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Implementation of microbiota analysis in clinical trials for cystic fibrosis lung infection: experience from the OligoG phase 2b clinical trials

Implementation of microbiota analysis in clinical trials for cystic fibrosis lung infection: experience from the OligoG phase 2b clinical trials
Implementation of microbiota analysis in clinical trials for cystic fibrosis lung infection: experience from the OligoG phase 2b clinical trials

Culture-independent microbiota analysis is widely used in research and being increasingly used in translational studies. However, methods for standardisation and application of these analyses in clinical trials are limited. Here we report the microbiota analysis that accompanied two phase 2b clinical trials of the novel, non-antibiotic therapy OligoG CF-5/20 for cystic fibrosis (CF) lung infection. Standardised protocols (DNA extraction, PCR, qPCR and 16S rRNA gene sequencing analysis) were developed for application to the Pseudomonas aeruginosa (NCT02157922) and Burkholderia cepacia complex (NCT02453789) clinical trials involving 45 and 13 adult trial participants, respectively. Microbiota analysis identified that paired sputum samples from an individual participant, taken within 2 h of each other, had reproducible bacterial diversity profiles. Although culture microbiology had identified patients as either colonised by P. aeruginosa or B. cepacia complex species at recruitment, microbiota analysis revealed patient lung infection communities were not always dominated by these key CF pathogens. Microbiota profiles were patient-specific and remained stable over the course of both clinical trials (6 sampling points over the course of 140 days). Within the Burkholderia trial, participants were infected with B. cenocepacia (n = 4), B. multivorans (n = 6), or an undetermined species (n = 3). Colonisation with either B. cenocepacia or B. multivorans influenced the overall bacterial community structure in sputum. Overall, we have shown that sputum microbiota in adults with CF is stable over a 2 h time-frame, suggesting collection of a single sample on a collection day is sufficient to capture the microbiota diversity. Despite the uniform pathogen culture-positivity status at recruitment, trial participants were highly heterogeneous in their lung microbiota. Understanding the microbiota profiles of individuals with CF ahead of future clinical trials would be beneficial in the context of patient stratification and trial design.

Clinical trials, Cystic fibrosis microbiology, Microbiota analysis, PCR, qPCR and 16S rRNA gene sequencing
0167-7012
106133
Weiser, Rebecca
0091ccfb-41be-4c40-94f7-f1382733d691
Rye, Philip D.
823bccb7-4c2a-4324-bcff-d0ffc950365d
Mahenthiralingam, Eshwar
ace82a27-cc14-46de-8352-b0b98c477d93
Carroll, Mary
b836d262-6b07-4006-9c81-26653a26588b
Conway, Joy H
bbe9a2e4-fb85-4d4a-a38c-0c1832c32d06
OligoG phase 2b clinical trials group
Weiser, Rebecca
0091ccfb-41be-4c40-94f7-f1382733d691
Rye, Philip D.
823bccb7-4c2a-4324-bcff-d0ffc950365d
Mahenthiralingam, Eshwar
ace82a27-cc14-46de-8352-b0b98c477d93
Carroll, Mary
b836d262-6b07-4006-9c81-26653a26588b
Conway, Joy H
bbe9a2e4-fb85-4d4a-a38c-0c1832c32d06

Weiser, Rebecca, Rye, Philip D. and Mahenthiralingam, Eshwar , OligoG phase 2b clinical trials group (2021) Implementation of microbiota analysis in clinical trials for cystic fibrosis lung infection: experience from the OligoG phase 2b clinical trials. Journal of Microbiological Methods, 181, 106133, [106133]. (doi:10.1016/j.mimet.2021.106133).

Record type: Article

Abstract

Culture-independent microbiota analysis is widely used in research and being increasingly used in translational studies. However, methods for standardisation and application of these analyses in clinical trials are limited. Here we report the microbiota analysis that accompanied two phase 2b clinical trials of the novel, non-antibiotic therapy OligoG CF-5/20 for cystic fibrosis (CF) lung infection. Standardised protocols (DNA extraction, PCR, qPCR and 16S rRNA gene sequencing analysis) were developed for application to the Pseudomonas aeruginosa (NCT02157922) and Burkholderia cepacia complex (NCT02453789) clinical trials involving 45 and 13 adult trial participants, respectively. Microbiota analysis identified that paired sputum samples from an individual participant, taken within 2 h of each other, had reproducible bacterial diversity profiles. Although culture microbiology had identified patients as either colonised by P. aeruginosa or B. cepacia complex species at recruitment, microbiota analysis revealed patient lung infection communities were not always dominated by these key CF pathogens. Microbiota profiles were patient-specific and remained stable over the course of both clinical trials (6 sampling points over the course of 140 days). Within the Burkholderia trial, participants were infected with B. cenocepacia (n = 4), B. multivorans (n = 6), or an undetermined species (n = 3). Colonisation with either B. cenocepacia or B. multivorans influenced the overall bacterial community structure in sputum. Overall, we have shown that sputum microbiota in adults with CF is stable over a 2 h time-frame, suggesting collection of a single sample on a collection day is sufficient to capture the microbiota diversity. Despite the uniform pathogen culture-positivity status at recruitment, trial participants were highly heterogeneous in their lung microbiota. Understanding the microbiota profiles of individuals with CF ahead of future clinical trials would be beneficial in the context of patient stratification and trial design.

