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Drug target mining and analysis of the Chinese tree shrew for pharmacological testing.

Zhao F, Guo X, Wang Y, Liu J, Lee WH, Zhang Y - PLoS ONE (2014)

Bottom Line: The discovery of new drugs requires the development of improved animal models for drug testing.The Chinese tree shrew is considered to be a realistic candidate model.Target validation also demonstrated that the constitutive expression of the proteinase-activated receptors of tree shrew platelets is similar to that of human platelets but differs from that of mouse platelets.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, PR China; Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming, Yunnan, PR China.

ABSTRACT
The discovery of new drugs requires the development of improved animal models for drug testing. The Chinese tree shrew is considered to be a realistic candidate model. To assess the potential of the Chinese tree shrew for pharmacological testing, we performed drug target prediction and analysis on genomic and transcriptomic scales. Using our pipeline, 3,482 proteins were predicted to be drug targets. Of these predicted targets, 446 and 1,049 proteins with the highest rank and total scores, respectively, included homologs of targets for cancer chemotherapy, depression, age-related decline and cardiovascular disease. Based on comparative analyses, more than half of drug target proteins identified from the tree shrew genome were shown to be higher similarity to human targets than in the mouse. Target validation also demonstrated that the constitutive expression of the proteinase-activated receptors of tree shrew platelets is similar to that of human platelets but differs from that of mouse platelets. We developed an effective pipeline and search strategy for drug target prediction and the evaluation of model-based target identification for drug testing. This work provides useful information for future studies of the Chinese tree shrew as a source of novel targets for drug discovery research.

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Related in: MedlinePlus

Protease-activated receptors (PARs) present on tree shrew platelets.(A) Amplification of PARs from different tissues using RT-PCR. Primers were designed based on the original tree shrew sequences; β-actin was used as a control. (B) Detection of PAR expression in different tissues by western blotting. PAR anti-human antibodies were used. (C) Surface expression of PAR1 and PAR4 in tree shrew platelets analyzed by flow cytometry. (D) Tree shrew PAR expression measured by qRT-PCR and flow cytometry. The qRT-PCR results are presented as the mean ± SEM (three technical replicates) using β-actin as an internal control.
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pone-0104191-g005: Protease-activated receptors (PARs) present on tree shrew platelets.(A) Amplification of PARs from different tissues using RT-PCR. Primers were designed based on the original tree shrew sequences; β-actin was used as a control. (B) Detection of PAR expression in different tissues by western blotting. PAR anti-human antibodies were used. (C) Surface expression of PAR1 and PAR4 in tree shrew platelets analyzed by flow cytometry. (D) Tree shrew PAR expression measured by qRT-PCR and flow cytometry. The qRT-PCR results are presented as the mean ± SEM (three technical replicates) using β-actin as an internal control.

Mentions: To illustrate the usefulness and potential of our large-scale approach, we validated the activation of a predicted target using proteinase-activated receptors (PARs), which are members of a GPCR family. The roles of these receptors in cancer as well as in cardiovascular, musculoskeletal, gastrointestinal, respiratory and central nervous system disorders have recently been discovered. PAR expression was detected in the tree shrew, in tree shrews platelets, PAR1 and PAR4 are exclusively detected (Figures 5A and 5D). Similar results were obtained by western blotting (Figure 5B) and flow cytometry analysis (Figure 5C). Furthermore, PAR1, 2, 3 and 4 were found to be highly expressed in tree shrew brain, muscle, stomach and small intestine tissues, at least at the mRNA levels (Figure 5D). These results indicate that the constitutive expression of PAR is similar in tree shrew and human platelets [14] and distinct from that observed in mouse platelets, which express PAR3 and PAR4 [15].


Drug target mining and analysis of the Chinese tree shrew for pharmacological testing.

Zhao F, Guo X, Wang Y, Liu J, Lee WH, Zhang Y - PLoS ONE (2014)

Protease-activated receptors (PARs) present on tree shrew platelets.(A) Amplification of PARs from different tissues using RT-PCR. Primers were designed based on the original tree shrew sequences; β-actin was used as a control. (B) Detection of PAR expression in different tissues by western blotting. PAR anti-human antibodies were used. (C) Surface expression of PAR1 and PAR4 in tree shrew platelets analyzed by flow cytometry. (D) Tree shrew PAR expression measured by qRT-PCR and flow cytometry. The qRT-PCR results are presented as the mean ± SEM (three technical replicates) using β-actin as an internal control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126716&req=5

pone-0104191-g005: Protease-activated receptors (PARs) present on tree shrew platelets.(A) Amplification of PARs from different tissues using RT-PCR. Primers were designed based on the original tree shrew sequences; β-actin was used as a control. (B) Detection of PAR expression in different tissues by western blotting. PAR anti-human antibodies were used. (C) Surface expression of PAR1 and PAR4 in tree shrew platelets analyzed by flow cytometry. (D) Tree shrew PAR expression measured by qRT-PCR and flow cytometry. The qRT-PCR results are presented as the mean ± SEM (three technical replicates) using β-actin as an internal control.
Mentions: To illustrate the usefulness and potential of our large-scale approach, we validated the activation of a predicted target using proteinase-activated receptors (PARs), which are members of a GPCR family. The roles of these receptors in cancer as well as in cardiovascular, musculoskeletal, gastrointestinal, respiratory and central nervous system disorders have recently been discovered. PAR expression was detected in the tree shrew, in tree shrews platelets, PAR1 and PAR4 are exclusively detected (Figures 5A and 5D). Similar results were obtained by western blotting (Figure 5B) and flow cytometry analysis (Figure 5C). Furthermore, PAR1, 2, 3 and 4 were found to be highly expressed in tree shrew brain, muscle, stomach and small intestine tissues, at least at the mRNA levels (Figure 5D). These results indicate that the constitutive expression of PAR is similar in tree shrew and human platelets [14] and distinct from that observed in mouse platelets, which express PAR3 and PAR4 [15].

Bottom Line: The discovery of new drugs requires the development of improved animal models for drug testing.The Chinese tree shrew is considered to be a realistic candidate model.Target validation also demonstrated that the constitutive expression of the proteinase-activated receptors of tree shrew platelets is similar to that of human platelets but differs from that of mouse platelets.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, PR China; Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming, Yunnan, PR China.

ABSTRACT
The discovery of new drugs requires the development of improved animal models for drug testing. The Chinese tree shrew is considered to be a realistic candidate model. To assess the potential of the Chinese tree shrew for pharmacological testing, we performed drug target prediction and analysis on genomic and transcriptomic scales. Using our pipeline, 3,482 proteins were predicted to be drug targets. Of these predicted targets, 446 and 1,049 proteins with the highest rank and total scores, respectively, included homologs of targets for cancer chemotherapy, depression, age-related decline and cardiovascular disease. Based on comparative analyses, more than half of drug target proteins identified from the tree shrew genome were shown to be higher similarity to human targets than in the mouse. Target validation also demonstrated that the constitutive expression of the proteinase-activated receptors of tree shrew platelets is similar to that of human platelets but differs from that of mouse platelets. We developed an effective pipeline and search strategy for drug target prediction and the evaluation of model-based target identification for drug testing. This work provides useful information for future studies of the Chinese tree shrew as a source of novel targets for drug discovery research.

Show MeSH
Related in: MedlinePlus