Peripheral blood marker of residual acute leukemia after hematopoietic cell transplantation using multi-plex digital droplet PCR.

MedStar author(s):
Citation: Frontiers in Immunology. 13:999298, 2022.PMID: 36248870Department: MedStar Georgetown University Hospital/MedStar Washington Hospital Center | Pediatrics ResidencyForm of publication: Journal ArticleMedline article type(s): Journal ArticleSubject headings: *Hematopoietic Stem Cell Transplantation | *Leukemia, Myeloid, Acute | Biomarkers | Disease Progression | Hematopoietic Stem Cell Transplantation/mt [Methods] | Humans | Leukemia, Myeloid, Acute/di [Diagnosis] | Leukemia, Myeloid, Acute/ge [Genetics] | Leukemia, Myeloid, Acute/th [Therapy] | Neoplasm, Residual/ge [Genetics] | Polymerase Chain Reaction/mt [Methods] | Recurrence | RNA | RNA, MessengerYear: 2022ISSN:
  • 1664-3224
Name of journal: Frontiers in immunologyAbstract: Background: Relapse remains the primary cause of death after hematopoietic cell transplantation (HCT) for acute leukemia. The ability to identify minimal/measurable residual disease (MRD) via the blood could identify patients earlier when immunologic interventions may be more successful. We evaluated a new test that could quantify blood tumor mRNA as leukemia MRD surveillance using droplet digital PCR (ddPCR).Methods: The multiplex ddPCR assay was developed using tumor cell lines positive for the tumor associated antigens (TAA: WT1, PRAME, BIRC5), with homeostatic ABL1. On IRB-approved protocols, RNA was isolated from mononuclear cells from acute leukemia patients after HCT (n = 31 subjects; n = 91 specimens) and healthy donors (n = 20). ddPCR simultaneously quantitated mRNA expression of WT1, PRAME, BIRC5, and ABL1 and the TAA/ABL1 blood ratio was measured in patients with and without active leukemia after HCT.Results: Tumor cell lines confirmed quantitation of TAAs. In patients with active acute leukemia after HCT (MRD+ or relapse; n=19), the blood levels of WT1/ABL1, PRAME/ABL1, and BIRC5/ABL1 exceeded healthy donors (p<0.0001, p=0.0286, and p=0.0064 respectively). Active disease status was associated with TAA positivity (1+ TAA vs 0 TAA) with an odds ratio=10.67, (p=0.0070, 95% confidence interval 1.91 - 59.62). The area under the curve is 0.7544. Changes in ddPCR correlated with disease response captured on standard of care tests, accurately denoting positive or negative disease burden in 15/16 (95%). Of patients with MRD+ or relapsed leukemia after HCT, 84% were positive for at least one TAA/ABL1 in the peripheral blood. In summary, we have developed a new method for blood MRD monitoring of leukemia after HCT and present preliminary data that the TAA/ABL1 ratio may may serve as a novel surrogate biomarker for relapse of acute leukemia after HCT. Copyright © 2022 Stanojevic, Grant, Vesely, Knoblach, Kanakry, Nazarian, Panditharatna, Panchapakesan, Gress, Holter-Chakrabarty and Williams.All authors: Grant M, Gress RE, Holter-Chakrabarty J, Kanakry CG, Knoblach S, Nazarian J, Panchapakesan K, Panditharatna E, Stanojevic M, Vesely SK, Williams KMFiscal year: FY2023Digital Object Identifier: Date added to catalog: 2022-10-27
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Journal Article MedStar Authors Catalog Article 36248870 Available 36248870

Background: Relapse remains the primary cause of death after hematopoietic cell transplantation (HCT) for acute leukemia. The ability to identify minimal/measurable residual disease (MRD) via the blood could identify patients earlier when immunologic interventions may be more successful. We evaluated a new test that could quantify blood tumor mRNA as leukemia MRD surveillance using droplet digital PCR (ddPCR).

Methods: The multiplex ddPCR assay was developed using tumor cell lines positive for the tumor associated antigens (TAA: WT1, PRAME, BIRC5), with homeostatic ABL1. On IRB-approved protocols, RNA was isolated from mononuclear cells from acute leukemia patients after HCT (n = 31 subjects; n = 91 specimens) and healthy donors (n = 20). ddPCR simultaneously quantitated mRNA expression of WT1, PRAME, BIRC5, and ABL1 and the TAA/ABL1 blood ratio was measured in patients with and without active leukemia after HCT.

Results: Tumor cell lines confirmed quantitation of TAAs. In patients with active acute leukemia after HCT (MRD+ or relapse; n=19), the blood levels of WT1/ABL1, PRAME/ABL1, and BIRC5/ABL1 exceeded healthy donors (p<0.0001, p=0.0286, and p=0.0064 respectively). Active disease status was associated with TAA positivity (1+ TAA vs 0 TAA) with an odds ratio=10.67, (p=0.0070, 95% confidence interval 1.91 - 59.62). The area under the curve is 0.7544. Changes in ddPCR correlated with disease response captured on standard of care tests, accurately denoting positive or negative disease burden in 15/16 (95%). Of patients with MRD+ or relapsed leukemia after HCT, 84% were positive for at least one TAA/ABL1 in the peripheral blood. In summary, we have developed a new method for blood MRD monitoring of leukemia after HCT and present preliminary data that the TAA/ABL1 ratio may may serve as a novel surrogate biomarker for relapse of acute leukemia after HCT. Copyright © 2022 Stanojevic, Grant, Vesely, Knoblach, Kanakry, Nazarian, Panditharatna, Panchapakesan, Gress, Holter-Chakrabarty and Williams.

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