Exosomal miRNA confers chemo resistance via targeting Cav1/p-gp/M2-type macrophage axis in ovarian cancer

Pinar Kanlikilicer; Recep Bayraktar; Merve Denizli; Mohammed H Rashed; Cristina Ivan; Burcu Aslan; Rahul Mitra; Kubra Karagoz; Emine Bayraktar; Xinna Zhang; Cristian Rodriguez-Aguayo; Amr Ahmed El-Arabey; Nermin Kahraman; Seyda Baydogan; Ozgur Ozkayar; Michael L Gatza; Bulent Ozpolat; George A Calin; Anil K Sood; Gabriel Lopez-Berestein

doi:10.1016/j.ebiom.2018.11.004

Exosomal miRNA confers chemo resistance via targeting Cav1/p-gp/M2-type macrophage axis in ovarian cancer

EBioMedicine. 2018 Dec:38:100-112. doi: 10.1016/j.ebiom.2018.11.004. Epub 2018 Nov 25.

Authors

Pinar Kanlikilicer¹, Recep Bayraktar², Merve Denizli², Mohammed H Rashed², Cristina Ivan³, Burcu Aslan¹, Rahul Mitra⁴, Kubra Karagoz⁵, Emine Bayraktar², Xinna Zhang³, Cristian Rodriguez-Aguayo¹, Amr Ahmed El-Arabey², Nermin Kahraman², Seyda Baydogan², Ozgur Ozkayar⁶, Michael L Gatza⁵, Bulent Ozpolat¹, George A Calin⁷, Anil K Sood⁸, Gabriel Lopez-Berestein⁹

Affiliations

¹ Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
² Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
³ Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Department of Gynecologic Oncology and Reproductive Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
⁴ Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
⁵ Rutgers Cancer Institute of New Jersey, New Brunswick, New Jersey, USA.
⁶ Hacettepe University, Ankara, Turkey.
⁷ Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
⁸ Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Department of Gynecologic Oncology and Reproductive Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
⁹ Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA; Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA. Electronic address: glopez@mdanderson.org.

Abstract

Background: Circulating miRNAs are known to play important roles in intercellular communication. However, the effects of exosomal miRNAs on cells are not fully understood.

Methods: To investigate the role of exosomal miR-1246 in ovarian cancer (OC) microenvironment, we performed RPPA as well as many other in vitro functional assays in ovarian cancer cells (sensitive; HeyA8, Skov3ip1, A2780 and chemoresistant; HeyA8-MDR, Skov3-TR, A2780-CP20). Therapeutic effect of miR-1246 inhibitor treatment was tested in OC animal model. We showed the effect of OC exosomal miR-1246 uptake on macrophages by co-culture experiments.

Findings: Substantial expression of oncogenic miR-1246 OC exosomes was found. We showed that Cav1 gene, which is the direct target of miR-1246, is involved in the process of exosomal transfer. A significantly worse overall prognosis were found for OC patients with high miR-1246 and low Cav1 expression based on TCGA data. miR-1246 expression were significantly higher in paclitaxel-resistant OC exosomes than in their sensitive counterparts. Overexpression of Cav1 and anti-miR-1246 treatment significantly sensitized OC cells to paclitaxel. We showed that Cav1 and multi drug resistance (MDR) gene is involved in the process of exosomal transfer. Our proteomic approach also revealed that miR-1246 inhibits Cav1 and acts through PDGFβ receptor at the recipient cells to inhibit cell proliferation. miR-1246 inhibitor treatment in combination with chemotherapy led to reduced tumor burden in vivo. Finally, we demonstrated that when OC cells are co-cultured with macrophages, they are capable of transferring their oncogenic miR-1246 to M2-type macrophages, but not M0-type macrophages.

Interpretation: Our results suggest that cancer exosomes may contribute to oncogenesis by manipulating neighboring infiltrating immune cells. This study provide a new mechanistic therapeutic approach to overcome chemoresistance and tumor progression through exosomal miR-1246 in OC patients.

Keywords: Cav1; Exosome; Ovarian cancer; P-gp; miR-1246; oncomiR.

MeSH terms

ATP Binding Cassette Transporter, Subfamily B / genetics*
Animals
Apoptosis / drug effects
Caveolin 1 / genetics*
Caveolin 1 / metabolism
Cell Line, Tumor
Cell Proliferation
Disease Models, Animal
Drug Resistance, Neoplasm / genetics*
Exosomes* / metabolism
Female
Gene Expression Profiling
Gene Expression Regulation, Neoplastic
Humans
Macrophages / drug effects*
Macrophages / metabolism*
Mice
MicroRNAs / genetics*
MicroRNAs / metabolism
Models, Biological
Ovarian Neoplasms / drug therapy
Ovarian Neoplasms / genetics*
Ovarian Neoplasms / metabolism
Ovarian Neoplasms / mortality
RNA Interference
Receptor, Platelet-Derived Growth Factor beta / metabolism
Signal Transduction
Tumor Microenvironment

Substances

ATP Binding Cassette Transporter, Subfamily B
CAV1 protein, human
Caveolin 1
MicroRNAs
Receptor, Platelet-Derived Growth Factor beta

Abstract

MeSH terms

Substances

Grants and funding