bims-catcri Biomed News
on CAR-T cells, CRISPR and cancer
Issue of 2022–08–14
thirteen papers selected by
Lisa Dwane, AstraZeneca
  1. CAR T-Cell-Based gene therapy for cancers: new perspectives, challenges, and clinical developments.
  2. The landscape of chimeric antigen receptor T cell therapy in breast cancer: Perspectives and outlook.
  3. Immunotherapy and immunoengineering for breast cancer; a comprehensive insight into CAR-T cell therapy advancements, challenges and prospects.
  4. Advances and challenges of CAR T therapy and suitability of animal models (Review).
  5. Potent preclinical activity of FLT3-directed chimeric antigen receptor T cell immunotherapy against FLT3-mutant acute myeloid leukemia and KMT2A-rearranged acute lymphoblastic leukemia.
  6. The Impact of Race, Ethnicity, and Obesity on CAR T-cell Therapy Outcomes.
  7. Novel treatment approaches for HER2 positive solid tumors (excluding breast cancer).
  8. Programmable Attenuation of Antigenic Sensitivity for a Nanobody-Based EGFR Chimeric Antigen Receptor Through Hinge Domain Truncation.
  9. Personalised Therapies for Metastatic Triple-Negative Breast Cancer: When Target Is Not Everything.
  10. Therapeutic targeting of VEGF and/or TGF-β to enhance anti-PD-(L)1 therapy: The evidence from clinical trials.
  11. Generation of a CRISPR activation mouse that enables modelling of aggressive lymphoma and interrogation of venetoclax resistance.
  12. Molecular monitoring of T-cell kinetics and migration in severe neurotoxicity after real-world CD19-specific chimeric antigen receptor-T cell therapy.
  13. Single-cell ATAC-seq maps the comprehensive and dynamic chromatin accessibility landscape of CAR-T cell dysfunction.
  1. Front Immunol. 2022 ;13 925985
    CAR T-Cell-Based gene therapy for cancers: new perspectives, challenges, and clinical developments.
    Manasi P Jogalekar, Ramya Lakshmi Rajendran, Fatima Khan, Crismita Dmello, Prakash Gangadaran, Byeong-Cheol Ahn.
      Chimeric antigen receptor (CAR)-T cell therapy is a progressive new pillar in immune cell therapy for cancer. It has yielded remarkable clinical responses in patients with B-cell leukemia or lymphoma. Unfortunately, many challenges remain to be addressed to overcome its ineffectiveness in the treatment of other hematological and solidtumor malignancies. The major hurdles of CAR T-cell therapy are the associated severe life-threatening toxicities such as cytokine release syndrome and limited anti-tumor efficacy. In this review, we briefly discuss cancer immunotherapy and the genetic engineering of T cells and, In detail, the current innovations in CAR T-cell strategies to improve efficacy in treating solid tumors and hematologic malignancies. Furthermore, we also discuss the current challenges in CAR T-cell therapy and new CAR T-cell-derived nanovesicle therapy. Finally, strategies to overcome the current clinical challenges associated with CAR T-cell therapy are included as well.
    Keywords:  CAR T-cell therapy; gene therapy; hematologic malignancies; immunotherapy; solid cancers
    DOI:  https://doi.org/10.3389/fimmu.2022.925985
  2. Front Immunol. 2022 ;13 887471
    The landscape of chimeric antigen receptor T cell therapy in breast cancer: Perspectives and outlook.
    Hao Zhang, Shuangli Zhu, Wanjun Deng, Rui Li, Haiting Zhou, Huihua Xiong.
      Chimeric antigen receptor-T (CAR-T) cell therapy is a revolutionary adoptive cell therapy, which could modify and redirect T cells to specific tumor cells. Since CAR-T cell therapy was first approved for B cell-derived malignancies in 2017, it has yielded unprecedented progress in hematological tumors and has dramatically reshaped the landscape of cancer therapy in recent years. Currently, cumulative evidence has demonstrated that CAR-T cell therapy could be a viable therapeutic strategy for solid cancers. However, owing to the immunosuppressive tumor microenvironment (TME) and heterogenous tumor antigens, the application of CAR-T cell therapy against solid cancers requires circumventing more challenging obstacles. Breast cancer is characterized by a high degree of invasiveness, malignancy, and poor prognosis. The review highlights the underlying targets of CAR-T cell therapy in breast cancer, summarizes the challenges associated with CAR-T cell therapy, and proposes the strategies to overcome these challenges, which provides a novel approach to breast cancer treatment.
