Absar Muhammad, Alanazi Nawaf, Siyal Abdulaziz, Shammas Masood, Mahmood Amer, Basit Sulman, AlMukhaylid Sarah, Iqbal Zafar
Department of Pathology & Laboratory Medicine, King Abdulaziz Hospital National Guard, AlAhsa - Saudi Arabia.
Department of Pediatrics, King Abdulaziz Hospital, Al-Ahsa - Saudi Arabia.
J Popul Ther Clin Pharmacol. 2022 Jun 27;29(2):311-320. doi: 10.53555/jptcp.v29i02.4161. Epub 2022 Feb 23.
Chronic Myeloid Leukemia (CML) is initiated in the bone marrow due to the chromosomal translocation t(9;22), resulting in the fusion oncogene BCR-ABL. Tyrosine kinase inhibitors (TKIs) targeting BCR-ABL have transformed fatal CML into an almost curable disease. However, TKIs lose efficacy during disease progression, and the mechanism of CML progression remains to be fully understood. Additionally, common molecular biomarkers for CML progression are lacking. Our studies previously detected ANKRD36 (c.1183_1184 delGC and c.1187_1188 dupTT) associated exclusively with advanced phase CML. However, clinical validation of this finding was pending. Therefore, this study aimed to clinically validate mutated ANKRD36 as a novel biomarker of CML progression.
The study enrolled 124 patients in all phases of CML, recruited from Mayo Hospital and Hameed Latif Hospital in Lahore, Punjab, between January 2019 and August 2021. All response criteria were adopted from the European LeukemiaNet guideline 2020. Informed consent was obtained from all study subjects. The study was approved by scientific and ethical review committees of all participating centers.Sanger sequencing was employed to detect ANKRD36 mutations in CML patients in accelerated phase (AP) (n=11) and blast crisis (BC) (n=10), with chronic-phase CML (CP-CML) patients as controls (n=103). Samples were processed using Big Dye Terminator Cycle Sequencing Ready Reaction kits and sequenced using ABI Prism 3730 Genetic Analyzer, and sequencing using forward and reverse primers for ANKRD36.
During our study, 17% of CML patients progressed to advanced phases AP-CML n=11 (8.9%) and BC-CML n=10 (8.1%). The chronic- and advanced-phase patients showed significant difference with respect to male-to-female ratio, hemoglobin level, WBC count, and platelet count. Sanger sequencing detected ANKRD36 mutations c. 1183 1184 delGC and c. 1187 1185 dupTT exclusively in all AP- and BC-CML patients but in none of the CP-CML patients. Nevertheless, mutations status was not associated with male-to-female ratio, hemoglobin level, WBC count, and platelet count, which makes ANKRD32 as an independent predictor of early and terminal disease progression in CML.
The study confirms ANKRD36 as a novel genomic biomarker for early and late CML progression. Further prospective studies should be carried out in this regard. ANKRD36, although fully uncharacterized in humans, shows the highest expression in bone marrow, particularly myeloid cells. Functional integrated genomic studies are recommended to further explore the role of ANKRD36 in the biology and pathogenesis of CML.
慢性髓系白血病（CML）起源于骨髓中的染色体易位t(9;22)，导致融合致癌基因BCR-ABL的产生。靶向BCR-ABL的酪氨酸激酶抑制剂（TKIs）已将致命的CML转变为几乎可治愈的疾病。然而，TKIs在疾病进展过程中会失去疗效，CML进展的机制仍有待充分了解。此外，缺乏用于CML进展的常见分子生物标志物。我们之前的研究检测到ANKRD36（c.1183_1184 delGC和c.1187_1188 dupTT）仅与晚期CML相关。然而，这一发现的临床验证尚待完成。因此，本研究旨在对突变的ANKRD36作为CML进展的新型生物标志物进行临床验证。
本研究纳入了2019年1月至2021年8月期间从旁遮普省拉合尔的梅奥医院和哈米德·拉蒂夫医院招募的124例处于CML各阶段的患者。所有反应标准均采用欧洲白血病网2020年指南。所有研究对象均获得了知情同意。该研究得到了所有参与中心的科学和伦理审查委员会的批准。采用桑格测序法检测加速期（AP）（n=11）和急变期（BC）（n=10）的CML患者中的ANKRD36突变，以慢性期CML（CP-CML）患者作为对照（n=103）。使用Big Dye Terminator Cycle Sequencing Ready Reaction试剂盒处理样本，并使用ABI Prism 3730基因分析仪进行测序，使用ANKRD36的正向和反向引物进行测序。
在我们的研究期间，17%的CML患者进展至晚期，AP-CML患者11例（8.9%），BC-CML患者10例（8.1%）。慢性期和晚期患者在男女比例、血红蛋白水平、白细胞计数和血小板计数方面存在显著差异。桑格测序仅在所有AP-和BC-CML患者中检测到ANKRD36突变c.1183 1184 delGC和c.1187 1185 dupTT，而在CP-CML患者中均未检测到。然而，突变状态与男女比例、血红蛋白水平、白细胞计数和血小板计数无关，这使得ANKRD32成为CML早期和终末期疾病进展的独立预测指标。
该研究证实ANKRD36是CML早期和晚期进展的新型基因组生物标志物。对此应开展进一步的前瞻性研究。ANKRD36虽然在人类中完全未被表征，但在骨髓中，尤其是髓系细胞中表达最高。建议进行功能整合基因组研究，以进一步探索ANKRD36在CML生物学和发病机制中的作用。
