Combinatorial CRISPR-Cas9 screens for de novo mapping of genetic interactions.

We developed a systematic approach to map human genetic networks by combinatorial CRISPR-Cas9 perturbations coupled to robust analysis of growth kinetics. We targeted all pairs of 73 cancer genes with dual guide RNAs in three cell lines, comprising 141,912 tests of interaction. Numerous therapeutically relevant interactions were identified, and these patterns replicated with combinatorial drugs at 75% precision. From these results, we anticipate that cellular context will be critical to synthetic-lethal therapies

Age- and sex-related changes in fasting plasma glucose and lipoprotein in cynomolgus monkeys

BACKGROUND: The age-related dysfunction of glucose and lipid metabolism has a long-standing relationship with cardiovascular and neurodegenerative disease. However, the effects of metabolic dysfunction on men and women are different. Reasons for these sex differences remains unclear. Cynomolgus monkeys have been used, in the past, for the study of human metabolic diseases due to their biologically proximity to humans. Nevertheless, few studies to date have focused on both age- and sex-related differences in glucose and lipid metabolism. The present study was designed to specifically address these questions by using a large cohort of cynomolgus monkeys (N = 1,399) including 433 males and 966 females with ages ranging 4 to 24 years old. METHODS: Fasting plasma glucose (FPG) and lipid parameters including total cholesterol (T-Cho), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured. All these parameters were compared between ages and sexes. RESULTS: Among the entire cohort, age was strongly correlated with levels of FPG, TG and HDL. Consequently, sex-related analysis revealed that females had significantly higher average levels of FPG, T-Cho, TG, HDL-C and LDL-C than their male counterparts. In addition, more female (28.5 %) than male (16 %) monkeys qualified for impaired fasting plasma glucose (IFPG). In those IFPG animals, sex-related differences were also detected i.e. females had significantly increased levels of T-Cho, TG and LDL-C. CONCLUSIONS: The result, for the first time, demonstrated the similarities and differences in detail between male and female cynomolgus monkeys in relationship to age-related glucose and lipoprotein metabolisms, and differences under various physiological conditions. The detailed glucose and lipoprotein profiling should provide additional and important insights for prediabetic conditions. Cynomolgus monkeys appear to be an excellent model for translational research of diabetes and for novel therapeutic strategies testing to overt diabetes

Human malarial disease: a consequence of inflammatory cytokine release

Malaria causes an acute systemic human disease that bears many similarities, both clinically and mechanistically, to those caused by bacteria, rickettsia, and viruses. Over the past few decades, a literature has emerged that argues for most of the pathology seen in all of these infectious diseases being explained by activation of the inflammatory system, with the balance between the pro and anti-inflammatory cytokines being tipped towards the onset of systemic inflammation. Although not often expressed in energy terms, there is, when reduced to biochemical essentials, wide agreement that infection with falciparum malaria is often fatal because mitochondria are unable to generate enough ATP to maintain normal cellular function. Most, however, would contend that this largely occurs because sequestered parasitized red cells prevent sufficient oxygen getting to where it is needed. This review considers the evidence that an equally or more important way ATP deficency arises in malaria, as well as these other infectious diseases, is an inability of mitochondria, through the effects of inflammatory cytokines on their function, to utilise available oxygen. This activity of these cytokines, plus their capacity to control the pathways through which oxygen supply to mitochondria are restricted (particularly through directing sequestration and driving anaemia), combine to make falciparum malaria primarily an inflammatory cytokine-driven disease