Requirements for Acute Burn and Chronic Surgical Wound Infection
Soft tissue infections, such as those in burns, bed sores, and diabetic ulcers, are a significant healthcare and economic burden in the developed and developing world. The opportunistic pathogen P. aeruginosa can cause both acute and chronic infections, and the trajectory of these two types of infections is vastly different. We used high-throughput sequencing to profile P. aeruginosa genome-wide gene expression and mutant fitness during mouse model acute and non-diabetic chronic wound infections. Using these data, we show that wounds are nutrient-rich growth environments in which long-chain fatty acids are a primary source of carbon and energy. We also show that the ability to travel along spatio-chemical gradients by chemotaxis is critical for bacterial fitness and virulence in acute but not chronic infections. Our results demonstrate the utility of simultaneous mutant fitness and gene expression profiling to discover critical functions in complex growth environments.
Vyšlo v časopise:
Requirements for Acute Burn and Chronic Surgical Wound Infection. PLoS Genet 10(7): e32767. doi:10.1371/journal.pgen.1004518
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1004518
Souhrn
Soft tissue infections, such as those in burns, bed sores, and diabetic ulcers, are a significant healthcare and economic burden in the developed and developing world. The opportunistic pathogen P. aeruginosa can cause both acute and chronic infections, and the trajectory of these two types of infections is vastly different. We used high-throughput sequencing to profile P. aeruginosa genome-wide gene expression and mutant fitness during mouse model acute and non-diabetic chronic wound infections. Using these data, we show that wounds are nutrient-rich growth environments in which long-chain fatty acids are a primary source of carbon and energy. We also show that the ability to travel along spatio-chemical gradients by chemotaxis is critical for bacterial fitness and virulence in acute but not chronic infections. Our results demonstrate the utility of simultaneous mutant fitness and gene expression profiling to discover critical functions in complex growth environments.
Zdroje
1. BodeyGP, BolivarR, FainsteinV, JadejaL (1983) Infections caused by Pseudomonas aeruginosa. Rev Infect Dis 5: 279–313.
2. DaviesCE, HillKE, WilsonMJ, StephensP, HillCM, et al. (2004) Use of 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis for analysis of the microfloras of healing and nonhealing chronic venous leg ulcers. J Clin Microbiol 42: 3549–3557.
3. LazarusGS, CooperDM, KnightonDR, MargolisDJ, PecoraroRE, et al. (1994) Definitions and guidelines for assessment of wounds and evaluation of healing. Arch Dermatol 130: 489–493.
4. SamsonD, LefevreF, AronsonN (2004) Wound-healing technologies: low-level laser and vacuum-assisted closure. Evid Rep Technol Assess (Summ) 111: 1–6.
5. McManusAT, MasonAD, McManusWF, PruittBA (1985) Twenty-five year review of Pseudomonas aeruginosa bacteremia in a burn center. Eur J Clin Microbiol 4: 219–223.
6. PotokarTS, AliS, ChamaniaS, ProwseS, WhitakerIS (2008) A global overview of burns research highlights the need for forming networks with the developing world. Burns 34: 3–5.
7. KravitzM (1993) Immune consequences of burn injury. AACN Clin Issues Crit Care Nurs 4: 399–413.
8. LoryS, MerighiM, HyodoM (2009) Multiple activities of c-di-GMP in Pseudomonas aeruginosa. Nucleic Acids Symp Ser (Oxf) 53: 51–52.
9. LapougeK, SchubertM, AllainFH, HaasD (2008) Gac/Rsm signal transduction pathway of gamma-proteobacteria: from RNA recognition to regulation of social behaviour. Mol Microbiol 67: 241–253.
10. DaltonT, DowdSE, WolcottRD, SunY, WattersC, et al. (2011) An in vivo polymicrobial biofilm wound infection model to study interspecies interactions. PLoS ONE 6: e27317.
11. RumbaughKP, GriswoldJA, IglewskiBH, HamoodAN (1999) Contribution of quorum sensing to the virulence of Pseudomonas aeruginosa in burn wound infections. Infect Immun 67: 5854–5862.
12. CroucherNJ, ThomsonNR (2010) Studying bacterial transcriptomes using RNA-seq. Curr Opin Microbiol 13: 619–624.
13. WestermannAJ, GorskiSA, VogelJ (2012) Dual RNA-seq of pathogen and host. Nat Rev Microbiol 10: 618–630.
14. MandlikA, LivnyJ, RobinsWP, RitchieJM, MekalanosJJ, et al. (2011) RNA-Seq-based monitoring of infection-linked changes in Vibrio cholerae gene expression. Cell Host Microbe 10: 165–174.
