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

Lung Transplant Rejection Clinical Trial

— NGS-ACRL  

Official title:

Next Generation Sequencing for the Diagnosis of Acute Rejection in Patients Undergoing Lung Transplantation. Pilot Study for Comparative Longitudinal Descriptive Trial.

Clinical Trial Details — Status: Active, not recruiting

Administrative data

• NCT number: NCT05260372
• Other study ID #: 1883/2018
• Status: Active, not recruiting
• Start date: October 4, 2018
• Est. completion date: December 2022

Study information

• Verified date: February 2022
• Source: Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial 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.

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.

Recruitment information / eligibility

• Status: Active, not recruiting
• Enrollment: 55
• Est. completion date: December 2022
• Est. primary completion date: November 2022
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years to 65 Years

Eligibility

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

Related Conditions & MeSH terms

• Lung Transplant Rejection

Intervention

Diagnostic Test:
• Quantification of donor-derived cell-free DNA
Quantification of ddcfDNA on serial plasma samples during follow-up after lung transplantation

Locations

Country	Name	City	State
Italy	Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico	Milan

Sponsors (2)

Lead Sponsor	Collaborator
Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico	University of Milan

Country where clinical trial is conducted

Italy, 

References & Publications (13)

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. — View Citation

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. Review. — View Citation

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. — View Citation

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. — View Citation

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. Review. — View Citation

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

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. — View Citation

Report CRT, Italy, Rete nazionale trapianti, 2017

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. — View Citation

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

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

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. — View Citation

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

* Note: There are 13 references in all — Click here to view all references

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Proportion ddcfDNA	Proportion of the ddcfDNA on the recipient's plasma cell free DNA	1 year
Secondary	ddcfDNA-clinical data correlation	Clinical data on trans-bronchial lung biopsies, pulmonary functional tests, culture of bronchoalveolar lavage fluid, donor-specific anti-human leukocyte antigen antibodies.	1 year