Effect of Adenovirus E1A Oncogene on DNA Replication Dynamics

NCT03325517 | Unknown

• 1 other identifier: 17200086.
• Study Type: observational
• Target: 50
• Locations: 0 countries
• Sites: N/A

Brief Summary

BACKGROUND: The 243 amino acid E1A encoded by the left end of the human adenovirus (Ad) type 2 or 5 genome has been studied in various contexts including as a model cooperating oncoprotein , an apoptosis inducing protein, and as a therapeutic oncolytic protein. All of these properties are associated with its capacity to rapidly induce S phase in a variety of cells. E1A orchestrates most of these effects by interacting with an array of chromatin remodeling complexes, including the Rb family proteins, and the HAT proteins p300 and CBP. The Myst family protein HBO1 (Myst2, KAT7) is a histone acetyl transferase that plays a major role in replication initiation and also contributes to DNA re-replication. HBO1 directly interacts with Cdt1 and functions as a coactivator of Cdt1 in replication initiation. It also associates with replication origin and stimulates origin activation by acetylating H4 K5, K8, and K12. Overexpression of HBO1 induces DNA re-replication. SPECIFIC AIM OF THE STUDY: The Specific Aim of this study is to determine whether or not the stimulation of HBO1 activity by E1A plays a role in deregulated DNA replication, and if it does, to determine the mechanism.

1. 1.Using standard assays the investigators will determine whether E1A binds to HBO1 to induce its HAT activity
2. 2.The investigators will determine whether E1A stimulation of HAT activity of HBO1 contributes to DNA re-replication.

Conditions

Adenovirus

Outcome Measures

Primary Outcomes (1)

• DNA re-replication indicated by the number of cells containing >4N DNA content using flow cytometer

  To determine whether E1A uses HBO1 in inducing re-replication by stimulating the HBO1 HAT activity, the investigators will infect S phase enriched cells (isolated by double thymidine block) with Ad viruses expressing the WT or mutant E1A proteins along with Ad vectors expressing HBO1 variants (e.g. HAT inactive), allow the infection to proceed as needed then quantify the population of cells containing \>4N DNA content using flow cytometer. If E1A simulated HBO1 HAT activity is important for re-replication HAT defective HBO1 mutants will not induce re-replication even in the presence of E1A. E1A mutants that cannot bind to HBO1 will also show a similar phenotype.

  1 year

Eligibility Criteria

• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

All patients who are positive for adenovirus (symptomatic or not)

You may qualify if:

• \- All patients who are positive for adenovirus

You may not qualify if:

• \- immune comprised patients

Sponsors & Collaborators

• Heba Momen kamel: lead

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Biospecimen

Retention: SAMPLES WITH DNA

cell lines

MeSH Terms

Conditions

Adenoviridae Infections

Condition Hierarchy (Ancestors)

DNA Virus Infections; Virus Diseases; Infections

Study Design

• Study Type: observational
• Observational Model: CASE ONLY
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: SPONSOR INVESTIGATOR
• PI Title: Assistant Lecturer

Study Record Dates

• First Submitted: October 24, 2017
• First Posted: October 30, 2017
• Study Start: November 1, 2017
• Primary Completion: September 1, 2018
• Study Completion: September 1, 2019
• Last Updated: October 30, 2017

Record last verified: 2017-10