    Keywords:  CAR-T cell; breast cancer; chimeric antigen receptor; immunotherapy; tumor microenvironment
    DOI:  https://doi.org/10.3389/fimmu.2022.887471
  3. Cell Oncol (Dordr). 2022 Aug 09.
    Immunotherapy and immunoengineering for breast cancer; a comprehensive insight into CAR-T cell therapy advancements, challenges and prospects.
    Azam Bozorgi, Maryam Bozorgi, Mozafar Khazaei.
       BACKGROUND: Breast cancer (BC) is a highly prevalent solid cancer with a high-rise infiltration of immune cells, turning it into a significant candidate for tumor-specific immunotherapies. Chimeric antigen receptor (CAR)-T cells are emerging as immunotherapeutic tools with genetically engineered receptors to efficiently recognize and attack tumor cells that express specific target antigens. Technological advancements in CAR design have provided five generations of CAR-T cells applicable to a wide range of cancer patients while boosting CAR-T cell therapy safety. However, CAR-T cell therapy is ineffective against breast cancer because of the loss of specified antigens, the immunosuppressive nature of the tumor and CAR-T cell-induced toxicities. Next-generation CAR-T cells actively pass through the tumor vascular barriers, persist for extended periods and disrupt the tumor microenvironment (TME) to block immune escape.
    CONCLUSION: CAR-T cell therapy embodies advanced immunotherapy for BC, but further pre-clinical and clinical assessments are recommended to achieve maximized efficiency and safety.
    Keywords:  Breast cancer; CAR-T cell therapy; Immunosuppressive tumor; Target antigen; Toxicity
    DOI:  https://doi.org/10.1007/s13402-022-00700-w
  4. Mol Clin Oncol. 2022 Sep;17(3): 134
    Advances and challenges of CAR T therapy and suitability of animal models (Review).
    Xavier E Ramos-Cardona, Weichuan Luo, Sulma I Mohammed.
      Chimeric antigen receptors (CARs) recently gained momentum in cancer treatment due to their ability to promote T-cell mediated responses to a specific tumor-associated antigen. CARs are part of the adoptive cell transfer (ACT) strategies that utilize patients' T lymphocytes, genetically engineered to kill cancer cells. However, despite the therapy's success against blood-related malignancies, treating solid tumors has not reached its fullest potential yet. The reasons include the complex suppressive tumor microenvironment, mutations on cancer cells' target receptors, lethal side-effects, restricted trafficking into the tumor, suboptimal persistence in vivo and the lack of animal models that faithfully resemble human tumor's immunological responses. Currently, rodent models are used to investigate the safety and efficacy of CAR therapies. However, these models are limited in representing the human disease faithfully, fail to predict the adverse treatment events and overestimate the efficacy of the therapy. On the other hand, spontaneously developed tumors in dogs are more suited in CAR research and their efficacy has been demonstrated in a number of diseases, including lymphoma, osteosarcoma and mammary tumors. The present review discusses the design and evolution of CARs, challenges of CAR in solid tumors, human and canine clinical trials and advantages of the canine model.
    Keywords:  canine; chimeric antigen receptors therapy; dog; hematological malignancies; immunotherapy; solid tumor
    DOI:  https://doi.org/10.3892/mco.2022.2567
  5. Haematologica. 2022 Aug 11.
    Potent preclinical activity of FLT3-directed chimeric antigen receptor T cell immunotherapy against FLT3-mutant acute myeloid leukemia and KMT2A-rearranged acute lymphoblastic leukemia.
    Lisa M Niswander, Zachary T Graff, Christopher D Chien, John A Chukinas, Christina A Meadows, Lillie C Leach, Joseph P Loftus, M Eric Kohler, Sarah K Tasian, Terry J Fry.
