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Status unknownNCT03342547Updated Sep 20, 2018

Identification of Host Factors of Norovirus Infections in Mini-Gut Model

An interventional study of Duodenal biopsy and Saliva in Gastrointestinal Infection, sponsored by Chinese University of Hong Kong. Status unknown at 1 site in China. Open to participants aged 18 Years and older, including healthy volunteers. Per ClinicalTrials.gov, last updated 2018-09-20.

Sponsored by Chinese University of Hong Kong · Not applicable, Interventional, and Basic science

The sponsor has not verified this record recently (last verified Sep 2018), so the status shown — last known as Recruiting — may be out of date.
Phase
Not applicable
Study type
Interventional
Enrollment
20
Allocation
Not applicable
Ages
18 Years and older
Sex
All
01

Study summary

The primary objective in this study is to establish a list of host cellular proteins that mediate norovirus infection.

Norovirus is one of the most common pathogens attributed to diarrheal diseases from unsafe food. It is also the primary cause of mortality among young children and adults in foodborne infections. Norovirus is not just a foodborne burden. In a recent meta-analysis, norovirus accounts for nearly one-fifth of all causes of (including person-to-person transmission) acute gastroenteritis in both sporadic and outbreak settings and affects all age groups. Undoubtedly, norovirus is of paramount public health concern in both developed and developing countries. Research efforts to better understand norovirus pathobiology will be necessary for targeted intervention.

From Middle East respiratory syndrome coronavirus to Zika virus, efforts to identify host factors important for mediating virus infection has always been a research priority. Such information will shed light on potential therapeutic targets in antiviral intervention. Norovirus virus-host interaction studies have been hampered by the lack of a robust cell culture model in the past 20 years. In 2016, norovirus has finally been successfully cultivated in a stem cell-derived three-dimensional human gut-like structure called enteroid or mini-gut.

In this study, intestinal stem cells will be isolated from duodenal biopsies collected from participants, followed by differentiation into mini-guts. Genome-wide genetic screening for host essential and restrictive factors will be performed on infected mini-guts by knockout CRISPR and gain-of-function CRISPR SAM, respectively. Shortlisted candidates will undergo preliminary functional validation in cell lines. These data will provide insights into potential therapeutic targets against norovirus infection.

Read the detailed description

Study design and overview - This is a laboratory-based study. The investigators plan to establish a list of host cellular proteins regulating norovirus infection. Methods will include establishment of patient-specific stem cell-derived human intestinal enteroids as an infection model and genome-wide CRISPR and CRISPR SAM genetic screens upon infections of two norovirus genotypes.

***Establishment of human intestinal enteroid model***

  1. Duodenal biopsies and saliva collection - After obtaining IRB-approved informed consent, two duodenal biopsy samples will be obtained from each participant undergoing upper gastrointestinal endoscopy without contraindications for biopsy sampling, such as participants on anti-coagulation or with other causes of bleeding diathesis. Biopsy samples will be immersed in ice-cold 1xPBS and transported immediately to the laboratory within 30 minutes for further processing. In addition, saliva (1-2 mL) will be collected from each participant for secretor status testing by ELISA.
  2. Isolation of crypt stem cells and differentiation - Upon arrival, biopsy samples will first be chopped into smaller pieces. Intestinal crypt cells will be isolated by repeated washing and incubation with 1x Complete Chelating solution (1xCCS) and EDTA buffer. Supernatant containing crypt cells will then be grown in Matrigel containing complete medium with growth factors (CMGF+) (Wnt 3 and Rspo-1 conditioned media, Noggin, A83, B27, EGF, N2, n-acetylcysteine, gastrin, nicotinamide, and SB202190). Budding crypts will then be incubated in differentiation medium (CMGF+, excluding Wnt 3 conditioned medium, SB202190 and nicotinamide as well as having reduced amount of Rspo-1 conditioned medium and Noggin). Formation of three-dimensional gut-like enteroids will be monitored daily.
02

Conditions studied

  • Gastrointestinal Infection

Keywords

  • Norovirus
03

Who can participate

Ages eligible
18 Years and older
Sexes eligible
All
Accepts healthy volunteers
Yes

Inclusion criteria

  • Adults aged 18 years or above
  • Prospective outpatients undergoing gastrointestinal endoscopy for symptoms of dyspepsia in the Endoscopy Unit of the Prince of Wales Hospital, Shatin, Hong Kong, China
  • Able and willing to provide informed written consent

Exclusion criteria

Exclusion Criteria:

  • Use of anti-coagulants and/or aspirin that may have increased risk of bleeding
  • History of bleeding diathesis
  • Contraindications for biopsy sampling
04

Study design

Phase
Not applicable
Primary purpose
Basic science
Allocation
Not applicable
Intervention model
Single group
Masking
None (open label)
Enrollment
20 participants (estimated)

Study arms

  • Experimental
    Intestinal stem cell-derived enteroids

    Duodenal biopsy samples and saliva will be collected from participants and then processed in laboratory to generate intestinal stem cell-derived enteroids.

    Procedure: Duodenal biopsy · Procedure: Saliva

Interventions

  • ProcedureDuodenal biopsy

    Two duodenal biopsy samples will be collected from each participant

  • ProcedureSaliva

    Saliva (1-2 mL) will be collected from each participant for secretor status testing

05

What researchers measure

Primary outcomes

  1. Establishment of human intestinal stem cell-derived enteroids

    Viability of enteroids as determined by microscopy

    Time frame: An average of three months

06

Study locations

1 of 1 sites recruiting
  • Endoscopy Centre, Prince of Wales Hospital
    Hong Kong, China
    • Chi-Wai Chan, PhD · Contact · martin.chan@cuhk.edu.hk · +852 35051523
    • Chi-Wai Chan, PhD · Principal investigator
    • Kay-Sheung Chan, MD · Sub investigator
    • Hei Wong, MBChB; DPhil · Sub investigator
    Recruiting
07

References and documents

Individual participant data

Plan to share: Undecided

No publications or documents are linked to this record.

08

Registry details

Key details

Study ID
NCT03342547
Lead sponsor
Chinese University of Hong Kong
Responsible party
CHAN CHI WAI (Assistant Professor, Chinese University of Hong Kong) — Principal investigator
First posted
Nov 17, 2017
Start date
Apr 18, 2018
Primary completion
Dec 31, 2020 (estimated)
Completion
Dec 31, 2020 (estimated)
Last update
Sep 20, 2018

Study contacts

Chi-Wai Chan, PhD
Contact
martin.chan@cuhk.edu.hk
+852 35051523
Chi-Wai Chan, PhD
principal investigator · Chinese University of Hong Kong

Oversight

Data monitoring committee
No
FDA-regulated drug
No
FDA-regulated device
No
View the source record on ClinicalTrials.gov ↗

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