Limits...
Molecular insight into the association between cartilage regeneration and ear wound healing in genetic mouse models: targeting new genes in regeneration.

Rai MF, Schmidt EJ, McAlinden A, Cheverud JM, Sandell LJ - G3 (Bethesda) (2013)

Bottom Line: The expression of several candidates was significantly heritable among strains.Cluster analysis accurately classified healers and nonhealers for seven out of eight strains based on gene expression.Specific sequence differences between LG/J and SM/J were identified as potential causal polymorphisms.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Washington University School of Medicine at Barnes-Jewish Hospital, St. Louis, Missouri 63110.

ABSTRACT
Tissue regeneration is a complex trait with few genetic models available. Mouse strains LG/J and MRL are exceptional healers. Using recombinant inbred strains from a large (LG/J, healer) and small (SM/J, nonhealer) intercross, we have previously shown a positive genetic correlation between ear wound healing, knee cartilage regeneration, and protection from osteoarthritis. We hypothesize that a common set of genes operates in tissue healing and articular cartilage regeneration. Taking advantage of archived histological sections from recombinant inbred strains, we analyzed expression of candidate genes through branched-chain DNA technology directly from tissue lysates. We determined broad-sense heritability of candidates, Pearson correlation of candidates with healing phenotypes, and Ward minimum variance cluster analysis for strains. A bioinformatic assessment of allelic polymorphisms within and near candidate genes was also performed. The expression of several candidates was significantly heritable among strains. Although several genes correlated with both ear wound healing and cartilage healing at a marginal level, the expression of four genes representing DNA repair (Xrcc2, Pcna) and Wnt signaling (Axin2, Wnt16) pathways was significantly positively correlated with both phenotypes. Cluster analysis accurately classified healers and nonhealers for seven out of eight strains based on gene expression. Specific sequence differences between LG/J and SM/J were identified as potential causal polymorphisms. Our study suggests a common genetic basis between tissue healing and osteoarthritis susceptibility. Mapping genetic variations causing differences in diverse healing responses in multiple tissues may reveal generic healing processes in pursuit of new therapeutic targets designed to induce or enhance regeneration and, potentially, protection from osteoarthritis.

Show MeSH

Related in: MedlinePlus

Cluster analysis of normalized gene expression levels. Knee joint tissues from FFPE sections were macrodissected and tissue homogenates were prepared to quantify candidate genes through QuantiGene Plex assay. The gene expression levels were used to calculate fold difference among strains compared to C57BL/6J (a nonhealer control strain) as shown in a seven-tiered categorical scale. Hierarchical clustering analysis was performed using Ward minimum variance method to produce a dendrogram. As shown here, the healer strains (except for LGXSM-33) clustered together exclusive of other nonhealers and intermediate healers, indicating an association between gene expression levels and healing potential.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3815053&req=5

fig3: Cluster analysis of normalized gene expression levels. Knee joint tissues from FFPE sections were macrodissected and tissue homogenates were prepared to quantify candidate genes through QuantiGene Plex assay. The gene expression levels were used to calculate fold difference among strains compared to C57BL/6J (a nonhealer control strain) as shown in a seven-tiered categorical scale. Hierarchical clustering analysis was performed using Ward minimum variance method to produce a dendrogram. As shown here, the healer strains (except for LGXSM-33) clustered together exclusive of other nonhealers and intermediate healers, indicating an association between gene expression levels and healing potential.

Mentions: The most interesting finding from the cluster analysis was the classification of strains into two distinct clusters largely (but not exclusively) corresponding to their healing ability (Rai et al. 2012) with 100% bootstrap support (1000 iterations) (Figure 3). One cluster represented nonhealers and moderate healers, including SM/J, C57BL/6J, LGXSM-5, and LGXSM-35, whereas the other cluster represented predominately classical healers LG/J, LGXSM-6, and MRL/MpJ. Surprisingly, strain LGXSM-33, a poor healer, was clustered with MRL/MpJ, a super-healer.


