Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen

Antibioottivastustuskykyisten Klebsiella pneumoniae -bakteerikantojen aiheuttamat infektiot ovat globaalisti kasvava ongelma, sillä ne saattavat olla resistenttejä kaikille kliinisessä käytössä oleville antibiooteille. Infektioiden hoitoon voidaan vaihtoehtoisesti hyödyntää bakteereja tappavia viruk...

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Main Author: Thind, Navjot
Other Authors: Matemaattis-luonnontieteellinen tiedekunta, Faculty of Sciences, Bio- ja ympäristötieteiden laitos, Department of Biological and Environmental Science, Jyväskylän yliopisto, University of Jyväskylä
Format: Master's thesis
Language:fin
Published: 2021
Subjects:
Online Access: https://jyx.jyu.fi/handle/123456789/79417
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author Thind, Navjot
author2 Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_facet Thind, Navjot Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä Thind, Navjot Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_sort Thind, Navjot
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description Antibioottivastustuskykyisten Klebsiella pneumoniae -bakteerikantojen aiheuttamat infektiot ovat globaalisti kasvava ongelma, sillä ne saattavat olla resistenttejä kaikille kliinisessä käytössä oleville antibiooteille. Infektioiden hoitoon voidaan vaihtoehtoisesti hyödyntää bakteereja tappavia viruksia, bakteriofageja, joita käytetään infektioiden hoidoissa sellaisenaan tai niiden tuottamien lyyttisten entsyymien kautta. Tämän tutkimuksen tavoitteena on ymmärtää kahden K. pneumoniae -kannan (EKP3 ja EKP8) ja Nenäinniemen jätevedenpuhdistamosta eristettyjen faagien välistä vuorovaikutusta, sekä löytää uusia näkökulmia faagiterapian kehittämiseksi. EKP3 ja EKP8 villityyppikantojen lisäksi molemmista kannoista tutkittavana oli kaksi mutanttia isolaattia, jotka olivat kehittyneet resistenteiksi kahdeksalle ennestään eristetyille faageille. Kannat erosivat toisistaan geneettisesti siten, että ainoastaan toisella kannoista (EKP3:lla) on CRISPR-lokuksia. Tutkimuksen aikana suoritetut faagialtistukset paljastivat EKP3-kannan reagoivan faageihin geneettisellä tasolla, toisin kuin EKP8, joka reagoi voimakkaasti muokkaamalla geenisäätelyä. Uusien, faagiterapian kannalta, potentiaalisten faagien eristäminen jätevedestä onnistui hyvin. Erityisen kiinnostavaa on EKP8P6- faagin kyky infektoida EKP3-kantaa vain silloin, kun se ensin oli kehittänyt resistenssin EKP8P3-faagia vastaan. Vastaavia resistenttejä muotoja infektoivia faageja altistuksiin lisäämällä voidaan mahdollisesti parantaa faagiterapian onnistumistodennäköisyyttä ja hoidon tehokkuutta tulevaisuudessa. Faagialtistuskokeet eivät laboratorio-oloissa aktivoineet isännän CRISPR-järjestelmää, mitä voidaan pitää myös lupaavana faagiterapian kannalta. Antibiotics have been used for decades to treat bacterial infections, such as ones caused by Klebsiella pneumoniae, a well-known multi-drug resistant pathogen. The efficacy of antibiotics in the treatment of bacterial infections has, however, been weakened by the growing prevalence of antibiotic resistance genes in the bacterial population. Bacteriophages, or phages for short, are viruses which specifically infect bacteria and therefore could act as an alternative for antibiotic treatments in the form of phage therapy. The purpose of this study was to examine the responses of two K. pneumoniae strains (EKP3 and EKP8) to isolated phages to further improve our knowledge on phage therapy. Both strains had two mutant isolates that were previously treated with eight phages and had developed resistances against them. These strains differed from each other genetically by CRISPR-loci that only EKP3 possessed, though it wasn´t activated during any of the experiments. Results showed how EKP3 altered its genome to adapt to environmental changes and phage pressure, unlike EKP8, which altered its gene expression instead. We were able to isolate promising novel phages from sewage water, especially EKP8P6, which was able to infect EKP3 only when the bacterium had developed resistance against another phage isolated from sewage previously. This study shows new insights into how we could insert phages like EKP8P6 that cannot infect wild-type bacteria but rather mutated ones, into phage cocktails to improve phage therapy treatments against pathogenic bacteria like K. pneumoniae.
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spellingShingle Thind, Navjot Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen CRISPR infektiokoe Solu- ja molekyylibiologia Cell and molecular biology 4013 fagiterapia mutaatiot bakteerit resistenssi mikrobiologia bakteriofagit infektiot antibiootit DNA taudinaiheuttajat Klebsiella-bakteerit
title Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
title_full Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
title_fullStr Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
title_full_unstemmed Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
title_short Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
title_sort klebsiella pneumoniae bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
title_txtP Klebsiella pneumoniae -bakteerilajia infektoivien faagien eristys ja faagiresistenssimekanismien kartoittaminen
topic CRISPR infektiokoe Solu- ja molekyylibiologia Cell and molecular biology 4013 fagiterapia mutaatiot bakteerit resistenssi mikrobiologia bakteriofagit infektiot antibiootit DNA taudinaiheuttajat Klebsiella-bakteerit
topic_facet 4013 CRISPR Cell and molecular biology DNA Klebsiella-bakteerit Solu- ja molekyylibiologia antibiootit bakteerit bakteriofagit fagiterapia infektiokoe infektiot mikrobiologia mutaatiot resistenssi taudinaiheuttajat
url https://jyx.jyu.fi/handle/123456789/79417 http://www.urn.fi/URN:NBN:fi:jyu-202201191185
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