Next Generation Sequencing to Detect Acute Rejection in Lung Transplant Patients.

NCT ID: NCT05260372

Last Updated: 2022-03-02

Basic Information

• Recruitment Status: UNKNOWN
• Total Enrollment: 55 participants
• Study Classification: OBSERVATIONAL
• Study Start Date: 2018-10-04
• Study Completion Date: 2022-12-31

Brief Summary

Lung transplantation is a consolidated treatment in selected patients with end-stage respiratory diseases; however, acute rejection remains an important cause of lung allograft loss and a risk factor for chronic allograft dysfunction. Histopathological examination of lung tissue is the gold standard for the diagnosis of acute rejection, therefore recipients undergo surveillance transbronchial biopsy and bronchoalveolar lavage after transplantation. However, the obtained tissue is sometimes inadequate for histopathology, and the endoscopic procedure can lead to complications (bleeding, pneumothorax). The quantification of donor-derived cell-free DNA (ddcfDNA) in the recipient plasma has shown to be increased in case of acute rejection, and could represent an early and non-invasive diagnostic marker to detect acute rejection. We planned to enroll all patients aged 18 to 65 years old enlisted for lung transplantation at our centre. Patients undergoing retransplantation and patients with a history of prior solid organ transplantation were excluded. The quantification of donor-derived cell-free DNA was performed 15 days and 3, 6, and 12 months after transplantation, concurrently with the routine surveillance bronchoscopies as per our protocol; the same analysis was also conducted in case of suspected clinical rejection.

Detailed Description

The purpose of the study is the identification of a non-invasive post-transplantation acute lung rejection monitoring method, by identification of donor derived cell free DNA (ddcf-DNA) in the recipient's plasma. At the time of transplantation, the ddcf-DNA derived from lymphocytes present in the peripheral blood of the donor and recipient will be genotyped by analysis of about 300 single nucleotide polymorphisms (SNPs) with a high probability of being differently homozygous between donor and recipient, according to the frequencies in the population reported on genomic databases. During the post-transplant follow-up, the presence of ddcf-DNA will be monitored by identification of the donor's SNP in the free DNA extracted from the recipient's plasma using the next-generation sequencing (NGS) method. Levels of ddcfDNA in the recipient will be correlated with clinical and histological data obtained from routine 3, 6 and 12 month post-transplant surveillance lung biopsies. All patients enrolled in the study will sign a study-specific written informed consent.

-Timeline: T-1 (before lung transplantation): DNA genotyping of the recipient; T0 (lung transplantation): genotyping of donor DNA; T1, T2, T3, T4 (15 days, 3, 6, 12 months after lung transplantation): ddcfDNA research on recipient serum; Tr: in any case of clinical suspicion of acute cellular rejection. At T2, T3, T4 and Tr, surveillance trans-bronchial lung biopsies (TBBs) are also performed, as per our centre's standard post-transplant lung follow-up protocol.

-Sample size and statistical analysis: Proportion of ddcfDNA/recipient DNA: stable patients (pA): 0.01, patients with graft damage (pB): 0.30, proportion nA/nB: 3, Power (1-beta): 0.80, Alpha: 5%, Margin of non inferiority δ: 0.05. Population (A+B) : 48; for possible exit from the study we add 7 cases for a total of 55 patients.

Conditions

Lung Transplant Rejection

Study Design

• Observational Model Type: COHORT
• Study Time Perspective: PROSPECTIVE

Study Groups

Lung transplantation recipients

Recipients aged 18 to 65 years old undergoing lung transplantation at Fondazione IRCCS Ospedale Policlinico Milano undergoing quantification of donor-derived cell-free DNA on serial plasma samples

Quantification of donor-derived cell-free DNA

Intervention Type: DIAGNOSTIC_TEST

Quantification of ddcfDNA on serial plasma samples during follow-up after lung transplantation

Interventions

Quantification of donor-derived cell-free DNA

Quantification of ddcfDNA on serial plasma samples during follow-up after lung transplantation

Intervention Type: DIAGNOSTIC_TEST

Eligibility Criteria

Inclusion Criteria

• Patients undergoing lung transplantation at our centre
• Patients aged 18 to 65 years old
• Written informed consent to partecipate in the study

Exclusion Criteria

• Patients undergoing retransplantation
• Patients with a history of prior solid organ transplantation

• Minimum Eligible Age: 18 Years
• Maximum Eligible Age: 65 Years
• Eligible Sex: ALL
• Accepts Healthy Volunteers: No

Sponsors

University of Milan

OTHER

Sponsor Role: collaborator

Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico

OTHER

Sponsor Role: lead

Responsible Party

Mario Nosotti

Full professor, Chief of Thoracic Surgery and Lung transplantation Unit

Responsibility Role: PRINCIPAL_INVESTIGATOR

Principal Investigators

Mario Nosotti, MD, Professor

Role: PRINCIPAL_INVESTIGATOR

Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico

Locations

Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico

Milan, , Italy

Countries

Italy

References

Report CRT, Italy, Rete nazionale trapianti, 2017

Reference Type: BACKGROUND

Chambers DC, Yusen RD, Cherikh WS, Goldfarb SB, Kucheryavaya AY, Khusch K, Levvey BJ, Lund LH, Meiser B, Rossano JW, Stehlik J; International Society for Heart and Lung Transplantation. The Registry of the International Society for Heart and Lung Transplantation: Thirty-fourth Adult Lung And Heart-Lung Transplantation Report-2017; Focus Theme: Allograft ischemic time. J Heart Lung Transplant. 2017 Oct;36(10):1047-1059. doi: 10.1016/j.healun.2017.07.016. Epub 2017 Jul 19. No abstract available.

