CAR T-Cell Therapy Predictive Response Markers in Diffuse Large B-Cell Lymphoma and Therapeutic Options After CART19 Failure

Caballero, A; Escribà-García, Laura; Álvarez-Fernández, Carmen; Briones, Javier

doi:10.3389/fimmu.2022.904497

Cited by 11 publications

(8 citation statements)

References 162 publications

(227 reference statements)

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“…CAR-T cell therapy has acquired an increasing role for the treatment of hematological and, more recently, autoimmune diseases. However, this therapy failed in around 50% of patients with hematological cancers [10]. Factors such as proliferation and in vivo persistence of this cell subset have been recognized as determinant elements influencing toxicity, outcome and durability of the response.…”

Section: Discussionmentioning

confidence: 99%

The challenge of standardizing CAR‐T cell monitoring: A comparison of two flow‐cytometry methods and correlation with qPCR technique

Valdivieso‐Shephard,

Matas‐Pérez,

García‐Bujalance

et al. 2024

Cytometry Pt A

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Chimeric antigen receptor (CAR) T‐cell therapy is a breakthrough in hematologic malignancies, such as acute B lymphoblastic leukemia (B‐ALL). Monitoring this treatment is recommended, although standardized protocols have not been developed yet. This work compares two flow cytometry monitoring strategies and correlates this technique with qPCR method. CAR‐T cells were detected by two different flow‐cytometry protocols (A and B) in nine blood samples from one healthy donor and five B‐ALL patients treated with Tisagenlecleucel (Kymriah®, USA). HIV‐1 viral load allowed CAR detection by qPCR, using samples from seven healthy donors and nine B‐ALL patients. CAR detection by protocol A and B did not yield statistically significant differences (1.9% vs. 11.8% CD3 + CAR+, p = 0.07). However, protocol B showed a better discrimination of the CD3 + CAR+ population. A strong correlation was observed between protocol B and qPCR (r = 0.7, p < 0.0001). CD3 + CAR+ cells were detected by flow cytometry only when HIV‐1 viral load was above 104 copies/mL. In conclusion, protocol B was the most specific flow‐cytometry procedure for the identification of CAR‐T cells and showed a high correlation with qPCR. Further efforts are needed to achieve a standardized monitoring approach.

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Section: Discussionmentioning

confidence: 99%

The challenge of standardizing CAR‐T cell monitoring: A comparison of two flow‐cytometry methods and correlation with qPCR technique

Valdivieso‐Shephard,

Matas‐Pérez,

García‐Bujalance

et al. 2024

Cytometry Pt A

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“…CAR T-cell therapy studies have shown similar relapse and resistance data to bispecific antibodies, with a loss of CD19 expression. 23 A single-center study of patients with large B-cell lymphoma (LBCL) treated with the anti-CD19 CAR T-cell therapy axicabtagene ciloleucel reported that 10 of 16 patient biopsies had either lost or had diminished CD19 expression at progression. 24 In accordance with these findings, CD19 expression was barely detectable in our single-cell biopsy analysis of a patient with DLBCL who had progressed after anti-CD19 CAR T-cell therapy.…”

Section: Discussionmentioning

confidence: 99%

Molecular assessment of intratumoral immune cell subsets and potential mechanisms of resistance to odronextamab, a CD20×CD3 bispecific antibody, in patients with relapsed/refractory B-cell non-Hodgkin lymphoma