      Chimeric antigen receptor (CAR) T cell immunotherapies targeting CD19 or CD22 induce remissions in the majority of patients with relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL), although relapse due to target antigen loss or downregulation has emerged as a major clinical dilemma. Accordingly, great interest exists in developing CAR T cells directed against alternative leukemia cell surface antigens that may help to overcome immunotherapeutic resistance. The fms-like tyrosine kinase 3 receptor (FLT3) is constitutively activated via FLT3 mutation in acute myeloid leukemia (AML) or wild-type FLT3 overexpression in KMT2A (lysine-specific methyltransferase 2A)-rearranged B-acute lymphoblastic leukemia (ALL), which are associated with poor clinical outcomes in children and adults. We developed monovalent FLT3-targeted CAR T cells (FLT3CART) and bispecific CD19xFLT3CART and assessed their anti-leukemia activity in preclinical models of FLT3-mutant AML and KMT2Arearranged infant ALL. We report robust in vitro FLT3CART-induced cytokine production and cytotoxicity against AML and ALL cell lines with minimal cross-reactivity against normal hematopoietic and non-hematopoietic tissues. We also observed potent in vivo inhibition of leukemia proliferation in xenograft models of both FLT3-mutant AML and KMT2A-rearranged ALL, including a post-tisagenlecleucel ALL-to-AML lineage switch patient-derived xenograft model pairing. We further demonstrate significant in vitro and in vivo activity of bispecific CD19xFLT3CART against KMT2A-rearranged ALL and posit that this additional approach might also diminish potential antigen escape in these high-risk leukemias. Our preclinical data credential FLT3CART as a highly effective immunotherapeutic strategy for both FLT3-mutant AML and KMT2A-R ALL that is poised for further investigation and clinical translation.
    DOI:  https://doi.org/10.3324/haematol.2022.281456
  6. Blood Adv. 2022 Aug 08. pii: bloodadvances.2022007676. [Epub ahead of print]
    The Impact of Race, Ethnicity, and Obesity on CAR T-cell Therapy Outcomes.
    Aiman J Faruqi, John A Ligon, Paul Gregory Borgman, Seth M Steinberg, Toni Foley, Lauren Little, Crystal L Mackall, Daniel W Lee, Terry J Fry, Haneen Shalabi, Jennifer N Brudno, Bonnie Yates, Lekha Mikkilineni, James N Kochenderfer, Nirali N Shah.
      Cancer outcomes with chemotherapy are inferior in patients of minority racial/ethnic groups and those with obesity. Chimeric antigen receptor (CAR) T-cell therapy has transformed outcomes for relapsed/refractory hematologic malignancies, but whether its benefits extend commensurately to racial/ethnic minorities and patients with obesity is poorly understood. With a primary focus on patients with B-cell acute lymphoblastic leukemia (B-ALL), we retrospectively evaluated the impact of demographics and obesity on CAR T-cell therapy outcomes in adult and pediatric patients with hematologic malignancies treated with CAR T-cell therapy across five phase I clinical trials at the National Cancer Institute (NCI) from 2012 to 2021. Among 139 B-ALL CAR T-cell infusions, 28.8% of patients were Hispanic, 3.6% were Black, and 29.5% were overweight/obese. No significant associations were found between race, ethnicity, or body-mass index (BMI) and complete remission rates, neurotoxicity, or overall survival. Hispanic patients were more likely to experience severe cytokine release syndrome compared to white non-Hispanic patients even after adjusting for leukemia disease burden and age (odds ratio 4.5, p = 0.001). A descriptive analysis of patients with multiple myeloma (n=24) and NHL (n=23) displayed a similar pattern to the B-ALL cohort. Our findings suggest CAR T-cell therapy may provide substantial benefit across a range of demographics characteristics, including for those populations who are at higher risk for chemotherapy resistance and relapse. However, toxicity profiles may vary. Therefore, efforts to improve access to CAR therapy for underrepresented populations and elucidate mechanisms of differential toxicity among demographic groups should be prioritized.
    DOI:  https://doi.org/10.1182/bloodadvances.2022007676
  7. Curr Opin Oncol. 2022 Aug 05.
    Novel treatment approaches for HER2 positive solid tumors (excluding breast cancer).
    Elisabeth Gaye, Nicolas Penel, Loïc Lebellec.
