An ShRNA Screen Identifies MEIS1 as a Driver of Malignant Peripheral Nerve Sheath Tumors

Ami V Patel; Katherine E Chaney; Kwangmin Choi; David A Largaespada; Ashish R Kumar; Nancy Ratner

doi:10.1016/j.ebiom.2016.06.007

An ShRNA Screen Identifies MEIS1 as a Driver of Malignant Peripheral Nerve Sheath Tumors

EBioMedicine. 2016 Jul:9:110-119. doi: 10.1016/j.ebiom.2016.06.007. Epub 2016 Jun 4.

Authors

Ami V Patel¹, Katherine E Chaney¹, Kwangmin Choi¹, David A Largaespada², Ashish R Kumar³, Nancy Ratner⁴

Affiliations

¹ Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital, Department of Pediatrics, University of Cincinnati, Cincinnati, OH 45229-0713, United States.
² Department of Pediatrics, Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, United States.
³ Division of Bone Marrow Transplantation & Immune Deficiency, Cincinnati Children's Hospital, Department of Pediatrics, University of Cincinnati, Cincinnati, OH 45229-0713, United States.
⁴ Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital, Department of Pediatrics, University of Cincinnati, Cincinnati, OH 45229-0713, United States. Electronic address: [email protected].

Abstract

Malignant peripheral nerve sheath tumors (MPNST) are rare soft tissue sarcomas that are a major source of mortality in neurofibromatosis type 1 (NF1) patients. To identify MPNST driver genes, we performed a lentiviral short hairpin (sh) RNA screen, targeting all 130 genes up-regulated in neurofibroma and MPNSTs versus normal human nerve Schwann cells. NF1 mutant cells show activation of RAS/MAPK signaling, so a counter-screen in RAS mutant carcinoma cells was performed to exclude common RAS-pathway driven genes. We identified 7 genes specific for survival of MPSNT cells, including MEIS1. MEIS1 was frequently amplified or hypomethylated in human MPSNTs, correlating with elevated MEIS1 gene expression. In MPNST cells and in a genetically engineered mouse model, MEIS1 expression in developing nerve glial cells was necessary for MPNST growth. Mechanistically, MEIS1 drives MPNST cell growth via the transcription factor ID1, thereby suppressing expression of the cell cycle inhibitor p27(Kip) and maintaining cell survival.

Keywords: G0/G1 arrest; ID1; MEIS1; MPNST; RNAi; Sarcoma; p27(Kip).

MeSH terms

Animals
Cell Line, Tumor
Cell Survival
Cyclin-Dependent Kinase Inhibitor p16 / deficiency
Cyclin-Dependent Kinase Inhibitor p16 / genetics
Cyclin-Dependent Kinase Inhibitor p27 / metabolism
Cyclins / genetics
Cyclins / metabolism
Disease Models, Animal
G1 Phase Cell Cycle Checkpoints
Genotype
Homeodomain Proteins / antagonists & inhibitors
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism*
Humans
Mice
Mice, Inbred C57BL
Mice, Knockout
Myeloid Ecotropic Viral Integration Site 1 Protein
Neoplasm Proteins / antagonists & inhibitors
Neoplasm Proteins / genetics
Neoplasm Proteins / metabolism*
Nerve Sheath Neoplasms / genetics
Nerve Sheath Neoplasms / metabolism
Nerve Sheath Neoplasms / mortality
Nerve Sheath Neoplasms / pathology*
Neurofibromatosis 1 / metabolism
Neurofibromatosis 1 / mortality
Neurofibromatosis 1 / pathology
Plasmids / genetics
Plasmids / metabolism
RNA Interference
RNA, Small Interfering / metabolism*
Schwann Cells / cytology
Schwann Cells / metabolism
Signal Transduction
Soft Tissue Neoplasms / genetics
Soft Tissue Neoplasms / metabolism
Soft Tissue Neoplasms / mortality
Soft Tissue Neoplasms / pathology*

Substances

CCNE2 protein, human
Cyclin-Dependent Kinase Inhibitor p16
Cyclins
Homeodomain Proteins
MEIS1 protein, human
Meis1 protein, mouse
Myeloid Ecotropic Viral Integration Site 1 Protein
Neoplasm Proteins
RNA, Small Interfering
Cyclin-Dependent Kinase Inhibitor p27

Abstract

MeSH terms

Substances

Grants and funding