Explore the vast range of titles available.

Axicabtagene ciloleucel is an autologous anti-CD19 chimeric antigen receptor (CAR) T-cell therapy. In the previous analysis of the ZUMA-1 registrational study, with a median follow-up of 15·4 months (IQR 13·7–17·3), 89 (82%) of 108 assessable patients with refractory large B-cell lymphoma treated with axicabtagene ciloleucel achieved an objective response, and complete responses were noted in 63 (58%) patients. Here we report long-term activity and safety outcomes of the ZUMA-1 study. ZUMA-1 is a single-arm, multicentre, registrational trial at 22 sites in the USA and Israel. Eligible patients were aged 18 years or older, and had histologically confirmed large B-cell lymphoma—including diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, and transformed follicular lymphoma—according to the 2008 WHO Classification of Tumors of Hematopoietic and Lymphoid Tissue; refractory disease or relapsed after autologous stem-cell transplantation; an Eastern Cooperative Oncology Group performance status of 0 or 1; and had previously received an anti-CD20 monoclonal antibody containing-regimen and an anthracycline-containing chemotherapy. Participants received one dose of axicabtagene ciloleucel on day 0 at a target dose of 2 × 106 CAR T cells per kg of bodyweight after conditioning chemotherapy with intravenous fludarabine (30 mg/m2 body-surface area) and cyclophosphamide (500 mg/m2 body-surface area) on days −5, −4, and −3. The primary endpoints were safety for phase 1 and the proportion of patients achieving an objective response for phase 2, and key secondary endpoints were overall survival, progression-free survival, and duration of response. Pre-planned activity and safety analyses were done per protocol. ZUMA-1 is registered with ClinicalTrials.gov, number NCT02348216. Although the registrational cohorts are closed, the trial remains open, and recruitment to extension cohorts with alternative endpoints is underway. Between May 19, 2015, and Sept 15, 2016, 119 patients were enrolled and 108 received axicabtagene ciloleucel across phases 1 and 2. As of the cutoff date of Aug 11, 2018, 101 patients assessable for activity in phase 2 were followed up for a median of 27·1 months (IQR 25·7–28·8), 84 (83%) had an objective response, and 59 (58%) had a complete response. The median duration of response was 11·1 months (4·2–not estimable). The median overall survival was not reached (12·8–not estimable), and the median progression-free survival was 5·9 months (95% CI 3·3–15·0). 52 (48%) of 108 patients assessable for safety in phases 1 and 2 had grade 3 or worse serious adverse events. Grade 3 or worse cytokine release syndrome occurred in 12 (11%) patients, and grade 3 or worse neurological events in 35 (32%). Since the previous analysis at 1 year, additional serious adverse events were reported in four patients (grade 3 mental status changes, grade 4 myelodysplastic syndrome, grade 3 lung infection, and two episodes of grade 3 bacteraemia), none of which were judged to be treatment related. Two treatment-related deaths (due to haemophagocytic lymphohistiocytosis and cardiac arrest) were previously reported, but no new treatment-related deaths occurred during the additional follow-up. These 2-year follow-up data from ZUMA-1 suggest that axicabtagene ciloleucel can induce durable responses and a median overall survival of greater than 2 years, and has a manageable long-term safety profile in patients with relapsed or refractory large B-cell lymphoma. Kite and the Leukemia & Lymphoma Society Therapy Acceleration Program.

The in vivo persistence of adoptively transferred T cells is predictive of antitumor response. Identifying functional properties of infused T cells that lead to in vivo persistence and tumor eradication has remained elusive. We profiled CD19-specific chimeric antigen receptor (CAR) T cells as the infusion products used to treat large B cell lymphomas using high-throughput single-cell technologies based on time-lapse imaging microscopy in nanowell grids (TIMING), which integrates killing, cytokine secretion, and transcriptional profiling. Our results show that the directional migration of CD19-specific CAR T cells is correlated with multifunctionality. We showed that CD2 on T cells is associated with directional migration and that the interaction between CD2 on T cells and CD58 on lymphoma cells accelerates killing and serial killing. Consistent with this, we observed that elevated CD58 expression on pretreatment tumor samples in patients with relapsed or refractory large B cell lymphomas treated with CD19-specific CAR T cell therapy was associated with complete clinical response and survival. These results highlight the importance of studying dynamic T cell-tumor cell interactions in identifying optimal antitumor responses.