15. CattoirV, NarasimhanG, SkurnikD, AschardH, RouxD, et al. (2013) Transcriptional Response of Mucoid Pseudomonas aeruginosa to Human Respiratory Mucus. mBio 3: e00410–12.
16. GoodmanAL, McNultyNP, ZhaoY, LeipD, MitraRD, et al. (2009) Identifying genetic determinants needed to establish a human gut symbiont in its habitat. Cell Host Microbe 6: 279–289.
17. van OpijnenT, BodiKL, CamilliA (2009) Tn-seq: high-throughput parallel sequencing for fitness and genetic interaction studies in microorganisms. Nat Methods 6: 767–772.
18. LangridgeGC, PhanM-D, TurnerDJ, PerkinsTT, PartsL, et al. (2009) Simultaneous assay of every Salmonella Typhi gene using one million transposon mutants. Genome Res 19: 2308–2316.
19. van OpijnenT, CamilliA (2012) A fine scale phenotype-genotype virulence map of a bacterial pathogen. Genome Res 22: 2541–2251.
20. GallagherLA, ShendureJ, ManoilC (2011) Genome-scale identification of resistance functions in Pseudomonas aeruginosa using Tn-seq. mBio 2: e00315–10.
21. SkurnikD, RouxD, AschardH, CattoirV, Yoder-HimesD, et al. (2013) A comprehensive analysis of in vitro and in vivo genetic fitness of Pseudomonas aeruginosa using high-throughput sequencing of transposon libraries. PLoS Pathog 9: e1003582.
22. HardingKG, MorrisHL, PatelGK (2002) Science, medicine and the future: healing chronic wounds. BMJ 324: 160–163.
23. WattersC, DeLeonK, TrivediU, GriswoldJA, LyteM, et al. (2013) Pseudomonas aeruginosa biofilms perturb wound resolution and antibiotic tolerance in diabetic mice. Med Microbiol Immunol 202: 131–141.
24. KingJD, KocincovaD, WestmanEL, LamJS (2009) Review: Lipopolysaccharide biosynthesis in Pseudomonas aeruginosa. Innate Immun 15: 261–312.
25. McMorranBJ, MerrimanME, RombelIT, LamontIL (1996) Characterisation of the pvdE gene which is required for pyoverdine synthesis in Pseudomonas aeruginosa. Gene 176: 55–59.
26. HauserAR (2009) The type III secretion system of Pseudomonas aeruginosa: infection by injection. Nat Rev Microbiol 7: 654–665.
27. DurandE, BernadacA, BallG, LazdunskiA, SturgisJN, et al. (2003) Type II protein secretion in Pseudomonas aeruginosa: the pseudopilus is a multifibrillar and adhesive structure. J Bacteriol 185: 2749–2758.
28. ColvinKM, IrieY, TartCS, UrbanoR, WhitneyJC, et al. (2012) The Pel and Psl polysaccharides provide Pseudomonas aeruginosa structural redundancy within the biofilm matrix. Environ Microbiol 14: 1913–1928.
29. AroraSK, NeelyAN, BlairB, LoryS, RamphalR (2005) Role of motility and flagellin glycosylation in the pathogenesis of Pseudomonas aeruginosa burn wound infections. Infect Immun 73: 4395–4398.
30. BurrowsLL (2012) Pseudomonas aeruginosa twitching motility: type IV pili in action. Annu Rev Microbiol 66: 493–520.
31. JorthP, TrivediU, RumbaughK, WhiteleyM (2013) Probing bacterial metabolism during infection using high-resolution transcriptomics. J Bacteriol 195: 4991–4998.
32. MacneilLT, WalhoutAJ (2011) Gene regulatory networks and the role of robustness and stochasticity in the control of gene expression. Genome Res 21: 645–657.
33. Diaz-PerezAL, Roman-DovalC, Diaz-PerezC, CervantesC, Sosa-AguirreCR, et al. (2007) Identification of the aceA gene encoding isocitrate lyase required for the growth of Pseudomonas aeruginosa on acetate, acyclic terpenes and leucine. FEMS Microbiol Lett 269: 309–316.
34. Alvarez-OrtegaC, HarwoodCS (2007) Responses of Pseudomonas aeruginosa to low oxygen indicate that growth in the cystic fibrosis lung is by aerobic respiration. Mol Microbiol 65: 153–165.
35. SerinoL, ReimmannC, ViscaP, BeyelerM, ChiesaVD, et al. (1997) Biosynthesis of pyochelin and dihydroaeruginoic acid requires the iron-regulated pchDCBA operon in Pseudomonas aeruginosa. J Bacteriol 179: 248–257.
