Hypoxia promotes liver stage malaria infection in primary human hepatocytes in vitro

Abstract

Homeostasis of mammalian cell function strictly depends on balancing oxygen exposure to maintain energy metabolism without producing excessive reactive oxygen species. In vivo, cells in different tissues are exposed to a wide range of oxygen concentrations, and yet in vitro models almost exclusively expose cultured cells to higher, atmospheric oxygen levels. Existing models of liver-stage malaria that utilize primary human hepatocytes typically exhibit low in vitro infection efficiencies, possibly due to missing microenvironmental support signals. One cue that could influence the infection capacity of cultured human hepatocytes is the dissolved oxygen concentration. We developed a microscale human liver platform comprised of precisely patterned primary human hepatocytes and nonparenchymal cells to model liver-stage malaria, but the oxygen concentrations are typically higher in the in vitro liver platform than anywhere along the hepatic sinusoid. Indeed, we observed that liver-stage Plasmodium parasite development in vivo correlates with hepatic sinusoidal oxygen gradients. Therefore, we hypothesized that in vitro liver-stage malaria infection efficiencies might improve under hypoxia. Using the infection of micropatterned co-cultures with Plasmodium berghei, Plasmodium yoelii or Plasmodium falciparum as a model, we observed that ambient hypoxia resulted in increased survival of exo-erythrocytic forms (EEFs) in hepatocytes and improved parasite development in a subset of surviving EEFs, based on EEF size. Further, the effective cell surface oxygen tensions (pO2) experienced by the hepatocytes, as predicted by a mathematical model, were systematically perturbed by varying culture parameters such as hepatocyte density and height of the medium, uncovering an optimal cell surface pO2 to maximize the number of mature EEFs. Initial mechanistic experiments revealed that treatment of primary human hepatocytes with the hypoxia mimetic, cobalt(II) chloride, as well as a HIF-1α activator, dimethyloxalylglycine, also enhance P. berghei infection, suggesting that the effect of hypoxia on infection is mediated in part by host-dependent HIF-1α mechanisms.

Transcriptome analysis illuminates the nature of the intracellular interaction in a vertebrate-algal symbiosis

“…Plasmodium falciparum exhibits increased infection and growth at low partial pressures of oxygen (Ng et al, 2014; Scheibel et al, 1979). Moreover, the observed cell stress response of the alga is reminiscent of that experienced by intracellular apicomplexan parasites (Bosch et al, 2015).…”

Characteristic MicroRNA Expression Induced by δ-Opioid Receptor Activation in the Rat Liver Under Prolonged Hypoxia

“…Hypoxia is associated with various clinical conditions including liver disorders such as fatty liver disease [29], liver-stage malaria [30], and hepatocellular carcinoma [31]. Hepatacellular cells need to activate specific molecular programs to overcome hypoxic challenges in these conditions [32-34].…”

Micropatterned coculture of primary human hepatocytes and supportive cells for the study of hepatotropic pathogens

“…MPCCs have been used to study basic mechanisms underlying hepatocyte-stromal co-cultures, drug metabolism and toxicity 22–26 , and most recently, the life cycle of HCV 15 and human Plasmodium 16,27 pathogens. As MPCCs recapitulate the life cycle of these pathogens, they open the door to studying infection pathophysiology and drug response, as we will discuss below.…”

“…For instance, MPCCs have been shown to be ~70-75% predictive of clinical outcomes for drug metabolite and drug-induced liver toxicity profiling as opposed to <50% sensitivity in standard culture systems 22 . A more recent application of MPCCs has been the study of hepatotropic pathogens 15,16,27,28 . While the present report focuses on the use of MPCCs to study HBV, HCV and Plasmodium infection (Figure 1), this coculture platform can also be utilized to study hepatotropic pathogens broadly.…”