       PURPOSE OF THE REVIEW: Human epidermal growth factor 2 (HER2) alterations (protein overexpression, gene amplification and mutation) play a key role in oncogenesis and are more likely correlated to poorer outcome in solid tumors. We reviewed recently published studies in the last 18 months on novel treatment approaches for HER2 positive solid tumors (excluding breast cancer).
    RECENT FINDINGS: Results of clinical studies assessing anti-HER2 therapies have been recently issued.One of the most promising drugs is transtuzumab deruxtecan, an antibody-drug conjugate which demonstrated clinically meaningful activity in gastric or gastroesophageal junction cancer and colorectal cancers.Small molecules such as poziotinib, pyrotinib, neratinib, which target both epidermal growth factor receptor (EGFR) and HER2 also showed promising activity, especially in heavily pretreated ERRB2-mutated non-small cell lung cancer (NSCLC) cancer patients. Yet, these findings need to be confirmed in confirmatory randomized trials with larger cohorts.Trastuzumab-based combinations with chemotherapy or immune checkpoint inhibitors are under development with promising results, but not in all HER2 tumors. Emerging adverse events with anti-HER2 are interstitial pneumopathy and diarrhea.
    SUMMARY: Tyrosine kinase inhibitors, antibody drug conjugate in monotherapy and combinations are emerging strategies in many HER2-positive cancers; HER2 therapies are now part of standard of care of HER2-amplified gastric or gastroesophageal junction cancer. Data are pending on several unmet medical needs.
    DOI:  https://doi.org/10.1097/CCO.0000000000000873
  8. Front Immunol. 2022 ;13 864868
    Programmable Attenuation of Antigenic Sensitivity for a Nanobody-Based EGFR Chimeric Antigen Receptor Through Hinge Domain Truncation.
    Scott McComb, Tina Nguyen, Alex Shepherd, Kevin A Henry, Darin Bloemberg, Anne Marcil, Susanne Maclean, Ahmed Zafer, Rénald Gilbert, Christine Gadoury, Robert A Pon, Traian Sulea, Qin Zhu, Risini D Weeratna.
      Epidermal growth factor family receptor (EGFR) is commonly overexpressed in many solid tumors and an attractive target for chimeric antigen receptor (CAR)-T therapy, but as EGFR is also expressed at lower levels in healthy tissues a therapeutic strategy must balance antigenic responsiveness against the risk of on-target off-tumor toxicity. Herein, we identify several camelid single-domain antibodies (also known as nanobodies) that are effective EGFR targeting moieties for CARs (EGFR-sdCARs) with very strong reactivity to EGFR-high and EGFR-low target cells. As a strategy to attenuate their potent antigenic sensitivity, we performed progressive truncation of the human CD8 hinge commonly used as a spacer domain in many CAR constructs. Single amino acid hinge-domain truncation progressively decreased both EGFR-sdCAR-Jurkat cell binding to EGFR-expressing targets and expression of the CD69 activation marker. Attenuated signaling in hinge-truncated EGFR-sdCAR constructs increased selectivity for antigen-dense EGFR-overexpressing cells over an EGFR-low tumor cell line or healthy donor derived EGFR-positive fibroblasts. We also provide evidence that epitope location is critical for determining hinge-domain requirement for CARs, as hinge truncation similarly decreased antigenic sensitivity of a membrane-proximal epitope targeting HER2-CAR but not a membrane-distal EGFRvIII-specific CAR. Hinge-modified EGFR-sdCAR cells showed clear functional attenuation in Jurkat-CAR-T cells and primary human CAR-T cells from multiple donors in vitro and in vivo. Overall, these results indicate that hinge length tuning provides a programmable strategy for throttling antigenic sensitivity in CARs targeting membrane-proximal epitopes, and could be employed for CAR-optimization and improved tumor selectivity.
    Keywords:  CAR optimization; CAR-T; EGFR; cancer selectivity; cell therapy; cellular immunotherapy; hinge domain
    DOI:  https://doi.org/10.3389/fimmu.2022.864868
  9. Cancers (Basel). 2022 Jul 31. pii: 3729. [Epub ahead of print]14(15):
    Personalised Therapies for Metastatic Triple-Negative Breast Cancer: When Target Is Not Everything.
    Serena Capici, Luca Carlofrancesco Ammoni, Nicole Meli, Viola Cogliati, Francesca Fulvia Pepe, Francesca Piazza, Marina Elena Cazzaniga.
