Phenotypic Composition of Commercial Anti-CD19 CAR T Cells Affects <i>In Vivo</i> Expansion and Disease Response in Patients with Large B-cell Lymphoma

Monfrini, Chiara; Stella, Federico; Aragona, Vanessa; Magni, Martina; Ljevar, Silva; Vella, Cristina; Fardella, Eugenio; Chiappella, Annalisa; Nanetti, Francesca; Pennisi, Martina; Dodero, Anna; Guidetti, Anna; Corradini, Paolo; Carniti, Cristiana

doi:10.1158/1078-0432.ccr-22-0164

Cited by 29 publications

(25 citation statements)

References 34 publications

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“…Similar findings were reported by Monfrini et al. ( 70 ) who tested 42 patients (33 DLBCL, 8 primary mediastinal B-cell lymphomas and 1 MCL) treated with commercial anti-CD19 CAR-T cells. A unique ddPCR primer-probe assay was developed to quantify CAR vectors on genomic DNA.…”

Section: Ddpcr In Chronic Lymphoid Malignanciessupporting

confidence: 88%

“…These data were independent from the product employed. ddPCR showed a significant correlation with flow cytometry (r=0.95, p <0.0001 by Pearson correlation) with the advantage of detecting residual CAR-T cells in samples with limited cellularity and/or cryopreserved (bag-leftovers, cryopreserved BM, biopsies, cfDNA) ( 70 ). Different assays have been developed for commercial CAR-T cell monitoring.…”

Section: Ddpcr In Chronic Lymphoid Malignanciesmentioning

confidence: 99%

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Research Topic: Measurable Residual Disease in Hematologic Malignancies. Can digital droplet PCR improve measurable residual disease monitoring in chronic lymphoid malignancies?

Assanto

Giudice²,

Starza³

et al. 2023

Front. Oncol.

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Minimal/measurable residual disease (MRD) monitoring is progressively changing the management of hematologic malignancies. The possibility of detecting the persistence/reappearance of disease in patients in apparent clinical remission offers a refined risk stratification and a treatment decision making tool. Several molecular techniques are employed to monitor MRD, from conventional real-time quantitative polymerase chain reaction (RQ-PCR) to next generation sequencing and digital droplet PCR (ddPCR), in different tissues or compartments through the detection of fusion genes, immunoglobulin and T-cell receptor gene rearrangements or disease-specific mutations. RQ-PCR is still the gold standard for MRD analysis despite some limitations. ddPCR, considered the third-generation PCR, yields a direct, absolute, and accurate detection and quantification of low-abundance nucleic acids. In the setting of MRD monitoring it carries the major advantage of not requiring a reference standard curve built with the diagnostic sample dilution and of allowing to reduce the number of samples below the quantitative range. At present, the broad use of ddPCR to monitor MRD in the clinical practice is limited by the lack of international guidelines. Its application within clinical trials is nonetheless progressively growing both in acute lymphoblastic leukemia as well as in chronic lymphocytic leukemia and non-Hodgkin lymphomas. The aim of this review is to summarize the accumulating data on the use of ddPCR for MRD monitoring in chronic lymphoid malignancies and to highlight how this new technique is likely to enter into the clinical practice.

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Section: Ddpcr In Chronic Lymphoid Malignanciessupporting

confidence: 88%

Section: Ddpcr In Chronic Lymphoid Malignanciesmentioning

confidence: 99%

Research Topic: Measurable Residual Disease in Hematologic Malignancies. Can digital droplet PCR improve measurable residual disease monitoring in chronic lymphoid malignancies?

Assanto

Giudice²,

Starza³

et al. 2023

Front. Oncol.

