Human papillomavirus mRNA detection for assessing cervical cancer risk in resource-limited settings

ABSTRACT Cervical cancer is highly preventable with early detection and treatment, but due to lack of accessible early screening options, incidence and mortality are still very high in low-resource settings (LRS). High-risk human papillomavirus (hrHPV) is responsible for most cervical cancer cases, and the WHO recommends testing for

hrHPV and preemptively treating all patients who are positive as the primary prevention strategy in LRS. Nearly all available tests look for hrHPV DNA, which can result in overtreatment because it cannot differentiate infections that will clear naturally. Detectable hrHPV mRNA, however, is strongly associated with higher grade cervical

precancers, making it a more specific biomarker for cervical cancer risk. This proposal aims to address the need for more specific early cervical cancer detection technology by developing novel in vitro and in vivo methods for detecting hrHPV mRNA. In the F99 phase of this proposal, I will develop and pilot a sample-to-answer mRNA

test for HPV16 and HPV18. I will first develop a minimally instrumented method for preparing mRNA for detection from both provider and self-collected cervical samples. I will then amplify this mRNA using an isothermal assay that produces a real-time fluorescent signal, which I will read using a low-cost fluorimeter. I will integrate the

individual assay components into a workflow with minimal user steps, making a test that is deployable to LRS. I will work with my sponsor, Dr. Rebecca Richards-Kortum, an established expert in the field of point-of-care cancer detection technologies to develop this test. I will then work with my co-sponsor, Dr. Kathleen Schmeler,

the VP of Global Oncology at the UT MD Anderson Cancer Center with extensive experience translating cancer detection technologies to LRS to pilot this test with clinical samples in both Houston and Brazil. This training plan will help me gain experience with technology development, scientific communication, and clinical collaborations

both locally and globally, and is enhanced by the location of my training at Rice University in the Texas Medical Center, where I will have access to world-class equipment, resources, and clinical collaborators. In the K00 phase of this proposal, I will develop a technique for detecting hrHPV mRNA in vivo. This will permit real-time

early cervical cancer detection and monitoring, which can help assess disease progression and inform treatment faster than repeated in vitro sampling and testing. In the proposed project, I will use a combination of sequence- specific fluorescent mRNA labeling, aptamer-mediated label delivery, and high-resolution fluorescence imaging

to detect hrHPV mRNA sensitively and specifically in vivo. I will seek a mentor with experience in biomarker label design and delivery at an institution with robust imaging resources and clinical collaborations. The proposed work will prepare me for an academic career as a cancer researcher dedicated to improving global access to

early cancer detection. In addition to technical experience, I will develop my scientific communication skills, build a network of collaborators, and mentor the next generation of cancer researchers, laying the foundation for a career developing and deploying novel cancer prevention technologies with clinical impact around the world.