Molecular insight into the association between cartilage regeneration and ear wound healing in genetic mouse models: targeting new genes in regeneration.

Rai MF, Schmidt EJ, McAlinden A, Cheverud JM, Sandell LJ - G3 (Bethesda) (2013)

Cluster analysis of normalized gene expression levels. Knee joint tissues from FFPE sections were macrodissected and tissue homogenates were prepared to quantify candidate genes through QuantiGene Plex assay. The gene expression levels were used to calculate fold difference among strains compared to C57BL/6J (a nonhealer control strain) as shown in a seven-tiered categorical scale. Hierarchical clustering analysis was performed using Ward minimum variance method to produce a dendrogram. As shown here, the healer strains (except for LGXSM-33) clustered together exclusive of other nonhealers and intermediate healers, indicating an association between gene expression levels and healing potential.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3815053&req=5

fig3: Cluster analysis of normalized gene expression levels. Knee joint tissues from FFPE sections were macrodissected and tissue homogenates were prepared to quantify candidate genes through QuantiGene Plex assay. The gene expression levels were used to calculate fold difference among strains compared to C57BL/6J (a nonhealer control strain) as shown in a seven-tiered categorical scale. Hierarchical clustering analysis was performed using Ward minimum variance method to produce a dendrogram. As shown here, the healer strains (except for LGXSM-33) clustered together exclusive of other nonhealers and intermediate healers, indicating an association between gene expression levels and healing potential.
Mentions: The most interesting finding from the cluster analysis was the classification of strains into two distinct clusters largely (but not exclusively) corresponding to their healing ability (Rai et al. 2012) with 100% bootstrap support (1000 iterations) (Figure 3). One cluster represented nonhealers and moderate healers, including SM/J, C57BL/6J, LGXSM-5, and LGXSM-35, whereas the other cluster represented predominately classical healers LG/J, LGXSM-6, and MRL/MpJ. Surprisingly, strain LGXSM-33, a poor healer, was clustered with MRL/MpJ, a super-healer.

Bottom Line: The expression of several candidates was significantly heritable among strains.Cluster analysis accurately classified healers and nonhealers for seven out of eight strains based on gene expression.Specific sequence differences between LG/J and SM/J were identified as potential causal polymorphisms.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Washington University School of Medicine at Barnes-Jewish Hospital, St. Louis, Missouri 63110.

ABSTRACT
Tissue regeneration is a complex trait with few genetic models available. Mouse strains LG/J and MRL are exceptional healers. Using recombinant inbred strains from a large (LG/J, healer) and small (SM/J, nonhealer) intercross, we have previously shown a positive genetic correlation between ear wound healing, knee cartilage regeneration, and protection from osteoarthritis. We hypothesize that a common set of genes operates in tissue healing and articular cartilage regeneration. Taking advantage of archived histological sections from recombinant inbred strains, we analyzed expression of candidate genes through branched-chain DNA technology directly from tissue lysates. We determined broad-sense heritability of candidates, Pearson correlation of candidates with healing phenotypes, and Ward minimum variance cluster analysis for strains. A bioinformatic assessment of allelic polymorphisms within and near candidate genes was also performed. The expression of several candidates was significantly heritable among strains. Although several genes correlated with both ear wound healing and cartilage healing at a marginal level, the expression of four genes representing DNA repair (Xrcc2, Pcna) and Wnt signaling (Axin2, Wnt16) pathways was significantly positively correlated with both phenotypes. Cluster analysis accurately classified healers and nonhealers for seven out of eight strains based on gene expression. Specific sequence differences between LG/J and SM/J were identified as potential causal polymorphisms. Our study suggests a common genetic basis between tissue healing and osteoarthritis susceptibility. Mapping genetic variations causing differences in diverse healing responses in multiple tissues may reveal generic healing processes in pursuit of new therapeutic targets designed to induce or enhance regeneration and, potentially, protection from osteoarthritis.

Show MeSH
Related in: MedlinePlus