ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/005/SRR6007315/SRR6007315.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/006/SRR6007316/SRR6007316.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/003/SRR6007323/SRR6007323.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/002/SRR6007322/SRR6007322.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/004/SRR6007324/SRR6007324.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/009/SRR6007319/SRR6007319.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/001/SRR6007321/SRR6007321.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/006/SRR6007326/SRR6007326.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/007/SRR6007317/SRR6007317.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/000/SRR6007320/SRR6007320.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/008/SRR6007318/SRR6007318.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR600/005/SRR6007325/SRR6007325.fastq.gzprimaryOK200GenomicsMoriggl, Functional Cancer Genomics, University of Veterinary Medicinehttps://www.ebi.ac.uk/ena/browser/view/PRJNA401761Homo sapiensThe transcription factors STAT5A and STAT5B are essential downstream mediators of many tyrosine kinases, particularly in hematopoietic cancers. As such, STAT5 is activated by FLT3-ITD, which is a constitutively active tyrosine kinase driving the pathogenesis of acute myeloid leukemia (AML). Since STAT5 is a critical mediator of diverse malignant properties of AML cells, direct targeting of STAT5 function is of significant clinical value. Here, we describe the novel small molecular weight inhibitor AC-4-130 that directly binds to the phosphotyrosine (pY)-binding pocket of the STAT5 SH2 domain, thereby disrupting STAT5 activation, dimerization, nuclear translocation, and STAT5-dependent induction of gene transcription. AC-4-130 substantially impaired the proliferation and clonogenic growth of human AML cell lines and primary FLT3-ITD+ AML patient cells in vitro and in vivo. Importantly, AC-4-130 synergistically increased the cytotoxicity of the JAK1/2 inhibitor Ruxolitinib and the p300/pCAF inhibitor Garcinol. In summary, we report the development and preclinical evaluation of a novel, potent STAT5 SH2 domain inhibitor that can efficiently block pathological levels of STAT5 activity in AML. The synergistic effects of AC-4-130 tyrosine kinase inhibitors as well as emerging treatment strategies provide new opportunities for combinatorial treatment of leukemia and potentially other cancers. Overall design: MV4-11 and MOLM-13 cells were treated in triplicates with 5 µM AC-4-130 or DMSO (Ctrl)ENAfalseHomo sapiensPharmacologic inhibition of STAT5 in AML2025-09-242018-09-03PRJNA401761GSE1035109606