Principal Investigator (PI)
Biomedical Primate Research Centre
The Foundation is providing £141,552 in support.
Anke Harupa - Lars Vermaat
We envisage GSK’s contribution in the field of hepatocyte culture, which will be of great value to the project. The Open Lab scientist will perform part of the hepatocyte culture optimization at GSK facilities, GSK will provide consumables for these periods in kind as well as mentoring from experienced researchers.
The BPRC P. cynomolgi in vitro liver stage drug assay, enabling drug screening on developing- and dormant liver stages (hypnozoites), may be biased towards prophylactic compound activity, rather than radical curative activity. The assay is performed with primary rhesus monkey hepatocytes and drug exposure from day 0 to 6 days post sporozoite infection. We are currently developing an assay that may be more predictive for radical curative activity, by applying compound exposure from day 5 to 9 post infection. Primaquine (PQ), the only positive control with known radical curative activity in vivo, has variable activity in the standard assay and no activity in the radical cure type assay. Thus, we are lacking a positive control in the assay. As PQ needs to be metabolized to display anti-hypnozoite activity, it is likely that declining metabolic activity over time of the primary hepatocytes causes this variable activity in the standard assay and absence of activity in the radical cure type assay. We plan to address this by first fully characterizing hepatocytes during culture and in the liver prior to isolation. This includes typical hepatocyte markers as well as the metabolism capacity. We will then apply alternative culture methods and monitor PQ metabolism over time. Finally, we will monitor parasite invasion and growth and PQ anti-parasite activity over time, and adapt culture conditions further to arrive at an optimal radical cure type assay. Once the radical cure assay is optimized, a set of compounds will be screened, using PQ as the positive control, aiming to identify novel compounds with radical cure activity.