Digital PCR Improves Sensitivity and Quantification in Monitoring CAR-T Cells in B Cell Lymphoma Patients

de la Iglesia-San Sebastián, Ismael; Carbonell, Diego; Bastos-Oreiro, Mariana; Pérez-Corral, Ana; Bailén, Rebeca; Chicano, Maria; Muñiz, Paula; Monsalvo, Silvia; Escudero-Fernández, Asunción; Oarbeascoa, Gillen; Fernández-Caldas, Paula; Gómez-Centurión, Ignacio; Pion, Marjorie; Gayoso, Jorge; Anguita, Javier; Kwon, Mi; Díez-Martín, José Luis; Buño, Ismael; Martínez-Laperche, Carolina

doi:10.1016/j.jtct.2023.12.672

Cited by 1 publication

(1 citation statement)

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“…However, it is important to note that we cannot entirely rule out the possibility that in samples with positive ddPCR results, the negative FCM results may be attributed to the lack of transcription and surface expression in a significant proportion of cells carrying the transcript. A recent publication reported that when calibrated with house-keeping genes, the estimated proportion between CAR copies and CAR expressing cells may be set around 2–2.5:1 [ 14 ].…”

Section: Discussionmentioning

confidence: 99%

Unlocking Predictive Power: Quantitative Assessment of CAR-T Expansion with Digital Droplet Polymerase Chain Reaction (ddPCR)

Galli,

Viscovo,

Fosso

et al. 2024

IJMS

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Flow cytometry (FCM) and quantitative PCR (qPCR) are conventional methods for assessing CAR-T expansion, while digital droplet PCR (ddPCR) is emerging as a promising alternative. We monitored CAR-T transcript expansion in 40 B-NHL patients post-infusion of CAR-T products (axi-cel; tisa-cel; and brexu-cel) with both His-Tag FCM and ddPCR techniques. Sensitivity and predictive capacity for efficacy and safety outcomes of ddPCR were analyzed and compared with FCM. A significant correlation between CAR-T counts determined by FCM and CAR transcripts assessed by ddPCR (p < 0.001) was observed. FCM revealed median CD3+CAR+ cell counts at 7, 14, and 30 days post-infusion with no significant differences. In contrast, ddPCR-measured median copies of CAR-T transcripts demonstrated significant lower copy numbers in tisa-cel recipients compared to the other products at day 7 and day 14. Patients with a peak of CAR transcripts at day 7 exceeding 5000 copies/microg gDNA, termed “good CAR-T expanders”, were more likely to achieve a favorable response at 3 months (HR 10.79, 95% CI 1.16–100.42, p = 0.036). Good CAR-T expanders showed superior progression-free survival at 3, 6, and 12 months compared to poor CAR-T expanders (p = 0.088). Those reaching a peak higher than 5000 copies/microg gDNA were more likely to experience severe CRS and ICANS. DdPCR proves to be a practical method for monitoring CAR-T expansion, providing quantitative information that better predicts both treatment outcomes and toxicity.

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Section: Discussionmentioning

confidence: 99%