BackgroundChimeric antigen receptor (CAR)-T cells are approved for use in the treatment of hematological malignancies. Axicabtagene ciloleucel (YESCARTA) and brexucabtagene autoleucel (TECARTUS) genetically modified autologous T cells expressing an anti-CD19 scFv based on the FMC63 clone have shown impressive response rates for the treatment of CD19+B cell malignancies, but there remain challenges in monitoring long-term persistence as well as the functional characterization of low-level persisting CAR-T cells in patients. Furthermore, due to CD19-negative driven relapse, having the capability to monitor patients with simultaneous detection of the B cell malignancy and persisting CAR-T cells in patient peripheral blood is important for ensuring timely treatment optionality and understanding relapse.MethodsThis study demonstrates the development and technical validation of a comprehensive liquid biopsy, high-definition single cell assay (HDSCA)-HemeCAR for (1) KTE-X19 CAR-T cell identification and analysis and (2) simultaneously monitoring the CD19-epitope landscape on neoplastic B cells in cryopreserved or fresh peripheral blood. Proprietary anti-CD19 CAR reagents, healthy donor transduced CAR-T cells, and patient samples consisting of malignant B cell fractions from manufacturing were used for assay development.ResultsThe CAR-T assay showed an approximate limit of detection at 1 cell in 3 million with a sensitivity of 91%. Genomic analysis was additionally used to confirm the presence of the CAR transgene. This study additionally reports the successful completion of two B cell assays with multiple CD19 variants (FMC63 and LE-CD19) and a unique fourth channel biomarker (CD20 or CD22). In patient samples, we observed that CD19 isoforms were highly heterogeneous both intrapatient and interpatient.ConclusionsWith the simultaneous detection of the CAR-T cells and the B cell malignancy in patient peripheral blood, the HDSCA-HemeCAR workflow may be considered for risk monitoring and patient management.

A current paradigm in immunology is that the strength of T cell responses is governed by antigen dose, localization, and costimulatory signals. This study investigates the influence of antigen kinetics on CD8 T cell responses in mice. A fixed cumulative antigen dose was administered by different schedules to produce distinct dose-kinetics. Antigenic stimulation increasing exponentially over days was a stronger stimulus for CD8 T cells and antiviral immunity than a single dose or multiple dosing with daily equal doses. The same was observed for dendritic cell vaccination, with regard to T cell and anti-tumor responses, and for T cells stimulated in vitro. In conclusion, stimulation kinetics per se was shown to be a separate parameter of immunogenicity. These findings warrant a revision of current immunization models and have implications for vaccine development and immunotherapy.

BackgroundChimeric Antigen Receptor T cells (CARTs) are a powerful anti-cancer therapy, demonstrating success in hematologic malignancies. The development of targeted lipid nanoparticle-mRNA (tLNP-mRNA) therapeutics has allowed for the generation of CARTs in situ and may resolve several challenges of conventional ex vivo viral-engineered CAR T cell products including scalability, access and mulitplexing. The in situ T cell transfection rate achieved by tLNP-mRNA varies from 4–20% of T cells expressing the protein of interest. As tLNP-mRNA platform efficacy may critically depend on the number, metabolic state, and localization of engineered T cells, we investigated whether cytokines could enhance protein expression.MethodsWe used tLNPs that target CD5, a marker expressed highly on mouse and human T cells. These tLNPs carried the mRNA for the reporter protein mCherry or encoded a fibroblast activated protein (FAP) targeted CAR. Mouse and human T cells were cultured with IL2, IL7, IL15 or activated using αCD3/CD28. CD5-mCherry-tLNPs or CD5-FAPCAR-tLNPs were added and protein expression was detected using flow cytometry. For in vivo studies, C57BL/6 mice were pretreated with IL7, injected with tLNPs and sacrificed 24 hours later. To analyze the T cell transcriptome, CD8+ T cells were isolated from mouse spleens and cultured with IL2, IL7 or IL15 for 48 hours before being sequenced.ResultsWe found that CD5-mCherry-tLNPs induced protein expression on 10% of resting T cells in vivo and ~15% of T cells in vivo. Culturing mouse and human T cells with IL7 significantly improved CD5-mCherry-tLNPs protein expression in vitro. This also occurred in the in vivo setting as pre-treating mice with IL7 elevated both the proportion and total number of mCherry expressing T cells. FAPCAR expression was also increased by combining CD5-FAPCAR-tLNPs with recombinant IL7. Transcriptomic analysis showed IL7 selectively increased pathways associated with protein translation. The significance of these transcriptomic changes was demonstrated by showing that after electroporation with mRNA, T cells cultured in IL7 produced more protein compared to IL2 or IL15.ConclusionsT cells can be engineered in situ using CD5-targeted tLNPs and IL7 increases the protein expression induced by tLNPs. Our data suggests that the upregulation of translation-associated pathways in T cells by IL7 could be exploited to improve the expression of proteins in situ after tLNP administration. This provides a novel paradigm through which a T cell activating cytokine, instead of lymphodepletion, can potentiate in situ CAR T cell therapy.Ethics ApprovalThis study was approved by The University of Pennsylvania’s Ethics Board; approval number 806099.

