Brain Distribution and Efficacy of the Brain Penetrant PI3K Inhibitor GDC-0084 in Orthotopic Mouse Models of Human Glioblastoma.

Glioblastoma multiforme (GBM) is the most common primary brain tumor in adults. Limited treatment options have only marginally impacted patient survival over the past decades. The phophatidylinositol 3-kinase (PI3K) pathway, frequently altered in GBM, represents a potential target for the treatment of this glioma. 5-(6,6-Dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[4,3-e]purin-2-yl)pyrimidin-2-amine (GDC-0084) is a PI3K inhibitor that was specifically optimized to cross the blood-brain barrier. The goals of our studies were to characterize the brain distribution, pharmacodynamic (PD) effect, and efficacy of GDC-0084 in orthotopic xenograft models of GBM. GDC-0084 was tested in vitro to assess its sensitivity to the efflux transporters P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) and in vivo in mice to evaluate its effects on the PI3K pathway in intact brain. Mice bearing U87 or GS2 intracranial tumors were treated with GDC-0084 to assess its brain distribution by matrix-assisted laser desorption ionization (MALDI) imaging and measure its PD effects and efficacy in GBM orthotopic models. Studies in transfected cells indicated that GDC-0084 was not a substrate of P-gp or BCRP. GDC-0084 markedly inhibited the PI3K pathway in mouse brain, causing up to 90% suppression of the pAkt signal. MALDI imaging showed GDC-0084 distributed evenly in brain and intracranial U87 and GS2 tumors. GDC-0084 achieved significant tumor growth inhibition of 70% and 40% against the U87 and GS2 orthotopic models, respectively. GDC-0084 distribution throughout the brain and intracranial tumors led to potent inhibition of the PI3K pathway. Its efficacy in orthotopic models of GBM suggests that it could be effective in the treatment of GBM. GDC-0084 is currently in phase I clinical trials.

Effects of race on upper airway dynamic function during sleep in children.

STUDY OBJECTIVE: Studies in adults and children have shown that African American race is a risk factor for the obstructive sleep apnea syndrome (OSAS). Therefore, we hypothesized that non-obese, non-snoring African American children would have a more collapsible upper airway during sleep than age-, gender-, and size-matched Caucasians. DESIGN: Upper airway dynamic function was measured during sleep in normal African American and Caucasian children. SETTING: Sleep laboratory. PATIENTS OR PARTICIPANTS: 56 normal children between the ages of 8-18 years. INTERVENTIONS: Pressure-flow relationships were measured during NREM sleep. Nasal pressure was decreased to subatmospheric levels, using previously described techniques that resulted in an activated and relatively hypotonic upper airway. MEASUREMENTS AND RESULTS: The activated and hypotonic critical pressures (Pcrit) were -25 (-25, -3) (median, range) and -19 (-25, -3) for African Americans, and -25 (-25, -4) and -25 (-25.0, -4) cm H(2)O, respectively, for Caucasians. The slopes of the pressure-flow response (SPF) under activated and hypotonic conditions for African Americans were 10 (-9, 46) and 13 (-20, 46), and for Caucasians 9 (-9, 64) and 8 (-5, 54) mL/s/cm H(2)O, respectively. There were no significant differences between groups for Pcrit or SPF under either activated or hypotonic conditions. CONCLUSION: Upper airway collapsibility was similar in asymptomatic, non-obese African American and Caucasian children. Differences in upper airway characteristics and neuromotor function cannot explain the increased prevalence of OSAS in African American children.

Upper airway collapsibility during REM sleep in children with the obstructive sleep apnea syndrome.

STUDY OBJECTIVES: In children, most obstructive events occur during rapid eye movement (REM) sleep. We hypothesized that children with the obstructive sleep apnea syndrome (OSAS), in contrast to age-matched control subjects, would not maintain airflow in the face of an upper airway inspiratory pressure drop during REM sleep. DESIGN: During slow wave sleep (SWS) and REM sleep, we measured airflow, inspiratory time, inspiratory time/total respiratory cycle time, respiratory rate, tidal volume, and minute ventilation at a holding pressure at which flow limitation occurred and at 5 cm H2O below the holding pressure in children with OSAS and in control subjects. SETTING: Sleep laboratory. PARTICIPANTS: Fourteen children with OSAS and 23 normal control subjects. RESULTS: In both sleep states, control subjects were able to maintain airflow, whereas subjects with OSAS preserved airflow in SWS but had a significant decrease in airflow during REM sleep (change in airflow of 18.58 +/- 12.41 mL/s for control subjects vs -44.33 +/- 14.09 mL/s for children with OSAS, P = 0.002). Although tidal volume decreased, patients with OSAS were able to maintain minute ventilation by increasing the respiratory rate and also had an increase in inspiratory time and inspiratory time per total respiratory cycle time CONCLUSION: Children with OSAS do not maintain airflow in the face of upper-airway inspiratory-pressure drops during REM sleep, indicating a more collapsible upper airway, compared with that of control subjects during REM sleep. However, compensatory mechanisms exist to maintain minute ventilation. Local reflexes, central control mechanisms, or both reflexes and control mechanisms need to be further explored to better understand the pathophysiology of this abnormality and the compensation mechanism.

