Validation of Flow Cytometric Phospho-STAT5 As a Diagnostic Tool for Juvenile Myelomonocytic Leukemia

Overview

Journal Blood Cancer J

Date 2013 Nov 19

PMID 24241400

Citations 14

Authors

D Hasegawa

C Bugarin

M Giordan

S Bresolin

D Longoni

C Micalizzi

U Ramenghi

A Bertaina

G Basso

F Locatelli

A Biondi

G Te Kronnie

G Gaipa

Affiliations

Soon will be listed here.

Abstract

To diagnose juvenile myelomonocytic leukemia (JMML) is sometimes challenging, because around 10% of patients lack molecular abnormalities affecting Ras-MAPK (mitogen-activated protein kinase) pathway and other diseases such as cytomegalovirus infection can mimic clinical signs of JMML. In order to validate a phospho-specific flow cytometry assay assessing phospho-signal transducer and activator of transcription factor 5 (p-STAT5) as a new diagnostic tool for JMML, we examined 22 samples from children with JMML and 47 controls. CD33+/CD34+ cells from 22 patients with JMML showed hyperphosphorylation of STAT5 induced by sub-saturating doses of granulocyte-macrophage colony-stimulating factor (GM-CSF). Using a training set of samples (11 JMML and 23 controls), we identified a threshold for p-STAT5-positive after stimulation with 0.1 ng/ml GM-CSF (17.17%) that discriminates JMML from controls. This threshold was validated in an independent series (11 JMML, 24 controls and 7 cases with diseases other than JMML) where we demonstrated that patients with JMML could be distinguished from other subjects with a sensitivity of 91% (confidence interval (CI) 59-100%) and a specificity of 87% (CI 70-96%). Positive and negative predictive values were 71% (CI 42-92%) and 96% (CI 82-100%), respectively. In conclusion, flow cytometric p-STAT5 profiling is a reliable diagnostic tool for identifying patients with JMML and can contribute to consistency of current diagnostic criteria.

Citing Articles

Efficacy of the Allosteric MEK Inhibitor Trametinib in Relapsed and Refractory Juvenile Myelomonocytic Leukemia: a Report from the Children's Oncology Group.

Stieglitz E, Lee A, Angus S, Davis C, Barkauskas D, Hall D Cancer Discov. 2024; 14(9):1590-1598.

PMID: 38867349 PMC: 11374478. DOI: 10.1158/2159-8290.CD-23-1376.

Optofluidic Flow Cytometer with In-Plane Spherical Mirror for Signal Enhancement.

Zorzi F, Bonfadini S, Aloisio L, Moschetta M, Storti F, Simoni F Sensors (Basel). 2023; 23(22).

PMID: 38005576 PMC: 10675696. DOI: 10.3390/s23229191.

Phenotypic profiling of CD34 cells by advanced flow cytometry improves diagnosis of juvenile myelomonocytic leukemia.

Bugarin C, Antolini L, Buracchi C, Matarraz S, Coliva T, Van der Velden V Haematologica. 2023; 109(2):521-532.

PMID: 37534527 PMC: 10828789. DOI: 10.3324/haematol.2023.282805.

Juvenile myelomonocytic leukemia in the molecular era: a clinician's guide to diagnosis, risk stratification, and treatment.

Wintering A, Dvorak C, Stieglitz E, Loh M Blood Adv. 2021; 5(22):4783-4793.

PMID: 34525182 PMC: 8759142. DOI: 10.1182/bloodadvances.2021005117.

Enhanced NF-κB signaling in type-2 dendritic cells at baseline predicts non-response to adalimumab in psoriasis.

Andres-Ejarque R, Ale H, Grys K, Tosi I, Solanky S, Ainali C Nat Commun. 2021; 12(1):4741.

PMID: 34362923 PMC: 8346545. DOI: 10.1038/s41467-021-25066-9.

References

Yoshimi A, Bader P, Matthes-Martin S, Stary J, Sedlacek P, Duffner U . Donor leukocyte infusion after hematopoietic stem cell transplantation in patients with juvenile myelomonocytic leukemia. Leukemia. 2005; 19(6):971-7. DOI: 10.1038/sj.leu.2403721. View

Irish J, Hovland R, Krutzik P, Perez O, Bruserud O, Gjertsen B . Single cell profiling of potentiated phospho-protein networks in cancer cells. Cell. 2004; 118(2):217-28. DOI: 10.1016/j.cell.2004.06.028. View

Platanias L . Map kinase signaling pathways and hematologic malignancies. Blood. 2003; 101(12):4667-79. DOI: 10.1182/blood-2002-12-3647. View

