15‐color highly sensitive flow cytometry assay for post <scp>anti‐CD19</scp> targeted therapy (<scp>anti‐CD19‐CAR‐T</scp> and blinatumomab) measurable residual disease assessment in B‐lymphoblastic leukemia/lymphoma: Real‐world applicability and challenges

Chatterjee, Gaurav; Dhende, Priyanka; Raj, Simpy; Shetty, Vruksha; Ghogale, Sitaram; Deshpande, Nilesh; Girase, Karishma; Patil, Jagruti; Kalra, Aastha; Narula, Gaurav; Dalvi, Kajal; Dhamne, Chetan; Moulik, Nirmalya Roy; Rajpal, Sweta; Patkar, Nikhil V.; Banavali, Shripad; Gujral, Sumeet; Subramanian, Papagudi G.; Tembhare, Prashant R.

doi:10.1111/ejh.14102

Cited by 4 publications

(1 citation statement)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Unfortunately, the cohort did not contain patients with CD19-negative MRD. Chatterjee et al [123] reported a 15-color panel using CD19, HLA-DR, CD22, and CD24 as B-cell markers, together with common maturation and aberrancy markers in combination with a gating strategy without use of CD19. This protocol allowed MRD assessment with a LOD of 0.0004% and LLOQ of 0.001% in patients with CD19-negative MRD when 5 × 10 6 cells are analyzed.…”

Section: Challenges and Novel Approaches For Flowcytometric Mrd Asses...mentioning

confidence: 99%

The Evolving Landscape of Flowcytometric Minimal Residual Disease Monitoring in B-Cell Precursor Acute Lymphoblastic Leukemia

Verbeek,

van der Velden

2024

IJMS

View full text Add to dashboard Cite

Detection of minimal residual disease (MRD) is a major independent prognostic marker in the clinical management of pediatric and adult B-cell precursor Acute Lymphoblastic Leukemia (BCP-ALL), and risk stratification nowadays heavily relies on MRD diagnostics. MRD can be detected using flow cytometry based on aberrant expression of markers (antigens) during malignant B-cell maturation. Recent advances highlight the significance of novel markers (e.g., CD58, CD81, CD304, CD73, CD66c, and CD123), improving MRD identification. Second and next-generation flow cytometry, such as the EuroFlow consortium’s eight-color protocol, can achieve sensitivities down to 10−5 (comparable with the PCR-based method) if sufficient cells are acquired. The introduction of targeted therapies (especially those targeting CD19, such as blinatumomab or CAR-T19) introduces several challenges for flow cytometric MRD analysis, such as the occurrence of CD19-negative relapses. Therefore, innovative flow cytometry panels, including alternative B-cell markers (e.g., CD22 and CD24), have been designed. (Semi-)automated MRD assessment, employing machine learning algorithms and clustering tools, shows promise but does not yet allow robust and sensitive automated analysis of MRD. Future directions involve integrating artificial intelligence, further automation, and exploring multicolor spectral flow cytometry to standardize MRD assessment and enhance diagnostic and prognostic robustness of MRD diagnostics in BCP-ALL.

show abstract

Section: Challenges and Novel Approaches For Flowcytometric Mrd Asses...mentioning

confidence: 99%

The Evolving Landscape of Flowcytometric Minimal Residual Disease Monitoring in B-Cell Precursor Acute Lymphoblastic Leukemia

