Truncation of Pik3r1 causes severe insulin resistance uncoupled from obesity and dyslipidaemia by increased energy expenditure.

OBJECTIVE: Insulin signalling via phosphoinositide 3-kinase (PI3K) requires PIK3R1-encoded regulatory subunits. C-terminal PIK3R1 mutations cause SHORT syndrome, as well as lipodystrophy and insulin resistance (IR), surprisingly without fatty liver or metabolic dyslipidaemia. We sought to investigate this discordance. METHODS: The human pathogenic Pik3r1 Y657∗ mutation was knocked into mice by homologous recombination. Growth, body composition, bioenergetic and metabolic profiles were investigated on chow and high-fat diet (HFD). We examined adipose and liver histology, and assessed liver responses to fasting and refeeding transcriptomically. RESULTS: Like humans with SHORT syndrome, Pik3r1WT/Y657∗ mice were small with severe IR, and adipose expansion on HFD was markedly reduced. Also as in humans, plasma lipid concentrations were low, and insulin-stimulated hepatic lipogenesis was not increased despite hyperinsulinemia. At odds with lipodystrophy, however, no adipocyte hypertrophy nor adipose inflammation was found. Liver lipogenic gene expression was not significantly altered, and unbiased transcriptomics showed only minor changes, including evidence of reduced endoplasmic reticulum stress in the fed state and diminished Rictor-dependent transcription on fasting. Increased energy expenditure, which was not explained by hyperglycaemia nor intestinal malabsorption, provided an alternative explanation for the uncoupling of IR from dyslipidaemia. CONCLUSIONS: Pik3r1 dysfunction in mice phenocopies the IR and reduced adiposity without lipotoxicity of human SHORT syndrome. Decreased adiposity may not reflect bona fide lipodystrophy, but rather, increased energy expenditure, and we suggest that further study of brown adipose tissue in both humans and mice is warranted

T cell phenotyping of a mouse model of Activated PI3Kdelta syndrome

Activated PI3Kdelta Syndrome (APDS) is immunodeficiency caused by a heterozygous gain-of-function mutation (E1021K) in the PIK3CD gene, encoding for the p110delta catalytic subunit of phosphoinositide 3-kinase (PI3K). APDS patients are lymphopenic, suffer from sinopulmonary infections and from increased susceptibility to bacterial and herpes group virus infections. Following T cell receptor (TCR) stimulation, T cells from these patients undergo increased activation induced cell death, which can be reversed by selective PI3Kdelta inhibitors. I used a new conditional knock-in mouse (T-p110delta E1020K) in order to investigate the effect of hyperactive p110delta on T cell function. Hyperactivation of p110delta led to increased PIP3 and pAKT levels following TCR stimulation that was reduced using a selective p110delta inhibitor. Following in vitro TCR stimulation, T cells proliferated normally but showed increased apoptosis that was reversed by a p110delta specific inhibitor. Despite enhanced apoptosis, CD8+ T cells displayed enhanced activation that was associated with increased levels of cytokines and granzyme B. CD4+ T cells with hyperactive p110delta produced increased Th1, Th2, Th17 and Tfh cytokines but showed reduced Treg differentiation in vitro. Conditional T-p110delta E1020K mice have reduced Tregs in the thymus but increased peripheral Tregs. These mice also have increased numbers of T follicular helper cells and germinal centre (GC) B cells upon immunisation with a T cell dependent antigen (NP-KLH). Reduced antigen specific IgG1+ cells within GC B cells was detected in mice harbouring hyperactive p110delta mutation in B cells, implying that the antibody deficiencies observed in APDS patients is due to an intrinsic defect within B cells rather than limited help from T cells. T-p110delta E1020K mice mounted normal primary responses to acute infections .However adoptive transfer of ovalbumin-specific T cell receptor transgenic CD8 T cells (OT1) revealed an intrinsic defect in the primary expansion of CD8+ T cells with hyperactive p110delta. This defect in primary expansion was found to be rescued in the presence of wild type OT1 cells. Following infection with acute pathogens, CD8+ T cells with hyperactive p110delta displayed normal to increased effector function with phenotypically reduced memory cells as indicated by reduced memory precursor effectors cells (MPECs). In contrast, following chronic infection, T-p110delta E1020K displayed increased signs of T cell exhaustion that is also characteristic of APDS patients as they suffer from chronic herpes virus infections. This set of work therefore shows that the mouse model recapitulates key aspects of APDS patients and give insights into the role of p110delta signaling in different T cell subsets influenced by hyperactive p110delta activity. Further, in depth analysis of proteomics and gene array data of in vitro stimulated T cells generated during this study, can provide a better understanding of the mechanisms behind the T cell phenotype observed

Truncation of Pik3r1 causes severe insulin resistance uncoupled from obesity and dyslipidaemia by increased energy expenditure.

OBJECTIVE: Insulin signalling via phosphoinositide 3-kinase (PI3K) requires PIK3R1-encoded regulatory subunits. C-terminal PIK3R1 mutations cause SHORT syndrome, as well as lipodystrophy and insulin resistance (IR), surprisingly without fatty liver or metabolic dyslipidaemia. We sought to investigate this discordance. METHODS: The human pathogenic Pik3r1 Y657∗ mutation was knocked into mice by homologous recombination. Growth, body composition, bioenergetic and metabolic profiles were investigated on chow and high-fat diet (HFD). We examined adipose and liver histology, and assessed liver responses to fasting and refeeding transcriptomically. RESULTS: Like humans with SHORT syndrome, Pik3r1WT/Y657∗ mice were small with severe IR, and adipose expansion on HFD was markedly reduced. Also as in humans, plasma lipid concentrations were low, and insulin-stimulated hepatic lipogenesis was not increased despite hyperinsulinemia. At odds with lipodystrophy, however, no adipocyte hypertrophy nor adipose inflammation was found. Liver lipogenic gene expression was not significantly altered, and unbiased transcriptomics showed only minor changes, including evidence of reduced endoplasmic reticulum stress in the fed state and diminished Rictor-dependent transcription on fasting. Increased energy expenditure, which was not explained by hyperglycaemia nor intestinal malabsorption, provided an alternative explanation for the uncoupling of IR from dyslipidaemia. CONCLUSIONS: Pik3r1 dysfunction in mice phenocopies the IR and reduced adiposity without lipotoxicity of human SHORT syndrome. Decreased adiposity may not reflect bona fide lipodystrophy, but rather, increased energy expenditure, and we suggest that further study of brown adipose tissue in both humans and mice is warranted

PI3Kδ hyper-activation promotes development of B cells that exacerbate Streptococcus pneumoniae infection in an antibody-independent manner.

Streptococcus pneumoniae is a major cause of pneumonia and a leading cause of death world-wide. Antibody-mediated immune responses can confer protection against repeated exposure to S. pneumoniae, yet vaccines offer only partial protection. Patients with Activated PI3Kδ Syndrome (APDS) are highly susceptible to S. pneumoniae. We generated a conditional knock-in mouse model of this disease and identify a CD19+B220- B cell subset that is induced by PI3Kδ signaling, resides in the lungs, and is correlated with increased susceptibility to S. pneumoniae during early phases of infection via an antibody-independent mechanism. We show that an inhaled PI3Kδ inhibitor improves survival rates following S. pneumoniae infection in wild-type mice and in mice with activated PI3Kδ. These results suggest that a subset of B cells in the lung can promote the severity of S. pneumoniae infection, representing a potential therapeutic target.Wellcome, MRC, BBSR