Identification of novel host-oriented targets for Human Immunodeficiency Virus type 1 using Random Homozygous Gene Perturbation.
BACKGROUND
Human Immunodeficiency Virus (HIV) is a global threat to public health. Current therapies that directly target the virus often are rendered ineffective due to the emergence of drug-resistant viral variants. An emerging concept to combat drug resistance is the idea of targeting host mechanisms that are essential for the propagation of the virus, but have a minimal cellular effect.


RESULTS
Herein, using Random Homozygous Gene Perturbation (RHGP), we have identified cellular targets that allow human MT4 cells to survive otherwise lethal infection by a wild type HIV-1NL4-3. These gene targets were validated by the reversibility of the RHGP technology, which confirmed that the RHGP itself was responsible for the resistance to HIV-1 infection. We further confirmed by siRNA knockdowns that the RHGP-identified cellular pathways are responsible for resistance to infection by either CXCR4 or CCR5 tropic HIV-1 variants. We also demonstrated that cell clones with these gene targets disrupted by RHGP were not permissible to the replication of a drug resistant HIV-1 mutant.


CONCLUSIONS
These studies demonstrate the power of RHGP to identify novel host targets that are essential for the viral life cycle but which can be safely perturbed without overt cytotoxicity. These findings suggest opportunities for the future development of host-oriented therapeutics with the broad spectrum potential for safe and effective inhibition of HIV infection.
Functional Genetics, Inc, Gaithersburg, MD 20878, USA. hmao@functional-genetics.com
Hanwen
H
Mao
Hanson
H
Chen
Zena
Z
Fesseha
Shaojing
S
Chang
Huong
H
Ung-Medoff
Jessica
J
Van Dyke
Manu
M
Kohli
Wu-Bo
WB
Li
Michael
M
Goldblatt
Michael S
MS
Kinch
D002460:Cell Line; D002470:Cell Survival; D055785:Gene Knockdown Techniques; D015497:HIV-1 / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication
D034741:RNA, Small Interfering