# AND-gated Synthetic Biomarkers for Early Detection of Liver Metastasis

> **NIH NIH U01** · GEORGIA INSTITUTE OF TECHNOLOGY · 2021 · $599,590

## Abstract

PROJECT SUMMARY
Advances in synthetic biology will play a fundamental role in shaping the future of cancer diagnostics toward
earlier and more specific detection of disease. For example, whole-cell biosensors such as bacteria have been
genetically engineered to perform complex functions such as signal amplification to detect clinically relevant
biomarkers in human urine and serum. In mammalian cells, sense-and-respond components that employ
Boolean logic have been demonstrated for multiplexed control of engineered T cell therapies, thereby increasing
the specificity of tumor sensing and reducing systemic toxicity. These advances highlight the promise of synthetic
biology when applied to cancer, yet the vast majority of these strategies rely on genetic circuits and make use of
non-mammalian protein components. Such circuits are complex and raise safety and immunogenicity concerns
for regulatory approval, especially in the context of in vivo early cancer detection where repeated administrations
of biosensors are likely needed to monitor for nascent disease. This proposal seeks to develop a new class of
diagnostics called AND-gated synthetic biomarkers for early detection of cancer metastasis. Synthetic
biomarkers are an emerging class of activatable biological sensors that are designed to be administered
systemically, query sites of early disease, and harness tumor-dependent activation mechanisms, such as
dysregulated protease activity, to drive production of a reporter. These reporters can then be detected
noninvasively from blood, urine, or other bodily fluid samples. Proteases play key biological roles across the
major hallmarks of metastasis and are particularly potent molecular amplifiers by catalyzing the irreversible
hydrolysis of peptide bonds, allowing a single protease to turnover thousands of substrates. AND-gated synthetic
biomarkers will be applied for early detection of colorectal cancer (CRC) liver metastasis. Although the liver is a
common site for metastatic spread from primary CRC, regional resection of liver-isolated metastases can lead
to potentially curative results. Yet early detection of CRC liver metastases at a size when they are most
responsive to therapy (1–2 mm) remains challenging by radiographic imaging such as CT and FDA-approved
blood test such as the carcinoembryonic antigen (CEA) test. To design AND-gated synthetic biomarkers for CRC
liver metastasis, pairwise combinations of proteases will be selected based on differential RNA expression in
CRC liver metastases compared to healthy liver tissue. Multivariate mathematical models will be developed to
understand how design parameters enhance specificity and cooperativity compared to experimental results.
Syngeneic and xenograft models of CRC liver metastasis will be used for preclinical validation studies to allow
benchmarking against CT and CEA. This proposal will lay the groundwork for earlier detection of cancer
metastasis by programmable synthetic biomarkers.

## Key facts

- **NIH application ID:** 10330265
- **Project number:** 1U01CA265711-01
- **Recipient organization:** GEORGIA INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** Gabriel A Kwong
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $599,590
- **Award type:** 1
- **Project period:** 2021-09-23 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10330265

## Citation

> US National Institutes of Health, RePORTER application 10330265, AND-gated Synthetic Biomarkers for Early Detection of Liver Metastasis (1U01CA265711-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10330265. Licensed CC0.

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