Polysomnographic values in children undergoing puberty: pediatric vs. adult respiratory rules in adolescents.

STUDY OBJECTIVES: Polysomnographic respiratory events in children should be scored using pediatric respiratory rules. However, due to a lack of data on adolescents, recently revised rules allow children aged 13-18 years to be scored by adult or pediatric criteria. To clarify which criteria to use, we describe the evolution of respiratory events with Tanner stage, and we compare events in children aged 13-18 years with the new American Academy of Sleep Medicine adult and pediatric respiratory rules. DESIGN: Cross-sectional SETTING: Academic hospital PARTICIPANTS: Healthy subjects aged 8-18 years recruited for research purposes. INTERVENTIONS: Physical examination to determine Tanner stage, overnight polysomnogram, and determination of sex hormones. RESULTS: Sixty-eight subjects (Tanner 1-5) were studied, mean age [SD] = 13 +/- 3 years, median apnea hypopnea index (AHI)= 0.1 (range: 0-1.2)/h. The median percentages of total sleep time (TST) with SpO2 < 92% were 0.1 (0-4.2)%, and with end-tidal CO2 > 50 torr was 0.1 (0-88.6)%. Thirty-two subjects were aged 13-18 years, (Tanner 3-5). The difference between AHI scored by pediatric (median = 0 [0-0.9]/h) and adult (median = 0 [0 - 0.5]/h) criteria was statistically significant (P = 0.043), but not clinically relevant. CONCLUSIONS: Respiratory events in normal children aged 8-18 years are rare and unrelated to Tanner stage. Adult or pediatric respiratory rules can be used for scoring polysomnograms in asymptomatic subjects approaching adulthood. Further studies are needed in symptomatic children within this age group.

A paracrine requirement for hedgehog signalling in cancer.

Ligand-dependent activation of the hedgehog (Hh) signalling pathway has been associated with tumorigenesis in a number of human tissues. Here we show that, although previous reports have described a cell-autonomous role for Hh signalling in these tumours, Hh ligands fail to activate signalling in tumour epithelial cells. In contrast, our data support ligand-dependent activation of the Hh pathway in the stromal microenvironment. Specific inhibition of Hh signalling using small molecule inhibitors, a neutralizing anti-Hh antibody or genetic deletion of smoothened (Smo) in the mouse stroma results in growth inhibition in xenograft tumour models. Taken together, these studies demonstrate a paracrine requirement for Hh ligand signalling in the tumorigenesis of Hh-expressing cancers and have important implications for the development of Hh pathway antagonists in cancer.

Cortical processing of respiratory occlusion stimuli in children with central hypoventilation syndrome.

RATIONALE: The ability of patients with central hypoventilation syndrome (CHS) to produce and process mechanoreceptor signals is unknown. OBJECTIVES: Children with CHS hypoventilate during sleep, although they generally breathe adequately during wakefulness. Previous studies suggest that they have compromised central integration of afferent stimuli, rather than abnormal sensors or receptors. Cortical integration of afferent mechanical stimuli caused by respiratory loading or upper airway occlusion can be tested by measuring respiratory-related evoked potentials (RREPs). We hypothesized that patients with CHS would have blunted RREP during both wakefulness and sleep. METHODS: RREPs were produced with multiple upper airway occlusions and were obtained during wakefulness, stage 2, slow-wave, and REM sleep. Ten patients with CHS and 20 control subjects participated in the study, which took place at the Children's Hospital of Philadelphia. Each patient was age- and sex-matched to two control subjects. Wakefulness data were collected from 9 patients and 18 control subjects. MEASUREMENTS AND MAIN RESULTS: During wakefulness, patients demonstrated reduced Nf and P300 responses compared with control subjects. During non-REM sleep, patients demonstrated a reduced N350 response. In REM sleep, patients had a later P2 response. CONCLUSIONS: CHS patients are able to produce cortical responses to mechanical load stimulation during both wakefulness and sleep; however, central integration of the afferent signal is disrupted during wakefulness, and responses during non-REM are damped relative to control subjects. The finding of differences between patients and control subjects during REM may be due to increased intrinsic excitatory inputs to the respiratory system in this state.

