Calcitonin receptor-like (CALCRL) is a G-protein-coupled neuropeptide receptor involved in the regulation of blood pressure, angiogenesis, cell proliferation, and apoptosis, and is currently emerging as a novel target for the treatment of migraine. This study characterizes the role of CALCRL in acute myeloid leukemia (AML). We analyzed CALCRL expression in collectively more than 1500 well-characterized AML patients from five international cohorts (AMLCG, HOVON, TCGA, Leucegene, and UKM) and evaluated associations with survival. In the AMLCG analytic cohort, increasing transcript levels of CALCRL were associated with decreasing complete remission rates (71.5%, 53.7%, 49.6% for low, intermediate, high CALCRL expression), 5-year overall (43.1%, 26.2%, 7.1%), and event-free survival (29.9%, 15.8%, 4.7%) (all P < 0.001). CALCRL levels remained associated with all endpoints on multivariable regression analyses. The prognostic impact was confirmed in all validation sets. Genes highly expressed in CALCRLhigh AML were significantly enriched in leukemic stem cell signatures and CALCRL levels were positively linked to the engraftment capacity of primary patient samples in immunocompromised mice. CRISPR-Cas9-mediated knockout of CALCRL significantly impaired colony formation in human myeloid leukemia cell lines. Overall, our study demonstrates that CALCRL predicts outcome beyond existing risk factors and is a potential therapeutic target in AML.
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This work is dedicated to the memory of Professor Thomas Büchner. We thank all patients and clinicians participating in the trials. The help of Irina Arnhold, Mirco Witte, and Hans-Joachim Schnittler with TMA staining and analysis is gratefully acknowledged. The statistical analysis was partially funded by the José Carreras Foundation (DJCLS H 09/01f to DG). The Leucegene project is supported by the Government of Canada through Genome Canada and the Ministère de l’économie, de l’innovation et des exportations du Québec through Génome Québec, with supplementary funds from AmorChem. GS and JH are recipients of research chairs from the Canada Research Chair program and Industrielle-Alliance (Université de Montréal), respectively. The Banque de cellules leucémiques du Québec (BCLQ) is supported by grants from the Cancer Research Network of the Fonds de recherche du Québec-Santé. CP is supported by the German Cancer Aid (70111531). SKB is supported by Leukemia & Blood Cancer New Zealand and the family of Marijanna Kumerich. GL and WEB are supported by the German Research Foundation (DFG EXC 1003, Cluster of excellence “Cells in Motion”). TH and KS are supported by the Wilhelm Sander Foundation (2013.086.1 to TH and KS and 2013.086.2 to TH). TH is supported by the Physician Scientists Grant (G-509200-004) from the Helmholtz Zentrum München. C Schliemann and LA are supported by the Innovative Medical Research Fund of the University of Münster Medical School (SC211008 to CS and AN111813 to LA).