Bi-allelic Loss of CDKN2A Initiates Melanoma Invasion via BRN2 Activation

Hanlin Zeng; Aparna Jorapur; A Hunter Shain; Ursula E Lang; Rodrigo Torres; Yuntian Zhang; Andrew S McNeal; Thomas Botton; Jue Lin; Matthew Donne; Ingmar N Bastian; Richard Yu; Jeffrey P North; Laura Pincus; Beth S Ruben; Nancy M Joseph; Iwei Yeh; Boris C Bastian; Robert L Judson

doi:10.1016/j.ccell.2018.05.014

Bi-allelic Loss of CDKN2A Initiates Melanoma Invasion via BRN2 Activation

Cancer Cell. 2018 Jul 9;34(1):56-68.e9. doi: 10.1016/j.ccell.2018.05.014.

Authors

Hanlin Zeng¹, Aparna Jorapur¹, A Hunter Shain², Ursula E Lang², Rodrigo Torres¹, Yuntian Zhang¹, Andrew S McNeal¹, Thomas Botton², Jue Lin³, Matthew Donne⁴, Ingmar N Bastian², Richard Yu⁵, Jeffrey P North⁶, Laura Pincus⁶, Beth S Ruben⁷, Nancy M Joseph⁸, Iwei Yeh², Boris C Bastian², Robert L Judson⁹

Affiliations

¹ Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA 94143, USA; Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA.
² Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA 94143, USA; Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA; Department of Pathology, University of California San Francisco, San Francisco, CA 94115, USA.
³ Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94143, USA.
⁴ Department of Anatomy, University of California San Francisco, San Francisco, CA 94143, USA.
⁵ Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA 94143, USA; Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA; Department of Pathology, University of California San Francisco, San Francisco, CA 94115, USA; Faculty of Medicine, University of British Columbia, Vancouver, BC V6T1Z3, Canada.
⁶ Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA; Department of Pathology, University of California San Francisco, San Francisco, CA 94115, USA.
⁷ Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA; Department of Pathology, University of California San Francisco, San Francisco, CA 94115, USA; Palo Alto Medical Foundation, Palo Alto, CA 94301, USA.
⁸ Department of Pathology, University of California San Francisco, San Francisco, CA 94115, USA.
⁹ Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA 94143, USA; Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA. Electronic address: robert.judson@ucsf.edu.

Abstract

Loss of the CDKN2A tumor suppressor is associated with melanoma metastasis, but the mechanisms connecting the phenomena are unknown. Using CRISPR-Cas9 to engineer a cellular model of melanoma initiation from primary human melanocytes, we discovered that a lineage-restricted transcription factor, BRN2, is downstream of CDKN2A and directly regulated by E2F1. In a cohort of melanocytic tumors that capture distinct progression stages, we observed that CDKN2A loss coincides with both the onset of invasive behavior and increased BRN2 expression. Loss of the CDKN2A protein product p16^INK4A permitted metastatic dissemination of human melanoma lines in mice, a phenotype rescued by inhibition of BRN2. These results demonstrate a mechanism by which CDKN2A suppresses the initiation of melanoma invasion through inhibition of BRN2.

Keywords: BRN2; CDKN2A; CRISPR engineering; E2F1; invasion; melanocytes; melanoma.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
Cell Movement*
Cyclin-Dependent Kinase Inhibitor p16 / genetics*
Cyclin-Dependent Kinase Inhibitor p16 / metabolism
E2F1 Transcription Factor / genetics
E2F1 Transcription Factor / metabolism
Female
Gene Expression Regulation, Neoplastic
Homeodomain Proteins / genetics*
Homeodomain Proteins / metabolism
Humans
Loss of Heterozygosity*
Lung Neoplasms / genetics*
Lung Neoplasms / metabolism
Lung Neoplasms / secondary
Male
Melanocytes / metabolism*
Melanocytes / pathology
Melanoma / genetics*
Melanoma / metabolism
Melanoma / secondary
Mice, Inbred NOD
Neoplasm Invasiveness
POU Domain Factors / genetics*
POU Domain Factors / metabolism
Point Mutation
Proto-Oncogene Proteins B-raf / genetics
Signal Transduction
Skin Neoplasms / genetics*
Skin Neoplasms / metabolism
Skin Neoplasms / pathology
Transcriptional Activation*

Substances

CDKN2A protein, human
Cyclin-Dependent Kinase Inhibitor p16
E2F1 Transcription Factor
E2F1 protein, human
Homeodomain Proteins
POU Domain Factors
transcription factor Brn-2
BRAF protein, human
Proto-Oncogene Proteins B-raf

Abstract

Publication types

MeSH terms

Substances

Grants and funding