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Integrated Genomics of Crohn's Disease Risk Variant Identifies a Role for CLEC12A in Antibacterial Autophagy.

Begun J, Lassen KG, Jijon HB, Baxt LA, Goel G, Heath RJ, Ng A, Tam JM, Kuo SY, Villablanca EJ, Fagbami L, Oosting M, Kumar V, Schenone M, Carr SA, Joosten LA, Vyas JM, Daly MJ, Netea MG, Brown GD, Wijmenga C, Xavier RJ - Cell Rep (2015)

Bottom Line: Antibacterial autophagy is impaired in CLEC12A-deficient cells, and this effect is exacerbated in the presence of the ATG16L1(∗)300A risk allele.Integrated genomics identified a functional interaction between CLEC12A and an E3-ubiquitin ligase complex that functions in antibacterial autophagy.These data identify CLEC12A as early adaptor molecule for antibacterial autophagy and highlight perturbational profiling as a method to elucidate defense pathways in complex genetic disease.

View Article: PubMed Central - PubMed

Affiliation: Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Boston, MA 02114, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Mater Research Institute, University of Queensland, Brisbane, QLD 4101, Australia.

No MeSH data available.


Related in: MedlinePlus

Clec12a−/− Mice Are More Susceptible to Salmonella Infection In Vivo(A) Confocal micrographs of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs at 1 hr post-infection. The scale bars represent 10 μm.(B) Quantification of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs. Data shown represent mean ± SD of n = 3 independent experiments; unpaired t test. ∗∗p < 0.01.(C and D) Salmonella cfus quantified per gram of stool (C) and per organ (D) at 4 days post-infection are displayed. Data are shown as mean ± SD; ∗∗p ≤ 0.01. Data are representative of at least two independent experiments (n = 9 for WT; n = 7 or 8 for Clec12a−/−).(E) Clinical score for infected mice at 4 days post-infection. Data are shown as mean ± SD. Data are representative of at least two independent experiments. ∗∗∗∗p < 0.0001; unpaired t test. (n = 14 for WT; n = 13 for Clec12a−/−).(F) Survival curve for infected WT and Clec12a−/− mice (n = 9 mice per genotype).See also Figure S3.
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fig3: Clec12a−/− Mice Are More Susceptible to Salmonella Infection In Vivo(A) Confocal micrographs of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs at 1 hr post-infection. The scale bars represent 10 μm.(B) Quantification of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs. Data shown represent mean ± SD of n = 3 independent experiments; unpaired t test. ∗∗p < 0.01.(C and D) Salmonella cfus quantified per gram of stool (C) and per organ (D) at 4 days post-infection are displayed. Data are shown as mean ± SD; ∗∗p ≤ 0.01. Data are representative of at least two independent experiments (n = 9 for WT; n = 7 or 8 for Clec12a−/−).(E) Clinical score for infected mice at 4 days post-infection. Data are shown as mean ± SD. Data are representative of at least two independent experiments. ∗∗∗∗p < 0.0001; unpaired t test. (n = 14 for WT; n = 13 for Clec12a−/−).(F) Survival curve for infected WT and Clec12a−/− mice (n = 9 mice per genotype).See also Figure S3.

Mentions: To determine whether CLEC12A plays a role in pathogen defense in vivo, we used Clec12a−/− mice. Bone-marrow-derived macrophages (BMDMs) from WT or Clec12a−/− mice were infected with a strain of Listeria (EGDe) known to be susceptible to autophagy in primary macrophages (Anand et al., 2011; Birmingham et al., 2007; Huett et al., 2012). Consistent with the Salmonella infection results in siRNA-treated cells, Clec12a−/− BMDMs displayed lower levels of bacterial colocalization with LC3 compared with WT BMDMs (Figures 3A and 3B). Additionally, Clec12a−/− BMDMs did not show differences in Torin-1-induced autophagy, suggesting that bulk autophagy is normal in these cells (Figure S3B). Taken together, these data suggest that CLEC12A functions selectively in the antibacterial autophagy pathway for multiple pathogens in both mouse and human cells.