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More information

Accepted/In Press date: 4 January 2021
e-pub ahead of print date: 7 January 2021
Published date: February 2021
Additional Information: Funding Information: The following investigators and their teams (in alphabetical order) were involved in the clinical trials. Mary Carroll, General Hospital, Southampton, UK; Jane Davies, Royal Brompton and Harefield NHS Foundation Trust, London, UK (the study was supported by the NIHR Biomedical Research Unit and Clinical Research Facility); Carsten Schwarz and Nico Derichs, Christiane Herzog Zentrum, Charité Berlin, Germany; Damian Downey, Centre for Experimental Medicine, Queen's University, Belfast, UK; Olaf Eickmeier, University Hospital, Frankfurt a.M., Germany; Pal Leyell Finstad, University Hospital, Oslo, Norway; Rainald Fischer, Pneumologische Praxis Pasing, Munich, Germany; Marie Øbro Fossbøl, Rigshospitalet, Copenhagen, Denmark; Marita Gilljam, Sahlgrenska Universitetssjukhuset, Göteborg, Sweden; Charles Haworth, Papworth Hospital, Cambridge, UK; Lena Hjelte, Karolinska Universitetssjukhuset, Stockholm, Sweden; Alan Knox, City Hospital, Nottingham, UK; Silke van Koningsbruggen-Rietschel, University Hospital, Cologne, Germany; Gordon MacGregor, Queen Elizabeth University Hospital, Glasgow, UK; Jann Mortensen, Rigshospitalet, Copenhagen, Denmark; Susanne Nährig, LMU, Munich, Germany; Tacjana Pressler, Rigshospitalet, Copenhagen, Denmark; Joachim Riethmüller, University Hospital, Tübingen, Germany; Felix C. Ringshausen, MHH, Hannover, Germany; Martin Walshaw, Heart and Chest Hospital, Liverpool, UK; qualified person responsible for pharmacovigilance: Hugo Flaten, AlgiPharma AS, Sandvika, Norway; statistician: Nils Meland, Smerud Medical Research International AS, Oslo, Norway. The MCC substudy was performed at three sites: Laura Gow, Bio-Images Research Ltd., Glasgow, UK; Joy Conway, University of Southampton, Southampton, UK; Jann Mortensen, Marie Øbro Fosbøl, Rigshospitalet, Copenhagen, Denmark. Data was evaluated and reviewed by Scott H. Donaldson and William Bennett, UNC Chapel Hill, NC, USA. Funding Information: Funding for this study was received from the Cystic Fibrosis Foundation, Bethesda, MD, USA, with additional funding provided by the study sponsor AlgiPharma AS, Sandvika, Norway. The study sponsor representative (PDR) participated in the study design, data collection, data analysis, data interpretation and writing of the study report. The authors had full access to all data and had final responsibility for publication. The final decision on content was exclusively retained by the contributing authors. EM and RW acknowledge current funding by the Cystic Fibrosis Foundation (award MAHENT20G0 ). Publisher Copyright: © 2021 Elsevier B.V. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
Keywords: Clinical trials, Cystic fibrosis microbiology, Microbiota analysis, PCR, qPCR and 16S rRNA gene sequencing

Identifiers

Local EPrints ID: 450416
URI: http://eprints.soton.ac.uk/id/eprint/450416
ISSN: 0167-7012
PURE UUID: e3ae1d13-712f-4ed1-9ee2-8f4913829261
ORCID for Joy H Conway: ORCID iD orcid.org/0000-0001-6464-1526

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Date deposited: 27 Jul 2021 17:25
Last modified: 17 Mar 2024 12:47

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Contributors

Author: Rebecca Weiser
Author: Philip D. Rye
Author: Eshwar Mahenthiralingam
Author: Mary Carroll
Author: Joy H Conway ORCID iD
Corporate Author: OligoG phase 2b clinical trials group

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