A CRISPR Path to Finding Vulnerabilities and Solving Drug Resistance: Targeting the Diverse Cancer Landscape and Its Ecosystem.

Cancer is the second leading cause of death globally, with therapeutic resistance being a major cause of treatment failure in the clinic. The dynamic signaling that occurs between tumor cells and the diverse cells of the surrounding tumor microenvironment actively promotes disease progression and therapeutic resistance. Improving the understanding of how tumors evolve following therapy and the molecular mechanisms underpinning de novo or acquired resistance is thus critical for the identification of new targets and for the subsequent development of more effective combination regimens. Simultaneously targeting multiple hallmark capabilities of cancer to circumvent adaptive or evasive resistance may lead to significantly improved treatment response in the clinic. Here, the latest applications of functional genomics tools, such as clustered regularly interspaced short palindromic repeats (CRISPR) editing, to characterize the dynamic cancer resistance mechanisms, from improving the understanding of resistance to classical chemotherapeutics, to deciphering unique mechanisms that regulate tumor responses to new targeted agents and immunotherapies, are discussed. Potential avenues of future research in combating therapeutic resistance, the contribution of tumor-stroma signaling in this setting, and how advanced functional genomics tools can help streamline the identification of key molecular determinants of drug response are explored.

Cancer-Associated Fibroblasts in Pancreatic Ductal Adenocarcinoma Determine Response to SLC7A11 Inhibition.

Cancer-associated fibroblasts (CAF) are major contributors to pancreatic ductal adenocarcinoma (PDAC) progression through protumor signaling and the generation of fibrosis, the latter of which creates a physical barrier to drugs. CAF inhibition is thus an ideal component of any therapeutic approach for PDAC. SLC7A11 is a cystine transporter that has been identified as a potential therapeutic target in PDAC cells. However, no prior study has evaluated the role of SLC7A11 in PDAC tumor stroma and its prognostic significance. Here we show that high expression of SLC7A11 in human PDAC tumor stroma, but not tumor cells, is independently prognostic of poorer overall survival. Orthogonal approaches showed that PDAC-derived CAFs are highly dependent on SLC7A11 for cystine uptake and glutathione synthesis and that SLC7A11 inhibition significantly decreases CAF proliferation, reduces their resistance to oxidative stress, and inhibits their ability to remodel collagen and support PDAC cell growth. Importantly, specific ablation of SLC7A11 from the tumor compartment of transgenic mouse PDAC tumors did not affect tumor growth, suggesting the stroma can substantially influence PDAC tumor response to SLC7A11 inhibition. In a mouse orthotopic PDAC model utilizing human PDAC cells and CAFs, stable knockdown of SLC7A11 was required in both cell types to reduce tumor growth, metastatic spread, and intratumoral fibrosis, demonstrating the importance of targeting SLC7A11 in both compartments. Finally, treatment with a nanoparticle gene-silencing drug against SLC7A11, developed by our laboratory, reduced PDAC tumor growth, incidence of metastases, CAF activation, and fibrosis in orthotopic PDAC tumors. Overall, these findings identify an important role of SLC7A11 in PDAC-derived CAFs in supporting tumor growth. SIGNIFICANCE: This study demonstrates that SLC7A11 in PDAC stromal cells is important for the tumor-promoting activity of CAFs and validates a clinically translatable nanomedicine for therapeutic SLC7A11 inhibition in PDAC.

Health care utilization by women sexual assault survivors after emergency care: Results of a multisite prospective study.

BACKGROUND: Approximately, 100,000 US women receive emergency care after sexual assault each year, but no large-scale study has examined the incidence of posttraumatic sequelae, receipt of health care, and frequency of assault disclosure to providers. The current study evaluated health outcomes and service utilization among women in the 6 weeks after sexual assault. METHODS: Women ≥18 years of age presenting for emergency care after sexual assault to twelve sites were approached. Among those willing to be contacted for the study (n = 1080), 706 were enrolled. Health outcomes, health care utilization, and assault disclosure were assessed via 6 week survey. RESULTS: Three quarters (76%) of women had posttraumatic stress, depression, or anxiety, and 65% had pain. Less than two in five reported seeing health care provider; receipt of care was not related to substantive differences in symptoms and was less likely among Hispanic women and women with a high school education or less. Nearly one in four who saw a primary care provider did not disclose their assault, often due to shame, embarrassment, or fear of being judged. CONCLUSION: Most women receiving emergency care after sexual assault experience substantial posttraumatic sequelae, but health care in the 6 weeks after assault is uncommon, unrelated to substantive differences in need, and limited in socially disadvantaged groups. Lack of disclosure to primary care providers was common among women who did receive care.