Brouwer-Visser,

Fiaschi,

Deering

et al. 2024

J Immunother Cancer

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BackgroundPatients with relapsed/refractory B-cell non-Hodgkin lymphoma (R/R B-NHL) have a significant need for effective treatment options. Odronextamab is an Fc-silenced, human, CD20×CD3 bispecific antibody that targets CD20-expressing cells via T-cell-mediated cytotoxicity independent of T-cell/major histocompatibility complex interaction. Phase I results in patients with R/R B-NHL demonstrated that odronextamab monotherapy could achieve deep and durable responses with a generally manageable safety profile (ELM-1;NCT02290951). As part of a biomarker analysis of the same study, we investigated potential biomarkers and mechanisms of resistance to odronextamab.MethodsPatients with R/R B-NHL enrolled in ELM-1 received one time per week doses of intravenous odronextamab for 4×21 day cycles, then doses every 2 weeks thereafter. Patient tumor biopsies were obtained at baseline, on-treatment, and at progression. Immune cell markers were analyzed by immunohistochemistry, flow cytometry, single-cell RNA sequencing, and whole genome sequencing.ResultsBaseline tumor biopsies showed that almost all patients had high proportions of B cells that expressed the CD20 target antigen, whereas expression of other B-cell surface antigens (CD19, CD22, CD79b) was more variable. Responses to odronextamab in patients with diffuse large B-cell lymphoma were not related to the relative level of baseline CD20 expression, cell of origin, or high-risk molecular subtype. A potential link was observed between greater tumor programmed cell death-ligand 1 expression and increased likelihood of response to odronextamab. Similarly, a trend was observed between clinical response and increased levels of CD8 T cells and regulatory T cells at baseline. We also identified an on-treatment pharmacodynamic shift in intratumoral immune cell subsets. Finally, loss of CD20 expression through inactivating gene mutations was identified as a potential mechanism of resistance in patients who were treated with odronextamab until progression, as highlighted in two detailed patient cases reported here.ConclusionsThis biomarker analysis expands on clinical findings of odronextamab in patients with R/R B-NHL, providing verification of the suitability of CD20 as a therapeutic target, as well as evidence for potential mechanisms of action and resistance.

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“…CAR‐T cell expansion, either measured by flow cytometry or qPCR, commensurate with pretreatment tumor burden was the most significant determining factor for durable response [11] and CAR‐T memory phenotype seems to be directly correlated to activity and persistence in patients [39–44]. In FL and diffuse large B‐cell lymphoma, higher remission rates were observed in patients with a higher in vivo CAR‐T cell expansion peak and lower expression of exhaustion markers [45, 46].…”

Section: Discussionmentioning

confidence: 99%

Optimization and validation of in vivo flow cytometry chimeric antigen receptor T cell detection method using CD19his indirect staining

Zaninelli,

Meli,

Borleri

et al. 2023

Cytometry Pt A

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CD19‐targeted Chimeric Antigen Receptor T (CAR‐T) cell therapy has shown unprecedented results in patients with B cell relapsed/refractory acute lymphoblastic leukemia (R/R‐ALL) and B cell non‐Hodgkin lymphomas where no other curative options are available. In vivo monitoring of CAR‐T cell kinetics is fundamental to understand the correlation between CAR‐T cells expansion and persistence with treatment response and toxicity development. The aim of this study was to define a robust, sensitive, and universal method for CAR‐T cell detection using flow cytometry. We set‐up and compared with each other three assays for CD19 CAR‐T cell detection, all based on commercially available reagents. All methods used a recombinant human CD19 protein fragment recognized by the single chain variable fragment of the CAR construct. The two indirect staining assays (CD19his + APC‐conjugated anti‐histidine antibody and CD19bio + APC‐conjugated anti‐biotin antibody) showed better sensitivity and specificity compared to the direct staining with CD19‐FITC, and CD19his had a better cost‐effective profile. We validated CAR detection with CD19his with parallel quantitative real time PCR data and we could demonstrate a strong positive correlation. We also showed that CD19his staining can be easily included in a multicolor flow cytometry panel to achieve additional information about the cell phenotype of CAR‐T cell positive subpopulations. Finally, this method can be used for different anti‐CD19 CAR‐T cell products and for different sample sources. These data demonstrate that detection of CAR‐T cells by CD19his flow cytometry staining is a reliable, robust, and broadly applicable tool for in vivo monitoring of CAR‐T cells.This article is protected by copyright. All rights reserved.

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CAR T-Cell Therapy Predictive Response Markers in Diffuse Large B-Cell Lymphoma and Therapeutic Options After CART19 Failure

Cited by 11 publications

References 162 publications

The challenge of standardizing CAR‐T cell monitoring: A comparison of two flow‐cytometry methods and correlation with qPCR technique

The challenge of standardizing CAR‐T cell monitoring: A comparison of two flow‐cytometry methods and correlation with qPCR technique

Molecular assessment of intratumoral immune cell subsets and potential mechanisms of resistance to odronextamab, a CD20×CD3 bispecific antibody, in patients with relapsed/refractory B-cell non-Hodgkin lymphoma

Optimization and validation of in vivo flow cytometry chimeric antigen receptor T cell detection method using CD19his indirect staining

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