Expression of CD304/neuropilin‐1 in adult b‐cell lymphoblastic leukemia/lymphoma and its utility for the measurable residual disease assessment

Abstract

Introduction Many new markers are being evaluated to increase the sensitivity and applicability of multicolor flow cytometry (MFC)‐based measurable residual disease (MRD) monitoring. However, most of the studies are limited to childhood B‐cell lymphoblastic leukemia/lymphoma (B‐ALL), and reports in adult B‐ALL are extremely scarce and limited to small cohorts. We studied the expression of CD304/neuropilin‐1 in a large cohort of adult B‐ALL patients and evaluated its practical utility in MFC‐based MRD analysis. Methods CD304 was studied in blasts from adult B‐ALL patients and normal precursor B cells (NPBC) from non‐B‐ALL bone marrow samples using MFC. CD304 expression intensity and pattern were studied with normalized‐mean fluorescent intensity (nMFI) and coefficient of variation of immunofluorescence (CVIF), respectively. MFC‐based MRD was performed at end of induction (EOI; day‐35), end of consolidation (EOC; day 78‐80), and subsequent follow‐up (SFU) time points. Results CD304 was positive in 120/214(56.07%) and was significantly associated with BCR‐ABL1 fusion (P = .001). EOI‐MRD and EOC‐MRD were positive in 129/214(60.3%) and 50/81(61.72%), respectively. CD304 was positive in a significant percentage of EOI (48%, 62/129) and EOC (52%, 26/50) MRD‐positive B‐ALL samples. Its expression was retained, lost, and gained in 73.7%, 26.3%, and 11.3% of EOI‐MRD and 85.7%, 14.3%, and none of EOC‐MRD samples, respectively. Low‐level MRD (<0.01%) was detectable in 34 of all (EOI + EOC + SFU = 189) MRD‐positive samples, and CD304 was found useful in 50% of these samples. Conclusion CD304 is commonly expressed in adult B‐ALL and clearly distinguish B‐ALL blasts from normal precursor B cells. It is a stable MRD marker and distinctly useful in the detection of MFC‐based MRD monitoring, especially in high‐sensitivity MRD assay.

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

“…We designed a 15‐color BMRD assay incorporating markers based on our previous experiences and published literature review 2,13,28,39,40 . Details of the antibody panel are given in Table 1.…”

“…We designed a 15-color BMRD assay incorporating markers based on our previous experiences and published literature review. 2,13,28,39,40 Details of the antibody panel are given in Table 1. The purpose of antibodies included in this panel can be grouped into: (control samples) bi-weekly and also with the freshly prepared cocktail at the end of 6th week.…”

Minimal Residual Disease in the Management of B-Cell Acute Lymphoblastic Leukemia: A Systematic Review of Studies from Indian Settings

“…The panel of markers identified in these studies includes CD10, CD13, CD19, CD20, CD23, CD34, CD66c, CD123, CD200, and CD304 for the FCM analysis, and RQ-PCR was used for the BCR-ABL1 transcript characterization. A study by Chatterjee et al [32] evaluated the expression pattern of CD304 in a cohort of adult B-cell ALL patients and reported that CD304 was found to be positive in a significant percentage of EOI (62/129 [48%]) and EOC (26/50 [52%]) MRD-positive, B-cell ALL samples. CD123 has also received consideration as a marker for residual disease assessment and response evaluation in acute myeloid leukemia and B-ALL.…”

“…Regarding the timings for assessment, five studies followed EOI assessments on days 29-33 [22][23][24][25][26], except for studies by Chatterjee et al [32] and Das et al [33] that made assessments between days 35 and 40 and 30 and 35, respectively [27]. Individual studies (n = 2) that evaluated at midinduction on day 21 or after phase 1a induction were also identified [27,28].…”

Evaluation of diagnostic efficacy of NRP‐1/CD304 in hematological diseases

“…It had been verified that NRP-1/CD304 was a very useful and dependable marker for the MRD assessment of B-ALL because it was overexpressed in B-ALL cells compared with normal precursor B cells. [27][28][29][30] However, there are no comprehensive and detailed reports on the expression of NRP-1/CD304 in other common hematological diseases besides BPDCN, AML, and B-ALL, such as T-ALL, B-NHL, T/NK-cell lymphoma, and plasma cell neoplasms. Also, the application value of NRP-1/CD304 in these diseases is unknown.…”