      Triple-negative breast cancer-defined by the absence of oestrogen/progesterone receptors and human epidermal growth factor receptor 2 expression-is a complex and heterogeneous type of tumour characterised by poor prognosis, aggressive behaviour and lack of effective therapeutic strategies. The identification of new biomarkers and molecular signatures is leading to development of new therapeutic strategies including immunotherapy, targeted therapy and antibody-drug conjugates (ADCs). Against a background where chemotherapy has always been considered the standard of care, evolution towards a precision medicine approach could improve TNBC clinical practice in a complex scenario, with many therapeutic options and new drugs. The aim of this review was to focus on emerging therapeutic targets and their related specific therapy, discussing available and emerging drugs, underlining differences in approval by American and European regulatory authorities and showing the future perspective in the large number of ongoing clinical trials.
    Keywords:  PI3K/AKT/mTOR; TNBC; androgen receptor; metronomic
    DOI:  https://doi.org/10.3390/cancers14153729
  10. Front Oncol. 2022 ;12 905520
    Therapeutic targeting of VEGF and/or TGF-β to enhance anti-PD-(L)1 therapy: The evidence from clinical trials.
    Linwei Li, Qinglian Wen, Ruilin Ding.
      Normalizing the tumor microenvironment (TME) is a potential strategy to improve the effectiveness of immunotherapy. Vascular endothelial growth factor (VEGF) and transforming growth factor (TGF)-β pathways play an important role in the development and function of the TME, contributing to the immunosuppressive status of TME. To inhibit VEGF and/or TGF-β pathways can restore TME from immunosuppressive to immune-supportive status and enhance sensitivity to immunotherapy such as programmed death protein-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) inhibitors. In this review, we described the existing preclinical and clinical evidence supporting the use of anti-VEGF and/or anti-TGF-β therapies to enhance cancer immunotherapy. Encouragingly, adopting anti-VEGF and/or anti-TGF-β therapies as a combination treatment with anti-PD-(L)1 therapy have been demonstrated as effective and tolerable in several solid tumors in clinical trials. Although several questions need to be solved, the clinical value of this combination strategy is worthy to be studied further.
    Keywords:  PD-1; TGF-β; VEGF; immunotherapy; review; tumor microenvironment
    DOI:  https://doi.org/10.3389/fonc.2022.905520
  11. Nat Commun. 2022 Aug 12. 13(1): 4739
    Generation of a CRISPR activation mouse that enables modelling of aggressive lymphoma and interrogation of venetoclax resistance.
    Yexuan Deng, Sarah T Diepstraten, Margaret A Potts, Göknur Giner, Stephanie Trezise, Ashley P Ng, Gerry Healey, Serena R Kane, Amali Cooray, Kira Behrens, Amy Heidersbach, Andrew J Kueh, Martin Pal, Stephen Wilcox, Lin Tai, Warren S Alexander, Jane E Visvader, Stephen L Nutt, Andreas Strasser, Benjamin Haley, Quan Zhao, Gemma L Kelly, Marco J Herold.
      CRISPR technologies have advanced cancer modelling in mice, but CRISPR activation (CRISPRa) methods have not been exploited in this context. We establish a CRISPRa mouse (dCas9a-SAMKI) for inducing gene expression in vivo and in vitro. Using dCas9a-SAMKI primary lymphocytes, we induce B cell restricted genes in T cells and vice versa, demonstrating the power of this system. There are limited models of aggressive double hit lymphoma. Therefore, we transactivate pro-survival BCL-2 in Eµ-MycT/+;dCas9a-SAMKI/+ haematopoietic stem and progenitor cells. Mice transplanted with these cells rapidly develop lymphomas expressing high BCL-2 and MYC. Unlike standard Eµ-Myc lymphomas, BCL-2 expressing lymphomas are highly sensitive to the BCL-2 inhibitor venetoclax. We perform genome-wide activation screens in these lymphoma cells and find a dominant role for the BCL-2 protein A1 in venetoclax resistance. Here we show the potential of our CRISPRa model for mimicking disease and providing insights into resistance mechanisms towards targeted therapies.
