Ability of Platelet CREB3L1 mRNA Expression Analysis to Detect Ph-Negative MPNs: Independent Confirmation in Archival Patient Samples

Background. Previously [1] mRNA of CREB3L1 gene expression level could reliably distinguish Ph-MPN from reactive hypercytosis it was found using RNA-seq analysis and subsequent quantitative RT-PCR in platelet-rich plasma. This reasonably suggests the prospect of creating a single simple test that would avoid the costly diver mutation analysis and bone marrow biopsy in the first step of Ph-MPN diagnosis.

Aims. Evaluate the diagnostic potential of the CREB3L1 mRNA expression in archival cDNA transcriptome samples of platelets from patients examined in our laboratory for suspected MPNs.

Methods. We used frozen at -20°C cDNA samples obtained from platelet RNA of patients examined between March 11 and June 30, 2015. A total of 49 samples from patients with confirmed Ph-MPNs (PV-26, ET-18 and MF-5) were included. Of these, 43 patients had the V617F JAK2 mutation (Me VAF = 33.5%), one patient had a mutation in exon 12 of JAK2 and one patient had a double mutation V617F JAK2 + CALR del. In three patients, driver mutations could not be detected (tri-negative MPN). The comparison group included samples from 47 patients also referred to the laboratory with suspected MPN, but who did not receive confirmation of this diagnosis. Measurement of the CREB3L1 mRNA expression level was performed according to the PCR-RT method described in the article [1] with minimal modifications. The calculation was performed using the ∆Сt method. Statistical analysis was performed using the R program.

Results. In all patients with MPNs, CREB3L1 mRNA expression in platelets was 62.5; 41.4-88.1 (Me;Q25-Q75). No dependence on routine hematological parameters, VAF V617F JAK2 values or clinical variant of MPNs was found in this sample. CREB3L1 mRNA platelets expression in patients with unconfirmed MPN was determined at a minimal level only in 21 patients and was significantly lower - 10.0; 5.8-12.1 (P<0.0001). The remaining patients in this group did not have CREB3L1 mRNA expression.

Summary and Conclusions. The ability of CREB3L1 mRNA expression analysis to identify different variants of Ph-MPNs in a retrospective study was independently confirmed using archival platelet cDNA samples from patients in the Siberian region.

Literature:

1. Morishita S, Yasuda H, Yamawaki S, Kawaji H, Itoh M, Edahiro Y, Imai M, Kogo Y, Tsuneda S, Ohsaka A, Hayashizaki Y, Ito M, Araki M, Komatsu N. CREB3L1 overexpression as a potential diagnostic marker of Philadelphia chromosome-negative myeloproliferative neoplasms. Cancer Sci. 2021;112(2):884-892. doi: 10.1111/cas.14763.

No relevant conflicts of interest to declare.