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“…Different studies have shown clinical differences between Tisacel and Axicel; nevertheless, scarce information is available regarding the differences in the immunophenotype of CAR-T cells comparing these two products and most studies available are focused on the evaluation of the infusion products (38). With this background, in the current study immunophenotypic characterization of non-modified T cells and CAR-T cells was compared between both products at the time of peak expansion.…”

Section: Discussionmentioning

confidence: 99%

Monitoring of kinetics and exhaustion markers of circulating CAR-T cells as early predictive factors in patients with B-cell malignancies

et al. 2023

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PurposeCAR-T cell therapy has proven to be a disruptive treatment in the hematology field, however, less than 50% of patients maintain long-term response and early predictors of outcome are still inconsistently defined. Here, we aimed to optimize the detection of CD19 CAR-T cells in blood and to identify phenotypic features as early biomarkers associated with toxicity and outcomes.Experimental designIn this study, monitoring by flow cytometry and digital PCR (dPCR), and immunophenotypic characterization of circulating CAR-T cells from 48 patients treated with Tisa-cel or Axi-cel was performed.ResultsValidation of the flow cytometry reagent for the detection of CAR-T cells in blood revealed CD19 protein conjugated with streptavidin as the optimal detection method. Kinetics of CAR-T cell expansion in blood confirmed median day of peak expansion at seven days post-infusion by both flow cytometry and digital PCR. Circulating CAR-T cells showed an activated, proliferative, and exhausted phenotype at the time of peak expansion. Patients with increased expansion showed more severe CRS and ICANs. Immunophenotypic characterization of CAR-T cells at the peak expansion identified the increased expression of co-inhibitory molecules PD1 and LAG3 and reduced levels of the cytotoxicity marker CD107a as predictors of a better long-term disease control. ConclusionsThese data show the importance of CAR-T cells in vivo monitoring and identify the expression of PD1LAG3 and CD107a as early biomarkers of long-term disease control after CAR-T cell therapy.

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“…The second type is late relapse after achievement of good clinical response following CAR-T cell treatment ( 7 ). Detailed analysis of the input material and the produced CAR-T cells could help identify factors responsible for these types of treatment failures ( 8 – 10 ).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the input material quality for the production of tisagenlecleucel by multiparameter flow cytometry and its relation to the clinical outcome

Štach

Pytlík

Šmilauerová

et al. 2023

Pathol. Oncol. Res.

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Tisagenlecleucel (tisa-cel) is a CD19-specific CAR-T cell product approved for the treatment of relapsed/refractory (r/r) DLBCL or B-ALL. We have followed a group of patients diagnosed with childhood B-ALL (n = 5), adult B-ALL (n = 2), and DLBCL (n = 25) who were treated with tisa-cel under non-clinical trial conditions. The goal was to determine how the intensive pretreatment of patients affects the produced CAR-T cells, their in vivo expansion, and the outcome of the therapy. Multiparametric flow cytometry was used to analyze the material used for manufacturing CAR-T cells (apheresis), the CAR-T cell product itself, and blood samples obtained at three timepoints after administration. We present the analysis of memory phenotype of CD4/CD8 CAR-T lymphocytes (CD45RA, CD62L, CD27, CD28) and the expression of inhibitory receptors (PD-1, TIGIT). In addition, we show its relation to the patients’ clinical characteristics, such as tumor burden and sensitivity to prior therapies. Patients who responded to therapy had a higher percentage of CD8+CD45RA+CD27+ T cells in the apheresis, although not in the produced CAR-Ts. Patients with primary refractory aggressive B-cell lymphomas had the poorest outcomes which was characterized by undetectable CAR-T cell expansion in vivo. No clear correlation of the outcome with the immunophenotypes of CAR-Ts was observed. Our results suggest that an important parameter predicting therapy efficacy is CAR-Ts’ level of expansion in vivo but not the immunophenotype. After CAR-T cells’ administration, measurements at several timepoints accurately detect their proliferation intensity in vivo. The outcome of CAR-T cell therapy largely depends on biological characteristics of the tumors rather than on the immunophenotype of produced CAR-Ts.

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Phenotypic Composition of Commercial Anti-CD19 CAR T Cells Affects In Vivo Expansion and Disease Response in Patients with Large B-cell Lymphoma

Cited by 29 publications

References 34 publications

Research Topic: Measurable Residual Disease in Hematologic Malignancies. Can digital droplet PCR improve measurable residual disease monitoring in chronic lymphoid malignancies?

Research Topic: Measurable Residual Disease in Hematologic Malignancies. Can digital droplet PCR improve measurable residual disease monitoring in chronic lymphoid malignancies?

Monitoring of kinetics and exhaustion markers of circulating CAR-T cells as early predictive factors in patients with B-cell malignancies

Characterization of the input material quality for the production of tisagenlecleucel by multiparameter flow cytometry and its relation to the clinical outcome

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