    DOI:  https://doi.org/10.1038/s41467-022-32485-9
  12. Haematologica. 2022 Aug 11.
    Molecular monitoring of T-cell kinetics and migration in severe neurotoxicity after real-world CD19-specific chimeric antigen receptor-T cell therapy.
    Susanna Carolina Berger, Boris Fehse, Nuray Akyüz, Maria Geffken, Christine Wolschke, Dietlinde Janson, Nico Gagelmann, Marlene Luther, Dominic Wichmann, Christian Frenzel, Guenther Thayssen, Anna Alegiani, Anita Badbaran, Silke Zeschke, Judith Dierlamm, Nicolaus Kröger, Francis A Ayuk.
      CD19-specific chimeric antigen receptor (CD19-CAR)-T cell therapies mediate durable responses in late-stage B-cell malignancies, but can be complicated by a potentially severe immune effector cell-associated neurotoxicity syndrome (ICANS). Despite broad efforts, the precise mechanisms of ICANS are not entirely known, and resistance to current ICANSdirected therapies (especially corticosteroids) has been observed. Recent data suggest that inflammatory cytokines and/or targeting of cerebral CD19-expressing pericytes can disrupt the blood-brain barrier and facilitate influx of immune cells, including CAR-T cells. However, specific tools for CD19-CAR-T cell analysis within often minute samples of cerebrospinal fluid (CSF) are not broadly available. Here, we applied our recently developed digital-PCR assays to monitor CD19-CAR-T cell kinetics in CSF and blood in real-world patients with neurotoxicity. Consistently, we observed a CAR-T cell enrichment within CSF in ICANS patients with further progressive accumulation despite intense corticosteroid-containing immuno-chemotherapies in a subset of patients with prolonged and therapy-resistant grade 3-4 neurotoxicity. We used next-generation T-cell receptor-β sequencing to assess the repertoire of treatment-refractory cells. Longitudinal analysis revealed a profound skewing of the T-cell receptor repertoire, which at least partly reflected selective expansion of infused Tcell clones. Interestingly, a major fraction of eventually dominating hyperexpanded T-cell clones were of non-CAR-T cell derivation. These findings hint to a role of therapy-refractory T-cell clones in severe ICANS development and prompt future systematic research to determine if CAR-T cells may serve as 'door openers' and to further characterize both CARpositive and non-CAR-T cells to interrogate the transcriptional signature of these possibly pathologic T cells.
    DOI:  https://doi.org/10.3324/haematol.2022.281110
  13. Leukemia. 2022 Aug 12.
    Single-cell ATAC-seq maps the comprehensive and dynamic chromatin accessibility landscape of CAR-T cell dysfunction.
    Penglei Jiang, Zhaoru Zhang, Yongxian Hu, Zuyu Liang, Yingli Han, Xia Li, Xin Zeng, Hao Zhang, Meng Zhu, Jian Dong, He Huang, Pengxu Qian.
      Chimeric antigen receptor T cells (CAR-T) therapy has achieved remarkable therapeutic success in treating a variety of hematopoietic malignancies. However, the high relapse rate and poor in vivo persistence, partially caused by CAR-T cell exhaustion, are still important barriers against CAR-T therapy. It remains largely elusive on the mechanisms of CAR-T exhaustion and how to attenuate exhaustion to achieve better therapeutic efficacy. In this study, we initially observed that CAR-T cells showed rapid differentiation and increased exhaustion after co-culture with tumor cells in vitro, and then performed single-cell ATAC-seq to depict the comprehensive and dynamic landscape of chromatin accessibility of CAR-T cells during tumor cell stimulation. Analyses of differential chromatin accessible regions and motif accessibility revealed that TFs were distinct in each cell type and reconstituted a coordinated regulatory network to drive CAR-T exhaustion. Furthermore, we performed scATAC-seq in patient-derived CAR-T cells and identified BATF and IRF4 as pivotal regulators in CAR-T cell exhaustion. Finally, knockdown of BATF or IRF4 enhanced the killing ability, inhibited exhaustion, and prolonged the persistence of CAR-T cells in vivo. Together, our study unraveled the epigenetic regulatory mechanisms of CAR-T exhaustion and provided new insights into CAR-T engineering to achieve better clinical treatment benefits.
    DOI:  https://doi.org/10.1038/s41375-022-01676-0
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