). Patients underwent surgery for (1) maximal safe resection and confirmation of viable tumor and (2) Ommaya reservoir placement. Post-operatively, patients received a single ICV dose of CART-EGFR-IL13Rα2 cells without lymphodepleting chemotherapy. Primary endpoints included dose-limiting toxicity (DLT) and determination of the maximum tolerated dose (MTD). Secondary endpoints included objective radiographic response, progression-free survival (PFS), and OS. Serial CSF samples were analyzed for CAR T-cell pharmacokinetics and single-cell RNA sequencing (scRNAseq). Results: Eighteen patientsreceived CART-EGFR-IL13Rα2 cells (n=6 per dose level). Median age was 57, 15 (83%) were male, 13 (72%) had MGMT unmethylated tumors, and 7 (39%) had >1 prior relapse. One DLT (grade 3 lethargy/fatigue) was observed at the MTD (2.5 x 10 7 cells). Acute neurotoxicity related to CAR T-cells occurred in all patients. Using immune effector cell-associated neurotoxicity syndrome (ICANS) grading, 10 of 18 patients (56%) experienced grade 3 neurotoxicity; none had grade 4-5 neurotoxicity. Using tumor-inflammation associated neurotoxicity (TIAN) grading, 2 of 18 patients (11%) had grade 3 and 1 patient (6%) had grade 4 neurotoxicity. Grade 1-2 fever occurred in all patients. Eleven of 13 patients (85%) with measurable disease at time of CAR T-cell infusion experienced tumor shrinkage, ranging from 1-62% reductions (median 35%, IQR 12 – 39%) in target lesions and with one confirmed partial response by modified RANO criteria. PFS and OS continue to mature and will be presented. CAR T-cell expansion in CSF was robust with a dose-response relationship observed. The CAR transgene remained detectable in CSF and blood 12 months post-CART infusion in a patient experiencing durable stable disease lasting for 17 months (ongoing at data cut-off). In patients undergoing repeat resection following treatment, CART-EGFR-IL13Rα2 cell infusion markedly increased the number of tumor-infiltrating lymphocytes. scRNAseq in post-treatment CSF revealed higher cytotoxicity and exhaustion scores in CD8+ CAR T-cells as compared to the infusion product, indicative of target cell engagement. Conclusions: ICV delivery of CART-EGFR-IL13Rα2 is feasible and appears safe. CART-EGFR-IL13Rα2 cells are bioactive and exhibit an encouraging early efficacy signal in rGBM. Clinical trial information: NCT05168423 .

Glioblastoma (GBM) is the most common primary brain cancer in adults and carries a median overall survival (OS) of 12-15 months. Effective therapy for recurrent GBM (rGBM) following frontline chemoradiation is a major unmet medical need. Here we report the dose escalation and exploration phases of a phase 1 trial investigating intracerebroventricular delivery of bivalent chimeric antigen receptor (CAR) T cells targeting epidermal growth factor receptor (EGFR) epitope 806 and interleukin-13 receptor alpha 2 (IL-13Rα2), or CART-EGFR-IL13Rα2 cells, in patients with EGFR-amplified rGBM. Primary endpoints included dose-limiting toxicity, determination of the maximum tolerated dose and recommended dose for expansion, and occurrence of adverse events. Secondary endpoints included objective radiographic response, duration of response, progression-free survival and OS. A total of 18 patients received CART-EGFR-IL13Rα2 cells. The maximum tolerated dose was determined to be 2.5 × 107 cells. Of the 18 patients, 10 (56%) experienced grade 3 neurotoxicity; none had grade 4-5 neurotoxicity. Of 13 patients, 8 (62%) with measurable disease at the time of CAR T cell infusion experienced tumor regression, with one confirmed partial response by Modified Response Assessment in Neuro-Oncology criteria (objective radiographic response, 8%; 90% confidence interval, 0-32%) and one patient with ongoing durable stable disease lasting over 16 months. Median progression-free survival was 1.9 months (90% confidence interval, 1.1-3.4 months), and median OS was not yet reached at the time of data cut-off (median follow-up time, 8.1 months). These findings indicate that intracerebroventricular delivery of bivalent CART-EGFR-IL13Rα2 is feasible and appears safe. CART-EGFR-IL13Rα2 cells are bioactive and exhibit a signal of antitumor effect in rGBM. ClinicalTrials.gov registration: NCT05168423 .

