Silencing AXL by covalent siRNA-gelatin-antibody nanoconjugate inactivates mTOR/EMT pathway and stimulates p53 for TKI sensitization in NSCLC

Dhananjay Suresh; Ajit Zambre; Soumavo Mukherjee; Shreya Ghoshdastidar; Yuexu Jiang; Trupti Joshi; Anandhi Upendran; Raghuraman Kannan

doi:10.1016/j.nano.2019.04.010

Silencing AXL by covalent siRNA-gelatin-antibody nanoconjugate inactivates mTOR/EMT pathway and stimulates p53 for TKI sensitization in NSCLC

Nanomedicine. 2019 Aug:20:102007. doi: 10.1016/j.nano.2019.04.010. Epub 2019 May 12.

Authors

Dhananjay Suresh¹, Ajit Zambre², Soumavo Mukherjee¹, Shreya Ghoshdastidar¹, Yuexu Jiang³, Trupti Joshi⁴, Anandhi Upendran⁵, Raghuraman Kannan⁶

Affiliations

¹ Department of Bioengineering, University of Missouri, Columbia, MO, USA.
² Department of Radiology, University of Missouri, Columbia, MO, USA.
³ Department of Electrical Engineering and Computer Science, University of Missouri, Columbia, MO, USA.
⁴ Department of Health Management and Informatics, University of Missouri, Columbia, MO, USA; Department of MU Informatics Institute, University of Missouri, Columbia, MO, USA.
⁵ Department of Medical Pharmacology and Physiology, University of Missouri, Columbia, MO, USA; MU-Institute of Clinical and Translational Science (MU-iCATS), University of Missouri, Columbia, MO, USA.
⁶ Department of Bioengineering, University of Missouri, Columbia, MO, USA; Department of Radiology, University of Missouri, Columbia, MO, USA. Electronic address: kannanr@health.missouri.edu.

PMID: 31085346
DOI: 10.1016/j.nano.2019.04.010

Abstract

Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality with the 5-year survival rate at a dismal 16% for the past 40 years. Drug resistance is a major obstacle to achieving long-term patient survival. Identifying and validating molecular biomarkers responsible for resistance and thereby adopting multi-directional therapy is necessary to improve the survival rate. Previous studies indicated ~20% of tyrosine kinase inhibitor (TKI) resistant NSCLC patients overexpress AXL with increase in EMT and decrease in p53 expression. To overcome the resistance, we designed gelatin nanoparticles covalently conjugated with EGFR targeting antibody and siRNA (GAbsiAXL). GAbsiAXL efficiently silences AXL, decreases mTOR and EMT signaling with concomitant increase in p53 expression. Because of the molecular changes, the AXL silencing sensitizes the cells to TKI. Our results show AXL overexpression has an important role in driving TKI resistance through close association with energy-dependent mitochondrial pathways.

Keywords: AXL; Gelatin NP; NSCLC; TKI; siRNA.

MeSH terms

Antibodies / chemistry
Axl Receptor Tyrosine Kinase
Carcinoma, Non-Small-Cell Lung / drug therapy*
Cell Line, Tumor
Down-Regulation / drug effects
Drug Resistance, Neoplasm / drug effects
Epithelial-Mesenchymal Transition* / drug effects
Gelatin / chemistry
Gene Regulatory Networks
Gene Silencing
Humans
Lung Neoplasms
Matrix Metalloproteinases / metabolism
Nanoconjugates / chemistry*
Phosphatidylinositol 3-Kinases / metabolism
Protein Kinase Inhibitors / pharmacology
Protein Kinase Inhibitors / therapeutic use*
Proto-Oncogene Proteins / metabolism*
Proto-Oncogene Proteins c-akt / metabolism
RNA, Small Interfering / metabolism*
Receptor Protein-Tyrosine Kinases / metabolism*
TOR Serine-Threonine Kinases / metabolism*
Tumor Suppressor Protein p53 / metabolism*

Substances

Antibodies
Nanoconjugates
Protein Kinase Inhibitors
Proto-Oncogene Proteins
RNA, Small Interfering
Tumor Suppressor Protein p53
Gelatin
Receptor Protein-Tyrosine Kinases
Proto-Oncogene Proteins c-akt
TOR Serine-Threonine Kinases
Matrix Metalloproteinases
Axl Receptor Tyrosine Kinase
AXL protein, human