36. ChengPN, LamTL, LamWM, TsuiSM, ChengAW, et al. (2007) Pegylated recombinant human arginase (rhArg-peg5,000mw) inhibits the in vitro and in vivo proliferation of human hepatocellular carcinoma through arginine depletion. Cancer Res 67: 309–317.
37. KangY, NguyenDT, SonMS, HoangTT (2008) The Pseudomonas aeruginosa PsrA responds to long-chain fatty acid signals to regulate the fadBA5 beta-oxidation operon. Microbiology 154: 1584–1598.
38. MillerRM, TomarasAP, BarkerAP, VoelkerDR, ChanED, et al. (2008) Pseudomonas aeruginosa twitching motility-mediated chemotaxis towards phospholipids and fatty acids: specificity and metabolic requirements. J Bacteriol 190: 4038–4049.
39. RomeroP, KarpP (2003) PseudoCyc, a pathway-genome database for Pseudomonas aeruginosa. J Mol Microbiol Biotechnol 5: 230–239.
40. WolfgangMC, KulasekaraBR, LiangX, BoydD, WuK, et al. (2003) Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 100: 8484–8489.
41. BrownSA, PalmerKL, WhiteleyM (2008) Revisiting the host as a growth medium. Nat Rev Microbiol 6: 657–666.
42. WadhamsGH, ArmitageJP (2004) Making sense of it all: bacterial chemotaxis. Nat Rev Mol Cell Biol 5: 1024–1037.
43. HongCS, ShitashiroM, KurodaA, IkedaT, TakiguchiN, et al. (2004) Chemotaxis proteins and transducers for aerotaxis in Pseudomonas aeruginosa. FEMS Microbiol Lett 231: 247–252.
44. Primrose SB, Twyman RM (2007) Principles of gene manipulation and genomics. Malden, MA: Blackwell Pub.
45. WestSA, GriffinAS, GardnerA, DiggleSP (2006) Social evolution theory for microorganisms. Nat Rev Microbiol 4: 597–607.
46. HuseHK, KwonT, ZlosnikJEA, SpeertDP, MarcotteEM, et al. (2010) Parallel evolution in Pseudomonas aeruginosa over 39,000 generations in vivo. mBio 1: e00199–10.
47. JacobsMA, AlwoodA, ThaipisuttikulI, SpencerD, HaugenE, et al. (2003) Comprehensive transposon mutant library of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 100: 14339–14344.
48. Ausubel FM (2002) Short protocols in molecular biology: a compendium of methods from Current protocols in molecular biology. New York: Wiley.
49. RietschA, Vallet-GelyI, DoveSL, MekalanosJJ (2005) ExsE, a secreted regulator of type III secretion genes in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 102: 8006–8011.
50. HoangTT, Karkhoff-SchweizerRR, KutchmaAJ, SchweizerHP (1998) A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 212: 77–86.
51. GibsonDG, YoungL, ChuangRY, VenterJC, HutchisonCA3rd, et al. (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6: 343–345.
52. GoodmanAL, WuM, GordonJI (2011) Identifying microbial fitness determinants by insertion sequencing using genome-wide transposon mutant libraries. Nat Protoc 6: 1969–1980.
53. AndersS, HuberW (2010) Differential expression analysis for sequence count data. Genome Biol 11: R106.
54. WinsorGL, LamDK, FlemingL, LoR, WhitesideMD, et al. (2011) Pseudomonas Genome Database: improved comparative analysis and population genomics capability for Pseudomonas genomes. Nucleic Acids Res 39: D596–600.
55. LangmeadB, SalzbergSL (2012) Fast gapped-read alignment with Bowtie 2. Nat Methods 9: 357–359.
56. ZomerA, BurghoutP, BootsmaHJ, HermansPW, van HijumSA (2012) ESSENTIALS: software for rapid analysis of high throughput transposon insertion sequencing data. PLoS One 7: e43012.
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2014 Číslo 7
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Wnt Signaling Interacts with Bmp and Edn1 to Regulate Dorsal-Ventral Patterning and Growth of the Craniofacial Skeleton
- Novel Approach Identifies SNPs in and with Evidence for Parent-of-Origin Effect on Body Mass Index
- Hypoxia Adaptations in the Grey Wolf () from Qinghai-Tibet Plateau
- DNA Topoisomerase 1α Promotes Transcriptional Silencing of Transposable Elements through DNA Methylation and Histone Lysine 9 Dimethylation in