Gregory C. Loney, Ph.D. - US grants

Click Diagnostics has developed a new low-cost, disposable, PCR-based diagnostic test for multiplexed detection of the sexually transmitted infections (STI) chlamydia, gonorrhea, and trichomoniasis. The Centers for Disease Control (CDC) estimates that nearly 20 million new STIs occur every year and account for almost $16 billion in health care costs. Current STI diagnostic testing is primarily done in centralized labs with an average sample-to-result-to-patient timeline of 3 to 5 days. During this time, as many as 30% of STI-positive patients are lost to care and continue to spread the STI. Many others are given inappropriate drugs to avoid the patient leaving without treatment, despite not having test results. Although point-of-care (POC) nucleic acid amplification based instruments are available, they can perform only one test at a time, limiting their usefulness in a busy clinic. To ensure patients receive the appropriate antibiotic, a highly sensitive and specific, affordable, POC diagnostic that provides rapid results is urgently needed. The Click patient-side, single-use disposable, polymerase chain reaction (PCR) diagnostic test, with near 100% accuracy, would allow physicians to ensure no patient with an STI leaves the clinic untreated. Click has developed prototype units, and successfully validated them using clinical vaginal swab samples from patients. In this project, we will refine the device to enable mass production, and optimize the device in preparation for clinical trials, 510(k) regulatory clearance, and a CLIA waiver. This will require product refinements for manufacturability, performing preanalytical and preclinical validation, and usability studies. The goal of Aim 1 is to optimize the pre-production unit for manufacturability and clinical testing. This will require using low cost fabrication techniques, minimizing the part count to reduce unit complexity, and reducing device size and weight. These modifications enable manufacturing readiness to produce units for preanalytical and preclinical testing in Aim 2. Preliminary analytical studies will evaluate the device limit of detection and demonstrate the device results are reproducible and specific. Click has partnered with Planned Parenthood, University of Alabama, and Johns Hopkins University for preclinical testing to validate the Click diagnostic test. Click will also verify test results against a gold standard PCR instrument by Roche. These tests will assess readiness for clinical testing as required for 510(k) regulatory submission. All tests will be done under strict quality control and document control with Click?s advanced quality management system. Lastly, in Aim 3 we will conduct a usability study to ensure that untrained operators can successfully operate the device, as would be required for a CLIA waiver. A CLIA waiver is critical for Click to market its product to physician offices, and pharmacies. Based on user feedback, Click will make any necessary modifications to the device or written instructions to meet CLIA waiver requirements. Successful completion of this work will yield a production version of a rapid, patient-side nucleic acid-based diagnostic device ready for clinical testing.

Project Summary/Abstract Concurrent nicotine use is highly implicated in the propensity to misuse prescribed opiates, future development of opiate dependence, and susceptibility to accidental opiate overdose. Approximately 83-95% of patients involved in opioid treatment programs are tobacco users. As such, there exists a clear public health mandate for elucidation of mechanisms contributing to opiate addiction liability resultant from opiate and nicotine polysubstance use. The anterior insular cortex is heavily involved in processing the interoceptive stimulus properties of commonly abused drugs and insular dysfunction is implicated in the development and maintenance of substance use disorders. The role of the insular cortex in opiate addiction, and furthermore, how nicotine may play a role, is currently underexplored. There is emerging evidence that nicotine interferes with insular function, specifically through depressing synaptic potentiation and inhibiting the output of the insular cortex. Preliminary data demonstrate that nicotine delivered to the anterior insular cortex interferes with the processing of the stimulus properties of opiates in a quantitative manner, an effect remarkably similar to insular chemogenetic inactivation. Specifically, subjects either administered nicotine or undergoing chemogenetic inactivation of the insular cortex behave as if they have received a lower dose of morphine relative to controls in paradigms examining both the reinforcing (conditioned place preference; CPP) and aversive (conditioned taste avoidance; CTA) stimulus properties of opiates. As a result, tobacco smokers may require more and stronger doses of opiates in order to achieve the desired effects. This reduced sensitivity to the stimulus properties of opiates due to nicotine co-use may facilitate the development of opiate dependence. The goal of this project is to elucidate the mechanisms and neural circuits through which insular cortical nicotinic activity contributes to enhanced opiate addiction liability. These experiments will employ a combination of psychophysical behavioral measures, site-specific behavioral pharmacology, circuit-specific chemogenetic manipulation, and in vivo electrophysiology in order to fully characterize the mechanisms through which insular cortical nicotinic activity may contribute to the development of opiate substance use disorders. Furthermore, through completion of these proposed studies, the applicant will be competently trained in new technical skills and perspectives, ultimately facilitating a successful transition to a principal investigator conducting programmatic research on the comorbidity of nicotine and opiate abuse.