Reference Type: BACKGROUND

PMID: 28784324 (View on PubMed)

Lande JD, Patil J, Li N, Berryman TR, King RA, Hertz MI. Novel insights into lung transplant rejection by microarray analysis. Proc Am Thorac Soc. 2007 Jan;4(1):44-51. doi: 10.1513/pats.200605-110JG.

Reference Type: BACKGROUND

PMID: 17202291 (View on PubMed)

Tosi D, Carrinola R, Morlacchi LC, Tarsia P, Rossetti V, Mendogni P, Rosso L, Righi I, Damarco F, Nosotti M. Surveillance Transbronchial Biopsy Program to Evaluate Acute Rejection After Lung Transplantation: A Single Institution Experience. Transplant Proc. 2019 Jan-Feb;51(1):198-201. doi: 10.1016/j.transproceed.2018.04.073. Epub 2018 Jun 28.

Reference Type: BACKGROUND

PMID: 30655138 (View on PubMed)

Rademacher J, Suhling H, Greer M, Haverich A, Welte T, Warnecke G, Gottlieb J. Safety and efficacy of outpatient bronchoscopy in lung transplant recipients - a single centre analysis of 3,197 procedures. Transplant Res. 2014 May 27;3:11. doi: 10.1186/2047-1440-3-11. eCollection 2014.

Reference Type: BACKGROUND

PMID: 24917927 (View on PubMed)

De Vlaminck I, Martin L, Kertesz M, Patel K, Kowarsky M, Strehl C, Cohen G, Luikart H, Neff NF, Okamoto J, Nicolls MR, Cornfield D, Weill D, Valantine H, Khush KK, Quake SR. Noninvasive monitoring of infection and rejection after lung transplantation. Proc Natl Acad Sci U S A. 2015 Oct 27;112(43):13336-41. doi: 10.1073/pnas.1517494112. Epub 2015 Oct 12.

Reference Type: BACKGROUND

PMID: 26460048 (View on PubMed)

Agbor-Enoh S, Jackson AM, Tunc I, Berry GJ, Cochrane A, Grimm D, Davis A, Shah P, Brown AW, Wang Y, Timofte I, Shah P, Gorham S, Wylie J, Goodwin N, Jang MK, Marishta A, Bhatti K, Fideli U, Yang Y, Luikart H, Cao Z, Pirooznia M, Zhu J, Marboe C, Iacono A, Nathan SD, Orens J, Valantine HA, Khush K. Late manifestation of alloantibody-associated injury and clinical pulmonary antibody-mediated rejection: Evidence from cell-free DNA analysis. J Heart Lung Transplant. 2018 Jul;37(7):925-932. doi: 10.1016/j.healun.2018.01.1305. Epub 2018 Jan 31.

Reference Type: BACKGROUND

PMID: 29500138 (View on PubMed)

De Vlaminck I, Valantine HA, Snyder TM, Strehl C, Cohen G, Luikart H, Neff NF, Okamoto J, Bernstein D, Weisshaar D, Quake SR, Khush KK. Circulating cell-free DNA enables noninvasive diagnosis of heart transplant rejection. Sci Transl Med. 2014 Jun 18;6(241):241ra77. doi: 10.1126/scitranslmed.3007803.

Reference Type: BACKGROUND

PMID: 24944192 (View on PubMed)

Rizopoulos, D. Joint Models for Longitudinal and Time-to-Event Data, with Applications in R. Boca Raton: Chapman & Hall/CRC. 2012.

Reference Type: BACKGROUND

Wu, L. Mixed Effects Models for Complex Data. Boca Raton: Chapman & Hall/CRC, Chapter 8. 2009

Reference Type: BACKGROUND

Rizopoulos, D. The R package JMbayes for fitting joint models for longitudinal and timeto-event data using MCMC. Journal of Statistical Software 72(7), 1-45. doi:10.18637/jss.v072.i07. 2016

Reference Type: BACKGROUND

Rizopoulos D. Dynamic predictions and prospective accuracy in joint models for longitudinal and time-to-event data. Biometrics. 2011 Sep;67(3):819-29. doi: 10.1111/j.1541-0420.2010.01546.x. Epub 2011 Feb 9.

Reference Type: BACKGROUND

PMID: 21306352 (View on PubMed)

R Foundation for Statistical Computing, Vienna, Austria(2015)

Reference Type: BACKGROUND

Other Identifiers

1883/2018

Identifier Type: -

Identifier Source: org_study_id