Integrated Genomics of Crohn's Disease Risk Variant Identifies a Role for CLEC12A in Antibacterial Autophagy.

Begun J, Lassen KG, Jijon HB, Baxt LA, Goel G, Heath RJ, Ng A, Tam JM, Kuo SY, Villablanca EJ, Fagbami L, Oosting M, Kumar V, Schenone M, Carr SA, Joosten LA, Vyas JM, Daly MJ, Netea MG, Brown GD, Wijmenga C, Xavier RJ - Cell Rep (2015)

Clec12a−/− Mice Are More Susceptible to Salmonella Infection In Vivo(A) Confocal micrographs of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs at 1 hr post-infection. The scale bars represent 10 μm.(B) Quantification of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs. Data shown represent mean ± SD of n = 3 independent experiments; unpaired t test. ∗∗p < 0.01.(C and D) Salmonella cfus quantified per gram of stool (C) and per organ (D) at 4 days post-infection are displayed. Data are shown as mean ± SD; ∗∗p ≤ 0.01. Data are representative of at least two independent experiments (n = 9 for WT; n = 7 or 8 for Clec12a−/−).(E) Clinical score for infected mice at 4 days post-infection. Data are shown as mean ± SD. Data are representative of at least two independent experiments. ∗∗∗∗p < 0.0001; unpaired t test. (n = 14 for WT; n = 13 for Clec12a−/−).(F) Survival curve for infected WT and Clec12a−/− mice (n = 9 mice per genotype).See also Figure S3.
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fig3: Clec12a−/− Mice Are More Susceptible to Salmonella Infection In Vivo(A) Confocal micrographs of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs at 1 hr post-infection. The scale bars represent 10 μm.(B) Quantification of LC3-Listeria colocalization in WT and Clec12a−/− BMDMs. Data shown represent mean ± SD of n = 3 independent experiments; unpaired t test. ∗∗p < 0.01.(C and D) Salmonella cfus quantified per gram of stool (C) and per organ (D) at 4 days post-infection are displayed. Data are shown as mean ± SD; ∗∗p ≤ 0.01. Data are representative of at least two independent experiments (n = 9 for WT; n = 7 or 8 for Clec12a−/−).(E) Clinical score for infected mice at 4 days post-infection. Data are shown as mean ± SD. Data are representative of at least two independent experiments. ∗∗∗∗p < 0.0001; unpaired t test. (n = 14 for WT; n = 13 for Clec12a−/−).(F) Survival curve for infected WT and Clec12a−/− mice (n = 9 mice per genotype).See also Figure S3.
Mentions: To determine whether CLEC12A plays a role in pathogen defense in vivo, we used Clec12a−/− mice. Bone-marrow-derived macrophages (BMDMs) from WT or Clec12a−/− mice were infected with a strain of Listeria (EGDe) known to be susceptible to autophagy in primary macrophages (Anand et al., 2011; Birmingham et al., 2007; Huett et al., 2012). Consistent with the Salmonella infection results in siRNA-treated cells, Clec12a−/− BMDMs displayed lower levels of bacterial colocalization with LC3 compared with WT BMDMs (Figures 3A and 3B). Additionally, Clec12a−/− BMDMs did not show differences in Torin-1-induced autophagy, suggesting that bulk autophagy is normal in these cells (Figure S3B). Taken together, these data suggest that CLEC12A functions selectively in the antibacterial autophagy pathway for multiple pathogens in both mouse and human cells.

Bottom Line: Antibacterial autophagy is impaired in CLEC12A-deficient cells, and this effect is exacerbated in the presence of the ATG16L1(∗)300A risk allele.Integrated genomics identified a functional interaction between CLEC12A and an E3-ubiquitin ligase complex that functions in antibacterial autophagy.These data identify CLEC12A as early adaptor molecule for antibacterial autophagy and highlight perturbational profiling as a method to elucidate defense pathways in complex genetic disease.

View Article: PubMed Central - PubMed

Affiliation: Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Boston, MA 02114, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Mater Research Institute, University of Queensland, Brisbane, QLD 4101, Australia.

No MeSH data available.


Related in: MedlinePlus