BackgroundAutologous chimeric antigen receptor (CAR) T cell therapies have revolutionized the treatment of some cancers and are now demonstrating effects in autoimmune disease. With several approved therapies on the market and cure rates approaching 50% in various hematologic malignancies, access to these life-saving therapies is paramount. However, challenges with cell manufacturing, scaling, and the need for inpatient treatment necessitate a truly off-the-shelf solution, especially for non-oncology indications. Utilizing the success of mRNA lipid nanoparticles (LNP) as COVID vaccines, Capstan Therapeutics has developed a novel targeted LNP (tLNP) platform that is purpose-built for specific delivery of therapeutic mRNAs to immune cells through functionalization with a targeting antibody.ResultsCapstan has developed rationally designed proprietary LNPs that exhibit significantly reduced delivery to liver compared to conventional LNPs and effectively deliver to T cells when functionalized with a T cell specific targeting antibody. These Capstan proprietary tLNPs were well tolerated following a single intravenous dose up through 6mg/kg in male Sprague Dawley rats, a highly sensitive species for evaluating LNP toxicity. In vitro screens for the optimization of payload mRNA components, including UTR sequences and codon usage, resulted in a several fold increase in CAR expression and promoted improved tumor cell killing. CD5 and CD8 antibody tLNPs delivered a reporter gene mRNA payload to human cells in vivo in a humanized mouse model at high efficiency and specificity. CD8 tLNPs specifically reprogrammed CD8 T cells effectively with minimal reporter expression in CD4 T cells, whereas CD5 tLNPs reprogrammed both CD8 and CD4 T cell populations. Delivery of an anti-CD19 mRNA CAR construct by both CD5 and CD8 tLNPs into a human PBL-engrafted Nalm6 tumor-bearing mouse model resulted in rapid clearance of the tumor. Repeat doses of tLNPs (BIWx5) at dose levels up through 30 µg/animal were well tolerated and the CAR was expressed on T cells in vivo after single and repeat dosing.ConclusionsCapstan’s tLNPs can specifically target and deliver a therapeutic CAR payload in vivo resulting in functional anti-tumor CAR T cells. This non-viral, redosable approach promises to improve access, efficacy, and safety, owing in part to lack of harsh lymphodepletion conditioning. Due to the versatility of this platform, treatments for various disease categories can be envisioned using different targeting binders to deliver a broad set of payloads to diverse cell populations.Ethics ApprovalThis study complied with all relevant ethical regulations and all animal protocols were approved by the Explora BioLabs (AAALAC-accredited) Institutional Animal Care and Use Committee (IACUC).

This Editorial announces a new section in the Journal of Translational Medicine: Patient-Targeted Molecular Therapies. This section is dedicated to the dissemination of targeted molecular therapies in context of patient-centered outcomes research and evidence-based clinical decisions. The focus on patient-targeted molecular therapies – spanning small molecules and biomolecules alike – stems from the unprecedented growth in this arena. This is consonant with the overall objective of the Journal of Translational Medicine, which seeks out to expand firmly to other vast areas of medicine in the domain of translational science, viewed here as the transaction between translational research and translational effectiveness. As we inaugurate this new section in Journal of Translational Medicine, with its mission described in detail in this Editorial, we invite interested scientists to submit their work for publication.

Influenza viruses are among the most significant human pathogens, responsible for increased seasonal morbidity and mortality particularly in immunodepressed and chronically ill. Conventional vaccination with non-replicative vaccine is currently performed by injection. In the present study, we explore simple spray-dried lipid formulations containing whole inactivated virus or split-subunit vaccine that allow aerosolization and thus, mucosal vaccination of the pulmonary tract. We show that by using biocompatible excipients already approved for human use, one could engineer microparticles that induce substantial local and systemic immunity subsequent to pulmonary administration. Exposure of the bronchial-associated lymphoid tissue (BALT) to vaccine was more effective than parenteral or nasal administration in triggering specific immunity. Co-formulation of a biocompatible surfactant detergent greatly ameliorated the immune profile of microparticles containing a whole inactivated virus vaccine. In addition, mere formulation of a licensed split-subunit vaccine significantly enhanced its immunogenicity. Together, our data underline a simple strategy to convert conventional parenteral vaccination of currently available non-replicative vaccines against influenza virus, into one that is more effective and practical upon respiratory administration.