Jumping to conclusions in the less-delusion-prone? Further evidence from a more reliable beads task.

BACKGROUND: A single meta-analysis has found that healthy people with higher delusion-proneness tend to gather less information (i.e., make fewer draws to decision, or DTD) on the beads task, although the findings of contributing studies were mixed, and the pooled effect size was small. However, using a new and more reliable "distractor sequences" beads task, we recently found a positive relationship between delusion-proneness and DTD in a healthy sample. In the current study, we re-tested this relationship in a new sample, and tested the possibility that the relationship is driven by participant's ability to understand and use odds or likelihood information ("odds literacy"). METHODS: Healthy participants (N = 167) completed the distractor sequences beads task, the Peters Delusions Inventory (PDI) which measures delusion-proneness, a measure of odds literacy, and the Depression, Anxiety, and Stress scale. RESULTS: PDI and DTD were positively correlated, and comparing PDI quartiles on DTD confirmed a statistically significant trend of increasing DTD with PDI quartile. Odds literacy was positively rather than negatively associated with both DTD and PDI. Anxiety was positively correlated with PDI and DTD. CONCLUSIONS: We replicated our earlier finding that DTD and delusion-proneness were positively related in a non-clinical sample, but found that increased odds-literacy did not drive lower PDI and DTD, and hence did not explain their covariance. It is possible however that anxiety and co-occurring risk aversion drive increased delusion-proneness and information-gathering, potentially accounting for the positive relationship between PDI and DTD.

Jumping to conclusions in the less-delusion-prone? Preliminary evidence from a more reliable beads task.

BACKGROUND AND OBJECTIVES: Several meta-analyses have shown that people with psychosis tend to gather less information (i.e., they make fewer draws to decision, or DTD) on the beads task than healthy controls. A single meta-analysis has also found a small negative association between delusion-proneness and DTD in healthy samples, but with considerable heterogeneity. METHODS: We used the new and more reliable "distractor sequences" beads task to clarify the nature of the relationship between delusion-proneness and DTD in a healthy sample. Healthy participants (N = 203) completed the distractor sequences beads task and the Peters Delusions Inventory (PDI), which measures delusion-proneness. RESULTS: PDI and DTD were positively correlated, and those who jumped to conclusions (DTD ≤ 2) had lower PDI than those who did not. Comparing PDI quartiles on DTD provided some evidence the positive association did not extend to the highest PDI quartile. We found that DTD and delusion-proneness were positively related in our non-clinical sample, which was unexpected. LIMITATIONS: Results need replication with a clinical sample. CONCLUSIONS: Considering the well-established association between the JTC bias and clinical delusions, the current finding may reflect a relationship that differs between non-clinical and clinically significant delusional groups, or one which reverses sign at some level of delusion-proneness.

Towards a reliable repeated-measures beads task for assessing the jumping to conclusions bias.

The jumping to conclusions bias (JTC), in which some people gather less information than others before making a decision, has been linked to delusions in psychosis. JTC is usually identified via the beads task, in which a sequence of beads (the "target" sequence) is used to measure the amount of evidence participants require before making a decision. Yet, despite its common use, the reliability of the task has never been properly investigated. We investigated its reliability, and tested an alternate version which used distractor sequences to obfuscate the target sequence. Healthy participants (N = 212) were randomised into two groups. One group completed ten trials using the target sequence, while the other completed ten trials of the target sequence and three distractor sequences. Our data indicated the standard task may not be reliable over repeated measures, but that by including distractor sequences, the task becomes more believable, repeatable, and reliable. Additionally, excluding first-trial data (a "silent" practice trial) also improves repeatability. These improvements to the task are relevant to single trial studies, and will be especially useful to repeated-measures longitudinal, experimental, and treatment studies. Such repeated-measures studies are important for investigating the causal link between JTC and delusions.

Morningness/eveningness and the synchrony effect for spatial attention.

