JAUNDICE NEXT: A diagnostic tool for cholestatic liver disease.

DESCRIPTION (provided by applicant): Cholestasis (or pathologic jaundice) is the most common sign of liver disease in children and often results from mutations in genes that play a critical role in bile formation and transport. In adults, mutations in thesame genes also cause chronic cholestasis that may manifest as non-specific portal fibrosis, intrahepatic cholestasis of pregnancy, and low-phospholipid-associated cholelithiasis. Using recent knowledge of the genetic basis of these disorders, we developeda high-throughput gene chip that identifies mutations in the genes that cause the most common forms of inherited syndromes of chronic cholestasis. The chip is now a clinical test that is increasingly used by physicians as part of diagnostic algorithms insymptomatic patients. Although the chip remarkably facilitates specific diagnoses, its accuracy is limited by the inability to reliably detect insertions or deletions, whih account for 10-20% of the disease-causing mutations, thus decreasing the sensitivity of the chip. In this application, we propose to solve this technological gap by developing the next version of a mutation-screening tool that we are naming JAUNDICENEXT . This tool customizes NextGen sequencing to accurately survey the nucleotide composition of target genes to diagnose disease-causing mutations. Our Specific Aim is to determine the accuracy of JAUNDICENEXT to sequence the target genes. To test Hypothesis-1 that JAUNDICENEXT sequences the target genes at gt99% accuracy, we willcompare the nucleotide sequence produced by the JaundiceNext with the sequence produced by standard sequencing methodology. To test Hypothesis-2 that JAUNDICENEXT identifies insertion and deletion mutations in patients with cholestasis, we will examinewhether the sequence output produced by JAUNDICENEXT detects insertions, deletions, or indels in DNA known to carry these mutations. Collectively, these experiments will validate the technical merits of JAUNDICENEXT and position us for future experimentsto further bench-test the assay and develop an automated detection algorithm to in a future Phase-II application. PUBLIC HEALTH RELEVANCE: Jaundice is a clinical sign that is common to several types of chronic liver diseases in children and adults. In a previous STTR, we developed a gene chip to identify mutations in the genes that cause the most common forms of inherited syndromes of persistent jaundice (also known as cholestasis). The chip is now a clinical test that is increasingly used by physicians as part of diagnostic algorithms in symptomatic patients. Although the chip remarkably facilitates specific diagnoses, its accuracy is limited by the inabiliy to reliably detect insertions or deletions, which account for 10-20% of the disease-causingmutations, thus decreasing the sensitivity of the chip. We propose to solve this technological gap by developing the next version of a mutation-screening tool that we are naming JAUNDICENEXT . This new tool customizes NextGen, a powerful sequencing technology, to accurately detect the wider spectrum of mutations in four genes simultaneously. These studies will improve the sensitivity and accuracy of a non-invasive diagnostic tool that significantly improves the diagnostic algorithm in patients with liver disease.