Together with the School of Medicine this grant mechanism funds innovative, and currently unfunded, translational science projects that feature a collaboration of at least two investigators who are project Co-PIs. For this call only projects were focused on infectious diseases.
Hye-won Shin, PhD, Department of Pediatrics
Don Blake, PhD, Department of Chemistry
Non-Invasive Diagnosis of Invasive Pulmonary Aspergillosis Using Exhaled Breath Gases
Weian Zhao, PhD; Department of
Ellena Peterson, PhD; Department of Pathology
Detection of single bacteria in blood infection in minutes
Blood infection caused by bacteria is a major health problem which annually affects over 18 million people worldwide and 700,000 in the U.S. just from sepsis alone, with a mortality rate of approximately 30-40%. A major challenge in blood infection treatment is diagnosis: by the time doctors identify the bacteria species to refine treatment, it can be too late. Indeed, the current gold standard for bacteria detection, blood culture, takes days to get a result while other molecular diagnosis methods such as PCR are often not sensitive enough to detect bacteria that occur at low concentrations in blood (1-100 CFU/mL). We propose a device that rapidly and inexpensively counts bacteria in patient’s blood at singlecell sensitivity without any sample preparation within several minutes. The device integrates a droplet-based microfluidics system with functional DNAzyme sensors that are identified via in vitro selection to specifically report target bacteria with a rapid, real-time fluorescence signal. The confinement of bacteria samples along with DNAzyme sensors into millions of picoliter droplets significantly increases the concentration of released target molecules, facilitating high throughput detection of single bacterial cells in complex mixtures such as blood. In Aim 1, we will integrate DNAzyme sensors that detect E. coli with droplet microfluidics to test and optimize the single-cell detection of bacteria in both buffer and blood. In Aim 2, we will validate the reliability of our device, including both sensitivity and specificity, in clinical specimens by correlating the results of our device with blood culture and PCR. This study will lead to a new paradigm approach for rapid detection of bacteria in blood infections, providing clinicians with needed information to improve treatments and reduce the mortality associated with bacterial blood infections.
Lan Huang, PhD; Department of
Physiology and Biophysics
Luis de la Maza, MD; Department Pathology and Laboratory Medicine
Defining the Structure of Native C.trachomatis MOMP for Vaccine Development
Chlamydia trachomatis is the main cause of preventable blindness, and the most common sexually transmitted bacterial disease worldwide. Vaccination is the only viable approach to the control and eradication of Chlamydia. The chlamydial major outer membrane protein (MOMP) is the most promising vaccine candidate. Using the native MOMP (nMOMP) as the antigen, significant protection was observed against genital, ocular and respiratory challenges. In contrast, a recombinant preparation of the MOMP (rMOMP), that lacks the correct structural conformation, is a significantly less effective vaccine than the nMOMP. Our assumption is that the rMOMP construct did not present epitopes to the host immune system in the 3-dimensional conformation in which the epitopes appear on intact bacteria. The structural conformation of the nMOMP is unknown. Our goal is to determine the structure of the C trachomatis MOMP using novel cross-linking mass spectrometry technologies for use in the development of a vaccine. These studies will provide much needed structural data to formulate a vaccine to protect against chlamydial infections.
This is a collaborative project between Drs. Lan Huang and Luis de la Maza. Dr. Huang is one of the leaders in the field of proteomics who has developed and employed novel crosslinking/mass spectrometry strategies to study protein interactions and structures of protein complexes. Dr. de la Maza is the leader in the field of Chlamydia vaccinology and has been studying the native major outer membrane protein (nMOMP) from C. trachomatis for more than 20 years, which is the most promising vaccine candidate. The two PIs form an ideal team to address a major knowledge gap in Chlamydia research by determining the structural conformation of the nMOMP and its protective epitopes for designing a more effective Chlamydia vaccine. The two PIs are among the leaders in their fields and there is no other combination of PIs better qualified to develop a new Chlamydia vaccine. This collaboration will lead to a highly significant scientific and translational breakthrough and benefit the health of tens of millions of people.