Background: The KEYNOTE-048 study established the checkpoint inhibitor (CPI) pembrolizumab, with/without chemotherapy, as frontline treatment for recurrent/metastatic (R/M) head and neck squamous cell carcinoma (HNSCC). However, pembrolizumab monotherapy has limited efficacy in PD-L1–negative disease. Clinical practice patterns regarding PD-L1 combined positive score (CPS) testing and PD-L1–guided treatment selection remain unknown. Patients and Methods: This retrospective analysis included patients who initiated treatment for R/M HNSCC from 2011 to 2023 in a nationwide electronic health record–derived deidentified database. Frontline therapy was categorized as CPI monotherapy, CPI with chemotherapy, or chemotherapy ± cetuximab without CPI. A subset of patients treated in 2019 and beyond (2019+ cohort) were analyzed to investigate PD-L1 testing rates, treatment patterns following FDA approval of pembrolizumab, and the proportion receiving “off-label” CPI monotherapy (single-agent use in patients with metastatic HNSCC and negative/unknown PD-L1 status). Factors associated with “off-label” use were identified using multivariable logistic regression. Results: The total cohort included 7,657 patients with a median age of 65 years (IQR, 58–72); 67% were White, 78% had a history of smoking, 66% had an ECOG performance status (PS) of 0–1, and 31% were HPV-positive. The 2019+ subset included 3,395 patients, of whom nearly half (47%) did not have a known PD-L1 CPS prior to systemic treatment initiation. The most common frontline treatment in the total cohort was CPI monotherapy (43%). CPI monotherapy use was even higher in patients aged ≥75 years (54%) and those with ECOG PS ≥2 (52%). Among the 2019+ subgroup with PD-L1 CPS negative/unknown tumors (n=1,926), 536 (28%) received CPI monotherapy “off-label.” Factors associated with “off-label” use on multivariable regression included age ≥75 years (odds ratio [OR], 1.4), community practice setting (OR, 1.5), and earlier year of treatment (OR, 1.3 per year) (all P <.05). Conclusions: Most US patients with R/M HNSCC are now receiving CPI-based therapy in the frontline setting; however, PD-L1 testing remains underutilized. “Off-label” use of CPI monotherapy in PD-L1–negative/unknown HNSCC is common, particularly among elderly patients.

Abstract Purpose: Addition of ataxia telangiectasia and Rad3-related kinase inhibitors (ATRi) to poly-ADP ribose polymerase inhibitors (PARPi) overcomes PARPi-resistance in high grade serous ovarian cancer (HGSOC) cell and mouse models. We present the results of an investigator-initiated study of combination PARPi(olaparib) and ATRi(ceralasertib) in patients with acquired PARPi-resistant HGSOC. Patients and Methods: Eligible patients had recurrent, platinum-sensitive BRCA1/2 mutated or homologous recombination (HR) deficient HGSOC and clinically benefited from PARPi (response by imaging/CA-125 or duration of maintenance therapy; >12 months 1st-line or >6 months ≥2nd-line) before progression. No intervening chemotherapy was permitted. Patients received olaparib 300mg twice daily and ceralasertib 160mg daily on days 1-7 of a 28-day cycle. Primary objectives were safety and objective response rate (ORR). Results: Thirteen patients enrolled were evaluable for safety and 12 for efficacy. 62%(n=8) had germline BRCA1/2 mutations, 23% (n=3) somatic BRCA1/2 mutations, and 15%(n=2) HR-deficient tumors. Prior PARPi indication was treatment for recurrence (54%, n=7), 2nd line-maintenance (38%, n=5), and frontline treatment with carboplatin/paclitaxel (8%, n=1). There were 6 partial responses yielding an ORR of 50% (95% CI:0.15, 0.72). Median treatment duration was 8 cycles (range 4-23+). Grade(G) 3/4 toxicities were 38%(n=5); 15%(n=2) G3 anemia, 23%(n=3) G3 thrombocytopenia, 8% (n=1) G4 neutropenia. Four patients required dose-reductions. No patient discontinued treatment due to toxicity. Conclusion: Combination olaparib and ceralasertib is tolerable and shows activity in HR-deficient platinum-sensitive recurrent HGSOC that benefited and then progressed with PARPi as the penultimate regimen. These data suggest that ceralasertib re-sensitizes PARPi resistant HGSOCs to olaparib, warranting further investigation.

PURPOSE Less than half of the patients with newly diagnosed metastatic non-small cell lung cancer (NSCLC) undergo comprehensive molecular testing. We designed an electronic medical record (EMR)-based "nudge intervention" to prompt plasma-based molecular testing at the time of initial medical oncology consultation. METHODS A nonrandomized prospective trial was conducted at the University of Pennsylvania's academic practice and two affiliated community practices. Molecular genotyping was performed by tissue- and/or plasma-based next generation sequencing methods. Comprehensive testing was defined as testing for EGFR, ALK, BRAF, ROS1, MET, RET, KRAS, and NTRK. Guideline-concordant treatment was defined as the use of the appropriate first-line (1L) therapy as per the National Comprehensive Cancer Network (NCCN) guidelines. Proportion of patients with comprehensive molecular genotyping results available at any time, molecular results available before 1L therapy, and guideline-concordant 1L treatment were compared between the preintervention and postintervention cohorts using Fisher's exact test or Pearson's chi-squared test. RESULTS Five hundred and thirty-three patients were included, 376 in the preintervention cohort and 157 in the postintervention cohort. After implementation of the EMR-based nudge, a higher proportion of patients underwent comprehensive molecular testing in the postintervention versus the preintervention cohort (100% v 88%, P = <.001), had results of comprehensive molecular testing available before initiating 1L treatment (97.3% v 91.6%, P = .026), and received NCCN guideline-concordant care (89.8% v 78.2%, P = .035). CONCLUSION Across three practice sites in a large health system, implementation of a provider team-focused EMR-based nudge intervention was feasible, and led to a higher number of patients with NSCLC undergoing comprehensive molecular genotyping. These findings demonstrate that behavioral nudges can promote molecular testing and should be studied further as a tool to improve guideline-concordant care in both community and academic sites.

