Targeting ABCB1 (MDR1) in multi-drug resistant osteosarcoma cells using the CRISPR-Cas9 system to reverse drug resistance.
BACKGROUND
Multi-drug resistance (MDR) remains a significant obstacle to successful chemotherapy treatment for osteosarcoma patients. One of the central causes of MDR is the overexpression of the membrane bound drug transporter protein P-glycoprotein (P-gp), which is the protein product of the MDR gene ABCB1. Though several methods have been reported to reverse MDR in vitro and in vivo when combined with anticancer drugs, they have yet to be proven useful in the clinical setting.


RESULTS
The meta-analysis demonstrated that a high level of P-gp may predict poor survival in patients with osteosarcoma. The expression of P-gp can be efficiently blocked by the clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9 system (CRISPR-Cas9). Inhibition of ABCB1 was associated with reversing drug resistance in osteosarcoma MDR cell lines (KHOSR2 and U-2OSR2) to doxorubicin.


METHODS
We performed a meta-analysis to investigate the relationship between P-gp expression and survival in patients with osteosarcoma. Then we adopted the CRISPR-Cas9, a robust and highly efficient novel genome editing tool, to determine its effect on reversing drug resistance by targeting endogenous ABCB1 gene at the DNA level in osteosarcoma MDR cell lines.


CONCLUSIONS
These results suggest that the CRISPR-Cas9 system is a useful tool for the modification of ABCB1 gene, and may be useful in extending the long-term efficacy of chemotherapy by overcoming P-gp-mediated MDR in the clinical setting.
Department of Orthopaedics, The 2nd Xiangya Hospital of Central South University, Changsha, Hunan, 410011, P.R. China.|Sarcoma Biology Laboratory, Department of Orthopaedic surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
Tang
T
Liu
Department of Orthopaedics, The 2nd Xiangya Hospital of Central South University, Changsha, Hunan, 410011, P.R. China.
Zhihong
Z
Li
Department of Orthopaedics, The 2nd Xiangya Hospital of Central South University, Changsha, Hunan, 410011, P.R. China.
Qing
Q
Zhang
Sarcoma Biology Laboratory, Department of Orthopaedic surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
Karen
K
De Amorim Bernstein
Sarcoma Biology Laboratory, Department of Orthopaedic surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
Santiago
S
Lozano-Calderon
Sarcoma Biology Laboratory, Department of Orthopaedic surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
Edwin
E
Choy
Sarcoma Biology Laboratory, Department of Orthopaedic surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
Francis J
FJ
Hornicek
Sarcoma Biology Laboratory, Department of Orthopaedic surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
Zhenfeng
Z
Duan
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology*; D018432:Drug Resistance, Multiple / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D012516:Osteosarcoma / Q000188:drug therapy* / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D014162:Transfection
C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D000903:Antibiotics, Antineoplastic; D004317:Doxorubicin
ABCB1; CRISPR-Cas9; P-glycoprotein; meta-analysis; osteosarcoma