Verbeek,

van der Velden

2024

IJMS

View full text Add to dashboard Cite

show abstract

Bulk lysis procedures alter target cell population counts

Rico,

Salvia,

Ward

et al. 2024

Cytometry Pt A

View full text Add to dashboard Cite

To achieve high‐sensitivity cell measurements (<1 in 105 cells) by flow cytometry (FCM), the minimum number of acquired cells must be considered and conventional immunophenotyping protocols fall short of these numbers. The bulk lysis (BL) assay is a standardized erythrocyte lysing approach that allows the analysis of the millions of cells required for high‐sensitivity measurable residual disease (MRD) detection. However, this approach has been associated with significant cell loss, along with potential over or underestimates of rare cells when using this method. The aim of this study was to evaluate bulk lysis protocols and compare them with minimal sample perturbation (MSP) protocols, which are reported to better preserve the native cellular state and avoid significant cell loss due to washing steps. To achieve this purpose, we first generated an MRD model by spiking fresh peripheral blood with K562 cells, stably expressing EGFP, at known percentages of EGFP positive cells to leukocytes. Samples were then prepared with BL and MSP protocols and analyzed using FCM. For all percentages of K562 cells established and evaluated, a significant decrease of this population was detected in BL samples compared with MSP samples, even at low K562 cell percentages. Significant decreases for non‐necrotic cells were also observed in BL samples relative to MSP samples. In conclusion, the evaluation of the potential effects of BL protocols in obtaining the final count is of great interest, especially for over‐ or under‐estimation of target cells, as in the case of measurable residual disease. Since conventional flow cytometry or minimal sample perturbation assays fall short in obtaining the minimum numbers required to reach high sensitivity measurements, significant efforts may be needed to improve bulk lysis solution reagents.

show abstract

Utility of leukocyte‐associated immunoglobulin‐like receptor‐1 (CD305) in flow cytometric detection of minimal bone marrow involvement by B‐cell non‐Hodgkin lymphoma

Singh,

Patil,

Ghogale

et al. 2024

Cytometry Part B Clinical

View full text Add to dashboard Cite

Multicolor flow cytometry (MFC) is crucial in detecting occult or minimal bone marrow (BM) involvement by non‐Hodgkin lymphomas (NHL), which may not be detected using trephine biopsy or imaging studies. Detection of low‐level BM involvement can be challenging without definite immunophenotypic aberrancies. We studied the utility of CD305 in MFC detection of minimal BM involvement by B‐NHL, especially in the absence of aberrancies by commonly used markers. The study included 1084 consecutive BM samples submitted for the staging of B‐NHLs (excluding CLL) over two years. Samples were studied for morphological, immunophenotypic, and histopathological assessment. MFC studies were performed using 10–13 color MFC, including CD305‐antibody (clone, DX26). Minimal BM involvement was defined with a cutoff of ≤10% lymphoma cells in viable cells on MFC assessment. Of 1084, 148 samples revealed overt morphological involvement by B‐NHL and were excluded from analysis. BM samples of 172/936 patients were morphologically negative but revealed involvement using MFC independently. Corresponding trephine biopsy involvement was detected in only 79/172 (45.9%) patients. On MFC, 23/172 samples showed BM involvement with >10% lymphoma cells, and 149/172 (86.6%) samples revealed minimal involvement. In 54/149 (36.24%) samples, lymphoma cells were detected only with aberrant loss of CD305 expression. In 78 of the remaining 95 samples (82.1%), it provided an immunophenotypic aberrancy addition to other markers and supported the results. CD305 is a highly useful marker in the flow cytometric assessment of minimal BM involvement by B‐NHL. MFC is a superior modality to trephine biopsy in detecting low‐level BM involvement.

show abstract

15‐color highly sensitive flow cytometry assay for post anti‐CD19 targeted therapy (anti‐CD19‐CAR‐T and blinatumomab) measurable residual disease assessment in B‐lymphoblastic leukemia/lymphoma: Real‐world applicability and challenges

Cited by 4 publications

References 52 publications

The Evolving Landscape of Flowcytometric Minimal Residual Disease Monitoring in B-Cell Precursor Acute Lymphoblastic Leukemia

The Evolving Landscape of Flowcytometric Minimal Residual Disease Monitoring in B-Cell Precursor Acute Lymphoblastic Leukemia

Bulk lysis procedures alter target cell population counts

Utility of leukocyte‐associated immunoglobulin‐like receptor‐1 (CD305) in flow cytometric detection of minimal bone marrow involvement by B‐cell non‐Hodgkin lymphoma

Contact Info

Product

Resources

About