The use of lentiviral vectors in cell and gene therapy is steadily increasing, both in commercial and investigational therapies. Although existing data increasingly support the usefulness and safety of clinical-grade lentiviral vectors used in cell manufacturing, comprehensive studies specifically addressing their long-term stability are currently lacking. This is significant considering the high cost of producing and testing GMP-grade vectors, the limited number of production facilities, and lengthy queue for production slots. Therefore, an extended shelf life is a critical attribute to justify the investment in large vector lots for investigational cell therapies. This study offers a thorough examination of essential stability attributes, including vector titer, transduction efficiency, and potency for a series of clinical-grade vector lots, each assessed at a minimum of 36 months following their date of manufacture. The 13 vector lots included in this study were used for cell product manufacturing in 16 different clinical trials, and at the time of the analysis had a maximum storage time at -80 degrees C of up to 8 years. The results emphasize the long-term durability and efficacy of GMP-grade lentiviral vectors for use in ex vivo cell therapy manufacturing.

We previously showed that chimeric antigen receptor (CAR) T-cell therapy targeting epidermal growth factor receptor variant III (EGFRvIII) produces upregulation of programmed death-ligand 1 (PD-L1) in the tumor microenvironment (TME). Here we conducted a phase 1 trial (NCT03726515) of CAR T-EGFRvIII cells administered concomitantly with the anti-PD1 (aPD1) monoclonal antibody pembrolizumab in patients with newly diagnosed, EGFRvIII+ glioblastoma (GBM) (n = 7). The primary outcome was safety, and no dose-limiting toxicity was observed. Secondary outcomes included median progression-free survival (5.2 months; 90% confidence interval (CI), 2.9-6.0 months) and median overall survival (11.8 months; 90% CI, 9.2-14.2 months). In exploratory analyses, comparison of the TME in tumors harvested before versus after CAR + aPD1 administration demonstrated substantial evolution of the infiltrating myeloid and T cells, with more exhausted, regulatory, and interferon (IFN)-stimulated T cells at relapse. Our study suggests that the combination of CAR T cells and PD-1 inhibition in GBM is safe and biologically active but, given the lack of efficacy, also indicates a need to consider alternative strategies.

Introduction: Clinical trials evaluating chimeric antigen receptor T-cell therapy (CAR T) commonly report time-to-event (TTE) endpoints. However, definitions are not necessarily comparable across studies and variability can lead to misinterpretation of results or inappropriate comparisons across products and studies. Amid the rapidly increasing number of published CAR T trials-many of which were used for regulatory approval-this study aims to summarize the variation in the use and reporting of TTE endpoints in CAR T trials. Methods: We include CAR T trials published January 2008-January 2023 on PubMed that reported at least one of these TTE endpoints: overall survival (OS), progression-free survival (PFS), duration of response/remission (DOR), disease-free survival, event-free survival (EFS), relapse-free survival (RFS), time to relapse, time to progression, or time to treatment failure. We abstracted and summarized endpoint definitions, including the time origin, events, competing events, and censoring. We assessed the completeness of endpoint reporting, overall and by subgroups such as study phase, publication year, and the journal's impact factor. Results: We included 116 publications in the analysis. The most frequently reported TTEs were OS (83%,), PFS (56%), DOR (55%), and EFS (23%). Complete reporting of endpoints was poor overall: 32%, 24%, 25%, and 56% for OS, PFS, DOR, and EFS respectively. Complete reporting was lower in articles published before 2018, in lower impact factor journals, and in phase I trials. There was also a large variability in TTE definitions among those reported. For example, among 64 studies reporting DOR, 48% used the date of response as the time origin while 20% used the date of infusion, and 31% did not report a time origin. Conclusion: There is substantial heterogeneity and incompleteness of TTE endpoint definitions in CAR T trials reported in the existing literature that could impact the interpretation of the study results. Improving and harmonizing reporting is required to ensure valid assessment of clinical benefit and cross-trial comparison. Complete reporting should include all the following components: time origin, event(s) of interest, competing event(s) if any, and censoring. Recommended definitions for the common TTEs and the rationales will be presented at the meeting.