# Project 1: KSHV reprograms replication and metabolic activities in hypoxia

> **NIH NIH P01** · UNIVERSITY OF PENNSYLVANIA · 2024 · $468,525

## Abstract

Project #1
Abstract
KSHV-associated cancers occur in hypoxic microenvironments. The effects of hypoxia on viral and host DNA
replication in the context of KSHV infection are poorly understood. We have previously investigated the replication
of KSHV in normoxia cell culture systems. We now propose an in-depth analysis of KSHV and infected host cell
replication in the hypoxic environment. KSHV, like many members of the herpesvirus family undergoes both a
latent and a lytic replication cycle. In cell culture systems, latency is maintained through synchronization of the
viral genome replication with that of the host cell replication, which is cell cycle dependent. Therefore, most of the
latent replication activities utilize the host machinery once per cell cycle. These latently infected cells are typically
induced to lytic replication through treatment with chemical inducers and requires the expression of a number of
viral encoded genes. However, little is known regarding replication of the virus genome in hypoxia, and the viral
antigens needed to regulate the ongoing metabolic changes required for persistence of the viral genome through
replication in infected cells that allow for propagation into new daughter cells. We will approach these questions
through a number of aims geared towards understanding the changes in the cellular and viral replication proteins
that occur in hypoxia compared to that seen in normoxia. We will examine the changes in cellular replication
proteins at different phases of the cell cycle, and their post-translational modification. Their levels in hypoxia will
be analyzed to determine the changes in the essential components of the replication machinery. In addition, we
will look at their transcription to determine if these changes are due to changes in the overall transcription of the
genes or if the changes are due to transcription modulation by HIF1α in hypoxia. The changes in metabolic
enzymes that are induced in KSHV infected cells compared to KSHV negative cells in hypoxia will also be analyzed
to determine if the changes are transcriptionally reprogrammed. We will determine the metabolic genes that are
dysregulated on KSHV infection in hypoxia and if the replication genes are regulated by HIF1α, the master
transcription regulator in hypoxia. We will determine the epigenetic changes that occur during hypoxia that will
determine the expression of the replication genes. We will also perform these assays in 3D culture systems to
mimic the associated pathologies. We will also generate shRNA knockdown or CRISPR knockout of selected
replication genes, and Tet-regulated genes to determine the contribution of these proteins in KSHV replication in
the hypoxic microenvironment.
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## Key facts

- **NIH application ID:** 10914914
- **Project number:** 5P01CA281867-02
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** ERLE S. ROBERTSON
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $468,525
- **Award type:** 5
- **Project period:** 2023-09-01 → 2028-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10914914

## Citation

> US National Institutes of Health, RePORTER application 10914914, Project 1: KSHV reprograms replication and metabolic activities in hypoxia (5P01CA281867-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10914914. Licensed CC0.

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