Puri K, Singh P, Das R, Seth R, Gupta R . Diagnostic dilemma of JMML coexisting with CMV infection. Indian J Pediatr. 2011; 78(4):485-7. DOI: 10.1007/s12098-010-0355-z. View

Gaipa G, Bugarin C, Longoni D, Cesana S, Molteni C, Faini A . Aberrant GM-CSF signal transduction pathway in juvenile myelomonocytic leukemia assayed by flow cytometric intracellular STAT5 phosphorylation measurement. Leukemia. 2008; 23(4):791-3. DOI: 10.1038/leu.2008.265. View

Nishio N, Takahashi Y, Tanaka M, Xu Y, Yoshida N, Sakaguchi H . Aberrant phosphorylation of STAT5 by granulocyte-macrophage colony-stimulating factor in infant cytomegalovirus infection mimicking juvenile myelomonocytic leukemia. Leuk Res. 2011; 35(9):1261-4. DOI: 10.1016/j.leukres.2011.04.014. View

Shannon K, OConnell P, Martin G, Paderanga D, Olson K, Dinndorf P . Loss of the normal NF1 allele from the bone marrow of children with type 1 neurofibromatosis and malignant myeloid disorders. N Engl J Med. 1994; 330(9):597-601. DOI: 10.1056/NEJM199403033300903. View

Suda T, Miura Y, Mizoguchi H, Ijima H, Eguchi M, Kaku H . Characterization of hemopoietic precursor cells in juvenile-type chronic myelocytic leukemia. Leuk Res. 1982; 6(1):43-53. DOI: 10.1016/0145-2126(82)90042-x. View

Loh M, Sakai D, Flotho C, Kang M, Fliegauf M, Archambeault S . Mutations in CBL occur frequently in juvenile myelomonocytic leukemia. Blood. 2009; 114(9):1859-63. PMC: 2738571. DOI: 10.1182/blood-2009-01-198416. View

10.

Pinkel D . Differentiating juvenile myelomonocytic leukemia from infectious disease. Blood. 1998; 91(1):365-7. View

11.

Lapidot T, Grunberger T, Vormoor J, Estrov Z, Kollet O, Bunin N . Identification of human juvenile chronic myelogenous leukemia stem cells capable of initiating the disease in primary and secondary SCID mice. Blood. 1996; 88(7):2655-64. View

12.

Toyoda H, Ido M, Hori H, Hiraiwa H, Hirayama M, Kobayashi M . A case of juvenile myelomonocytic leukemia with concomitant cytomegalovirus infection. J Pediatr Hematol Oncol. 2004; 26(9):606-8. DOI: 10.1097/01.mph.0000133599.34635.90. View

13.

Emanuel P, Bates L, Castleberry R, Gualtieri R, Zuckerman K . Selective hypersensitivity to granulocyte-macrophage colony-stimulating factor by juvenile chronic myeloid leukemia hematopoietic progenitors. Blood. 1991; 77(5):925-9. View

14.

Freedman M, Hitzler J, Bunin N, Grunberger T, Squire J . Juvenile chronic myelogenous leukemia multilineage CD34+ cells: aberrant growth and differentiation properties. Stem Cells. 1996; 14(6):690-701. DOI: 10.1002/stem.140690. View

15.

Arico M, Biondi A, Pui C . Juvenile myelomonocytic leukemia. Blood. 1997; 90(2):479-88. View

16.

Chan R, Cooper T, Kratz C, Weiss B, Loh M . Juvenile myelomonocytic leukemia: a report from the 2nd International JMML Symposium. Leuk Res. 2008; 33(3):355-62. PMC: 2692866. DOI: 10.1016/j.leukres.2008.08.022. View

17.

Kirby M, Weitzman S, Freedman M . Juvenile chronic myelogenous leukemia: differentiation from infantile cytomegalovirus infection. Am J Pediatr Hematol Oncol. 1990; 12(3):292-6. DOI: 10.1097/00043426-199023000-00007. View

18.

Sawai N, Koike K, Higuchi T, Ogami K, Oda M . Thrombopoietin enhances the production of myeloid cells, but not megakaryocytes, in juvenile chronic myelogenous leukemia. Blood. 1998; 91(11):4065-73. View

19.

Tartaglia M, Niemeyer C, Fragale A, Song X, Buechner J, Jung A . Somatic mutations in PTPN11 in juvenile myelomonocytic leukemia, myelodysplastic syndromes and acute myeloid leukemia. Nat Genet. 2003; 34(2):148-50. DOI: 10.1038/ng1156. View

20.

Simel D, Samsa G, Matchar D . Likelihood ratios with confidence: sample size estimation for diagnostic test studies. J Clin Epidemiol. 1991; 44(8):763-70. DOI: 10.1016/0895-4356(91)90128-v. View