For the last four years the Journal of Translational Medicine (JTM) has hosted the Section of Tumor Immunology and Biological Cancer Therapy. Under the editorial leadership of Dr. Pedro Romero and with the direct support of the Society for Immunotherapy of Cancer (SITC), this section enriched the communication between basic immunological sciences and the clinical investigation arena in oncology. We are re-launching this Section of JTM, now entitled Immunobiology and Immunotherapy, succeeding Tumor Immunology and Biological Cancer Therapy. While aiming to build on the editorial success and focus of its predecessor, this novel Section will have a broader scope, hosting translational immunology topics pertaining to immunotherapy beyond oncology, including disciplines such as inflammation, autoimmunity, transplantation, metabolic disorders and others. As the vision of this re-launched Section of JTM broadens up to serve a communication need for translational immunologists involved with immunotherapy irrespectively of the therapeutic area, a novel and focused journal entitled Journal for Immunotherapy of Cancer (JITC) has just been initiated, sponsored by the SITC.

BackgroundT-cell based immunotherapies such as CAR-T, bispecific mAb, transgenic T cells and checkpoint blockade have profound efficacy in multiple tumor types but share a common limitation – target antigen (Ag) escape.1 2 One approach to address this limitation has been therapy directed at a ‘parallel’ target (e.g. CD22 after CD19 loss), however, these lineage markers are frequently lost together.3 Here, we describe an alternate, broadly applicable, approach: potentiating fasL/fas-signaling to increase localized bystander killing of Ag-tumor cells and thereby prevent Ag escape.MethodsWe used a CRISPR/Cas9 library to screen for tumor expressed molecules that inhibit or facilitate T-cell killing. We then evaluated one candidate -fas- using murine transgenic T cells, murine and human CAR-T cells, bispecific mAb redirected PBMC, and tumoral RNAseq data from a large CAR-T clinical trial.ResultsGFP-specific (JEDI) CD8 T cells were co-cultured with on-target (GFP+) and bystander (mCherry+) lymphoma cells that had been transfected with a CRISPR/Cas9 library; this screen revealed several tumor-expressed candidate molecules inhibiting or facilitating T-cell killing. Notably, we observed a marked dependence on fas for on-target tumor killing and then, surprisingly, an exquisite dependence on fas for localized bystander tumor killing. (figure 1).Because bystander tumor killing appeared critically fas-dependent, we hypothesized that potentiating fas-signaling might increase bystander killing. An in vitro screen of small molecules that modulate fas-pathway revealed several candidates, including inhibitors of histone deacetylases (HDAC), inhibitors of apoptosis proteins (IAP) and Bcl-2 family members in murine and human systems (figure 2). To validate these candidates, we demonstrated that HDACi increased GFP-specific T cell killing of both on-target and bystander lymphoma cells, in a completely fas-dependent manner (figure 3). Similarly, using a bispecific antibody-based system, we demonstrated increased, fas-dependent, T cell killing of both on-target and bystander human lymphoma cells with inhibitors of IAP and bcl-2 family members (e.g. MCL1).Abstract 90 Figure 1See text for descriptionAbstract 90 Figure 2See text for descriptionAbstract 90 Figure 3See text for descriptionConclusionsT-cell mediated tumor killing can be potentiated with fas pathway modulators. This augmentation improves both fas-dependent Ag+ and Ag-tumor cell death. Further studies of modulating the fas pathway alongside T-cell based immunotherapies are needed as potential treatments to prevent antigen escape and improve patient outcomes.AcknowledgementsWe thank the flow cytometry core facility, microscopy core facility, and the CCMS animal facility at ISMMS.Ethics ApprovalThe studies were approved by The Mount Sinai Institutional Review Board.ReferencesZaretsky J, Garcia-Diaz A, Shin D, et al. Mutations Associated with Acquired Resistance to PD-1 Blockade in Melanoma. N Engl J Med 2016: 375(9); 819–20.Majzner R, Mackall C. Tumor antigen escape from CAR T-cell therapy. Cancer Discov 2018;8(10):1219–1226.Jacoby E, Nguyen S, Fountaine T, et al. CD19 CAR immune pressure induces B-precusor acute lymphoblastic leukaemia lineage switch exposing inherent leukaemic plasticity. Nat Commun 2016; 7:12320.