Puberty and upper airway dynamics during sleep.

STUDY OBJECTIVES: The upper airway compensatory response to subatmospheric pressure loading declines with age. The epidemiology of obstructive sleep apnea suggests that sex hormones play a role in modulating upper airway function. Sex hormones increase gradually during puberty, from minimally detectable to adult levels. We hypothesized that the upper airway response to subatmospheric pressure loading decreased with increasing pubertal Tanner stage in males but remained stable during puberty in females. DESIGN: Upper airway dynamic function during sleep was measured over the course of puberty. PARTICIPANTS: Normal subjects of Tanner stages 1 to 5. MEASUREMENTS: During sleep, maximal inspiratory airflow was measured while varying the level of nasal pressure. The slope of the upstream pressure-flow relationship (SPF) was measured. RESULTS: The SPF correlated with age and Tanner stage. However, the relationship with Tanner stage became nonsignificant when the correlation due to the mutual association with age was removed. Females had a lower SPF than males. CONCLUSIONS: In both sexes, the upper airway compensatory response to subatmospheric pressure loading decreased with age rather than degree of pubertal development. Thus, changes in sex hormones are unlikely to be a primary modulator of upper airway function during the transition from childhood to adulthood. Although further studies of upper airway structural changes during puberty are needed, we speculate that the changes in upper airway function with age are due to the depressant effect of age on ventilatory drive, leading to a decrease in upper airway neuromotor tone.

Effect of sleep stage on breathing in children with central hypoventilation.

The early literature suggests that hypoventilation in infants with congenital central hypoventilation syndrome (CHS) is less severe during rapid eye movement (REM) than during non-REM (NREM) sleep. However, this supposition has not been rigorously tested, and subjects older than infancy have not been studied. Given the differences in anatomy, physiology, and REM sleep distribution between infants and older children, and the reduced number of limb movements during REM sleep, we hypothesized that older subjects with CHS would have more severe hypoventilation during REM than NREM sleep. Nine subjects with CHS, aged (mean +/- SD) 13 +/- 7 yr, were studied. Spontaneous ventilation was evaluated by briefly disconnecting the ventilator under controlled circumstances. Arousal was common, occurring in 46% of REM vs. 38% of NREM trials [not significant (NS)]. Central apnea occurred during 31% of REM and 54% of NREM trials (NS). Although minute ventilation declined precipitously during both REM and NREM trials, hypoventilation was less severe during REM (drop in minute ventilation of 65 +/- 23%) than NREM (drop of 87 +/- 16%, P = 0.036). Despite large changes in gas exchange during trials, there was no significant change in heart rate during either REM or NREM sleep. We conclude that older patients with CHS frequently have arousal and central apnea, in addition to hypoventilation, when breathing spontaneously during sleep. The hypoventilation in CHS is more severe during NREM than REM sleep. We speculate that this may be due to increased excitatory inputs to the respiratory system during REM sleep.

Cortical processing of respiratory afferent stimuli during sleep in children with the obstructive sleep apnea syndrome.

STUDY OBJECTIVES: Children with the obstructive sleep apnea syndrome (OSAS) have blunted upper airway responses to negative pressure, but the underlying cause remains unknown. Cortical processing of respiratory afferent information can be tested by measuring respiratory-related evoked potentials (RREPs). We hypothesized that children with OSAS have blunted RREP responses compared to normal children during sleep. DESIGN: During sleep, RREPs were obtained from EEG electrodes Fz, Cz, Pz during stage 2 sleep, slow wave sleep (SWS), and REM sleep. RREPs were produced with multiple short occlusions of the upper airway. SETTING: Sleep laboratory. PARTICIPANTS: 9 children with OSAS and 12 normal controls. MEASUREMENTS AND RESULTS: Children with OSAS had significantly decreased evoked K-complex production in stage 2 sleep and slow wave sleep and significantly reduced RREP N350 and P900 components in slow wave sleep. There were no significant differences in any of the measured RREP components in stage 2 sleep, and the only REM difference was decreased P2 amplitude. CONCLUSIONS: Results indicate that in children with OSAS, cortical processing of respiratory-related information measured with RREPs persists throughout sleep; however, RREPs during SWS are blunted compared to those seen in control children. Possible causes for this difference include a congenital deficit in neural processing reflective of a predisposition to develop OSAS, or changes in the upper airway rendering the airway less capable of transducing pressure changes following occlusion. Further research is required to evaluate RREPs after effective surgical treatment of OSAS in children, in order to distinguish between these alternatives.