{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T15:04:35Z","timestamp":1784905475670,"version":"3.55.0"},"reference-count":31,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2023,12,22]],"date-time":"2023-12-22T00:00:00Z","timestamp":1703203200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100005315","name":"Natural Science Foundation of Ningbo Municipality","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005315","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Drug Development Research"],"published-print":{"date-parts":[[2024,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Acute myeloid leukemia (AML)\u00a0patients with FLT3 internal tandem duplication (FLT3\u2010ITD)\u00a0and DNA methyltransferase 3A (DNMT3A)\u00a0R882 double mutations had a worse prognosis compared with\u00a0AML with FLT3\u2010ITD\u00a0or DNMT3A R882 single mutation. This study was designed to explore the specific role of Calcitonin Receptor Like (CALCRL)\u00a0in AML with FLT3\u2010ITD\u00a0and DNMT3A R882 double mutations. MOLM13 cells were transduced with CRISPR knockout sgRNA constructs to establish the FTL3\u2010ITD\u00a0and DNMT3A\u2010R882\u00a0double\u2010mutated AML cell model. Quantitative real\u2010time PCR and Western blot assay were carried out to examine corresponding gene and protein expression. Methylation of CALCRL promoter was measured by methylation\u2010specific PCR (MSP).\u00a0Cell viability, colony formation, flow cytometry, and sphere formation assays were conducted to determine cell proliferation, apoptosis, and stemness. MOLM13 cells were exposed to stepwise increasing concentrations of cytarabine (Ara\u2010C) to generate MOLM13\/Ara\u2010C\u00a0cells. An in vivo AML \u00a0animal model was established, and the tumor volume and weight were recorded. TUNEL assay was adopted to examine cell apoptosis in tumor tissues. DNMT3A\u2010R882 mutation upregulated the expression of CALCRL while downregulated the DNA methylation level of CALCRL in MOLM13 cells. CALCRL knockdown greatly inhibited cell proliferation, promoted apoptosis\u00a0and repressed cell stemness, accompanied with the downregulated Oct4, SOX2, and Nanog in DNMT3A\u2010R882\u2010mutated MOLM13 cells and MOLM13\/Ara\u2010C cells. Furthermore, CALCRL knockdown restricted tumor growth and the chemoresistance of AML in vivo, as well as inducing cell apoptosis in tumor tissues. Together, these data reveal that CALCRL is a vital regulator of leukemia cell survival and resistance to chemotherapy, suggesting CALCRL as a promising therapeutic target for the treatment of FTL3\u2010ITD\u00a0and DNMT3A\u2010R882 double\u2010mutated AML.<\/jats:p>","DOI":"10.1002\/ddr.22137","type":"journal-article","created":{"date-parts":[[2023,12,22]],"date-time":"2023-12-22T08:28:49Z","timestamp":1703233729000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["CALCRL knockdown suppresses cancer stemness and chemoresistance in acute myeloid leukemia with FLT3\u2010ITD and DNM3TA\u2010R882 double mutations"],"prefix":"10.1002","volume":"85","author":[{"given":"Shanhao","family":"Tang","sequence":"first","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huiling","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lixia","family":"Sheng","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qitian","family":"Mu","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaihong","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miao","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhijuan","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"An","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-0723-7944","authenticated-orcid":false,"given":"Guifang","family":"Ouyang","sequence":"additional","affiliation":[{"name":"Department of Hematology The First Affiliated Hospital of Ningbo University Ningbo China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,12,22]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbmt.2015.07.030"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41375-019-0505-x"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2021005236"},{"issue":"3","key":"e_1_2_10_5_1","first-page":"158","article-title":"FLT3\u2010ITD compared with DNMT3A R882 mutation is a more powerful independent inferior prognostic factor in adult acute myeloid leukemia patients after allogeneic hematopoietic stem cell transplantation: A retrospective cohort study","volume":"35","author":"Ardestani M. 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