There is evidence that a decrease in alertness is associated with a rightward shift of attention. Alertness fluctuates throughout the day and peak times differ between individuals. Some individuals feel most alert in the morning; others in the evening. Our aim was to investigate the influence of morningness/eveningness and time of testing on spatial attention. It was predicted that attention would shift rightwards when individuals were tested at their non-optimal time as compared to tests at peak times. A crowdsourcing internet marketplace, Amazon Mechanical Turk (AMT) was used to collect data. Given questions surrounding the quality of data drawn from such virtual environments, this study also investigated the sensitivity of data to demonstrate known effects from the literature. Five-hundred and thirty right-handed participants took part between 6 am and 11 pm. Participants answered demographic questions, completed a question from the Horne and Östberg Morningness/Eveningness Scale, and performed a spatial attentional task (landmark task). For the landmark task, participants indicated whether the left or right segment of each of 72 pre-bisected lines was longer (longer side counterbalanced). Response bias was calculated by subtracting the 'number of left responses' from the 'number of right responses', and dividing by the number of trials. Negative values indicate a leftward attentional bias, and positive values a rightward bias. Well-supported relationships between variables were reflected in the dataset. Controlling for age, there was a significant interaction between morningness/eveningness and time of testing (morning=6 am-2.30 pm, evening=2.30 pm-11 pm) (p<0.05) such that there was a relative rightward shift of attention from peak to off-peak times of testing for those identifying as morning types, but not evening types. Findings support the utility of crowdsourcing internet marketplaces as data collection vehicles for research. Results also suggest that the deployment of spatial attention is modulated by an individual's peak time (morningness/eveningness) and time of testing.

Do depressive symptoms predict paranoia or vice versa?

BACKGROUND AND OBJECTIVES: Affective versus nonaffective psychoses are today no longer regarded as mutually exclusive disorders. Theorists have recently highlighted the role of affective symptoms in the formation of paranoid beliefs, particularly negative beliefs about the self, interpersonal sensitivity, sleep disturbances, and worrying, which exist along a continuum in the general population. For the present study, we tested the bidirectional causal relationships between paranoia and affect. METHOD: A large population sample (N = 2,357) was examined at three time-points (baseline, six months, two years) as to the severity of subclinical paranoid beliefs (Paranoia Checklist, PCL) and depressive symptoms (Patient Health Questionnaire-9, PHQ-9). Worrying and avoidance were measured with items from the Maladaptive and Adaptive Coping Style Questionnaire (MAX). RESULTS: Depression and paranoid symptoms were strongly cross-sectionally related (r = 0.69) and showed high stability (r > 0.72). Depressive symptoms at T2 predicted paranoid symptoms at T3 (beta = 0.16; no significant relationship from T1 to T2), whereas paranoid symptoms predicted depressive symptoms from T1 to T2 (beta = 0.09; no significant relationship from T2 to T3). LIMITATIONS: Results should be replicated in a sample of paranoid patients, as risk factors for subclinical versus manifest paranoia may differ. Some constructs were measured with single items derived from a new scale. CONCLUSIONS: The predictive association of depression to subsequent paranoia was small and confined to the long interval from T2 to T3. Treatments should target both paranoia and depression - irrespective of their causal relationship - particularly as patients with psychosis consider treatment of their emotional problems a priority.

Association of the Jumping to Conclusions and Evidence Integration Biases With Delusions in Psychosis: A Detailed Meta-analysis.

We completed a meta-analysis to investigate the relationship between delusions in psychosis and 4 cognitive biases: "jumping to conclusions" (JTC), the "bias against disconfirmatory evidence" (BADE), the "bias against confirmatory evidence" (BACE), and "liberal acceptance" (LA). Building on recent meta-analyses we compared more narrowly defined groups. We identified 35 JTC, 8 BADE, 7 BACE, and 6 LA studies for inclusion. Groups with schizophrenia who were currently experiencing delusions demonstrated greater JTC, BADE, BACE, and LA than groups with schizophrenia who were not currently experiencing delusions, who in turn demonstrated no more JTC than healthy control groups. Hence JTC, BADE, BACE, and LA co-vary with delusions in cross-sectional samples of people with schizophrenia. Groups who were experiencing delusions due to other psychiatric illnesses also demonstrated greater JTC than healthy controls, and equivalent JTC to groups with schizophrenia currently experiencing delusions. Hence JTC is associated with delusions across a range of diagnoses. Groups with other, non-delusional psychiatric illnesses demonstrated less JTC, BADE, BACE, and LA than groups with schizophrenia currently experiencing delusions, less JTC than groups experiencing delusions due to other diagnoses, and no more JTC, BADE, BACE, or LA than healthy control groups. Hence JTC, BADE, BACE, and LA were not associated with psychiatric illnesses in general. Our results indicate all 4 biases are associated with delusions specifically rather than merely with a diagnosis of schizophrenia or with being psychiatrically ill, consistent with the possibility that they contribute to delusional severity.