PMID- 32260433 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20200928 IS - 2072-6694 (Print) IS - 2072-6694 (Electronic) IS - 2072-6694 (Linking) VI - 12 IP - 4 DP - 2020 Apr 4 TI - Extracellular Vesicles Enriched with Moonlighting Metalloproteinase Are Highly Transmissive, Pro-Tumorigenic, and Trans-Activates Cellular Communication Network Factor (CCN2/CTGF): CRISPR against Cancer. LID - 10.3390/cancers12040881 [doi] LID - 881 AB - Matrix metalloproteinase 3 (MMP3) plays multiple roles in extracellular proteolysis as well as intracellular transcription, prompting a new definition of moonlighting metalloproteinase (MMP), according to a definition of protein moonlighting (or gene sharing), a phenomenon by which a protein can perform more than one function. Indeed, connective tissue growth factor (CTGF, aka cellular communication network factor 2 (CCN2)) is transcriptionally induced as well as cleaved by MMP3. Moreover, several members of the MMP family have been found within tumor-derived extracellular vesicles (EVs). We here investigated the roles of MMP3-rich EVs in tumor progression, molecular transmission, and gene regulation. EVs derived from a rapidly metastatic cancer cell line (LuM1) were enriched in MMP3 and a C-terminal half fragment of CCN2/CTGF. MMP3-rich, LuM1-derived EVs were disseminated to multiple organs through body fluid and were pro-tumorigenic in an allograft mouse model, which prompted us to define LuM1-EVs as oncosomes in the present study. Oncosome-derived MMP3 was transferred into recipient cell nuclei and thereby trans-activated the CCN2/CTGF promoter, and induced CCN2/CTGF production in vitro. TRENDIC and other cis-elements in the CCN2/CTGF promoter were essential for the oncosomal responsivity. The CRISPR/Cas9-mediated knockout of MMP3 showed significant anti-tumor effects such as the inhibition of migration and invasion of tumor cells, and a reduction in CCN2/CTGF promoter activity and fragmentations in vitro. A high expression level of MMP3 or CCN2/CTGF mRNA was prognostic and unfavorable in particular types of cancers including head and neck, lung, pancreatic, cervical, stomach, and urothelial cancers. These data newly demonstrate that oncogenic EVs-derived MMP is a transmissive trans-activator for the cellular communication network gene and promotes tumorigenesis at distant sites. FAU - Okusha, Yuka AU - Okusha Y AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. AD - Division of Molecular and Cellular Biology, Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA. FAU - Eguchi, Takanori AU - Eguchi T AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. AD - Advanced Research Center for Oral and Craniofacial Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. FAU - Tran, Manh T AU - Tran MT AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. FAU - Sogawa, Chiharu AU - Sogawa C AUID- ORCID: 0000-0002-5606-0710 AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. FAU - Yoshida, Kaya AU - Yoshida K AD - Department of Oral Healthcare Education, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8504, Japan. FAU - Itagaki, Mami AU - Itagaki M AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. AD - Research program for undergraduate students, Okayama University Dental School, Okayama 700-8525, Japan. FAU - Taha, Eman A AU - Taha EA AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. AD - Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan. AD - Department of Biochemistry, Ain Shams University Faculty of Science, Cairo 11566, Egypt. FAU - Ono, Kisho AU - Ono K AD - Department of Oral and Maxillofacial Surgery, Okayama University Hospital, Okayama 700-0914, Japan. FAU - Aoyama, Eriko AU - Aoyama E AD - Advanced Research Center for Oral and Craniofacial Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. FAU - Okamura, Hirohiko AU - Okamura H AUID- ORCID: 0000-0002-3254-5238 AD - Department of Oral Morphology, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Okayama 700-8525, Japan. FAU - Kozaki, Ken-Ichi AU - Kozaki KI AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. FAU - Calderwood, Stuart K AU - Calderwood SK AD - Division of Molecular and Cellular Biology, Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA. FAU - Takigawa, Masaharu AU - Takigawa M AD - Advanced Research Center for Oral and Craniofacial Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. FAU - Okamoto, Kuniaki AU - Okamoto K AD - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan. LA - eng GR - JP17K17895/Japan Society for the Promotion of Science/ GR - JP17K11642/Japan Society for the Promotion of Science/ GR - JP19H03817/Japan Society for the Promotion of Science/ GR - 19H04051/Japan Society for the Promotion of Science/ GR - JP17K11643/Japan Society for the Promotion of Science/ GR - 17K11669/Japan Society for the Promotion of Science/ GR - 18K09789/Japan Society for the Promotion of Science/ GR - TE/Suzuken Memorial Foundation/ GR - KOk, CS, TE/Ryobi Teien Memory Foundation/ PT - Journal Article DEP - 20200404 PL - Switzerland TA - Cancers (Basel) JT - Cancers JID - 101526829 PMC - PMC7226423 OTO - NOTNLM OT - CCN2/CTGF OT - CRISPR OT - cellular communication network factor OT - extracellular vesicles OT - genome editing OT - matrix metalloproteinase OT - moonlighting metalloproteinase (MMP) OT - oncosome OT - protein moonlighting OT - transcription factor COIS- The authors have no competing financial interests to declare. EDAT- 2020/04/09 06:00 MHDA- 2020/04/09 06:01 PMCR- 2020/04/04 CRDT- 2020/04/09 06:00 PHST- 2020/02/19 00:00 [received] PHST- 2020/03/24 00:00 [revised] PHST- 2020/04/02 00:00 [accepted] PHST- 2020/04/09 06:00 [entrez] PHST- 2020/04/09 06:00 [pubmed] PHST- 2020/04/09 06:01 [medline] PHST- 2020/04/04 00:00 [pmc-release] AID - cancers12040881 [pii] AID - cancers-12-00881 [pii] AID - 10.3390/cancers12040881 [doi] PST - epublish SO - Cancers (Basel). 2020 Apr 4;12(4):881. doi: 10.3390/cancers12040881.