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Supplementary Figures - PDF file 1385K, File contains Supplementary figures of resistant cell lines, proliferation assay of a panel of cell lines, TdF binding assay transcriptional out and kinome selectivity

Supplementary Table 1 from A Fully Human Insulin-Like Growth Factor-I Receptor Antibody SCH 717454 (Robatumumab) Has Antitumor Activity as a Single Agent and in Combination with Cytotoxics in Pediatric Tumor Xenografts

Supplementary Figure Legends 1-3, Table Legend 1 from A Fully Human Insulin-Like Growth Factor-I Receptor Antibody SCH 717454 (Robatumumab) Has Antitumor Activity as a Single Agent and in Combination with Cytotoxics in Pediatric Tumor Xenografts

Supplementary Figure 1 from A Fully Human Insulin-Like Growth Factor-I Receptor Antibody SCH 717454 (Robatumumab) Has Antitumor Activity as a Single Agent and in Combination with Cytotoxics in Pediatric Tumor Xenografts

The insulin-like growth factor-I receptor (IGF-IR) and its ligands (IGF-I and IGF-II) have been implicated in the growth, survival, and metastasis of a broad range of malignancies including pediatric tumors. Blocking the IGF-IR action is a potential cancer treatment. A fully human neutralizing monoclonal antibody, SCH 717454 (19D12, robatumumab), specific to IGF-IR, has shown potent antitumor effects in ovarian cancer in vitro and in vivo. In this study, SCH 717454 was evaluated in several pediatric solid tumors including neuroblastoma, osteosarcoma, and rhabdomyosarcoma. SCH 717454 is shown here to downregulate IGF-IR as well as inhibit IGF-IR and insulin receptor substrate-1 phosphorylation in pediatric tumor cells. IGF-IR and insulin receptor substrate-1 phosphorylation in the tumor cells. In vivo, SCH 717454 exhibits activity as a single agent and significantly inhibited growth of neuroblastoma, osteosarcoma, and rhabdomyosarcoma tumor xenografts. Combination of SCH 717454 with cisplatin or cyclophosphamide enhanced both the degree and the duration of the in vivo antitumor activity compared with single-agent treatments. Furthermore, SCH 717454 treatment markedly reduced Ki-67 expression and blood vessel formation in tumor xenografts, showing that the in vivo activity is derived from its inhibition of tumor cell proliferation and angiogenesis activity. Mol Cancer Ther; 9(2); 410–8

Supplementary Figure 3 from A Fully Human Insulin-Like Growth Factor-I Receptor Antibody SCH 717454 (Robatumumab) Has Antitumor Activity as a Single Agent and in Combination with Cytotoxics in Pediatric Tumor Xenografts

Supplementary Figure 2 from A Fully Human Insulin-Like Growth Factor-I Receptor Antibody SCH 717454 (Robatumumab) Has Antitumor Activity as a Single Agent and in Combination with Cytotoxics in Pediatric Tumor Xenografts

Arterial diastolic blood pressure (DBP) greater than 25 mm Hg in infants and greater than 30 mm Hg in children greater than 1 year old during cardiopulmonary resuscitation (CPR) was associated with survival to hospital discharge in one prospective study. We sought to validate these potential hemodynamic targets in a larger multicenter cohort.Prospective observational study.Eighteen PICUs in the ICU-RESUScitation prospective trial from October 2016 to March 2020.Children less than or equal to 18 years old with CPR greater than 30 seconds and invasive blood pressure (BP) monitoring during CPR.None.Invasive BP waveform data and Utstein-style CPR data were collected, including prearrest patient characteristics, intra-arrest interventions, and outcomes. Primary outcome was survival to hospital discharge, and secondary outcomes were return of spontaneous circulation (ROSC) and survival to hospital discharge with favorable neurologic outcome. Multivariable Poisson regression models with robust error estimates evaluated the association of DBP greater than 25 mm Hg in infants and greater than 30 mm Hg in older children with these outcomes. Among 1,129 children with inhospital cardiac arrests, 413 had evaluable DBP data. Overall, 85.5% of the patients attained thresholds of mean DBP greater than or equal to 25 mm Hg in infants and greater than or equal to 30 mm Hg in older children. Initial return of circulation occurred in 91.5% and 25% by placement on extracorporeal membrane oxygenator. Survival to hospital discharge occurred in 58.6%, and survival with favorable neurologic outcome in 55.4% (i.e. 94.6% of survivors had favorable neurologic outcomes). Mean DBP greater than 25 mm Hg for infants and greater than 30 mm Hg for older children was significantly associated with survival to discharge (adjusted relative risk [aRR], 1.32; 1.01-1.74; p = 0.03) and ROSC (aRR, 1.49; 1.12-1.97; p = 0.002) but did not reach significance for survival to hospital discharge with favorable neurologic outcome (aRR, 1.30; 0.98-1.72; p = 0.051).These validation data demonstrate that achieving mean DBP during CPR greater than 25 mm Hg for infants and greater than 30 mm Hg for older children is associated with higher rates of survival to hospital discharge, providing potential targets for DBP during CPR.

ObjectivesThe COVID-19 pandemic resulted in adaptations to pediatric resuscitation systems of care. The objective of this study was to determine the temporal association between the pandemic and pediatric in-hospital cardiac arrest (IHCA) process of care metrics, cardiopulmonary resuscitation (cardiopulmonary resuscitation) quality, and patient outcomes.DesignMulticenter retrospective analysis of a dataset comprising observations of IHCA outcomes pre pandemic (March 1, 2019 to February 29, 2020) versus pandemic (March 1, 2020 to February 28, 2021).SettingData source was the ICU-RESUScitation Project ("ICU-RESUS;" NCT028374497), a prospective, multicenter, cluster randomized interventional trial.PatientsChildren (≤ 18 yr) who received cardiopulmonary resuscitation while admitted to the ICU and were enrolled in ICU-RESUS.InterventionsNone.Measurements and main resultsAmong 429 IHCAs meeting inclusion criteria, occurrence during the pandemic period was associated with higher frequency of hypotension as the immediate cause of arrest. Cardiac arrest physiology, cardiopulmonary resuscitation quality metrics, and postarrest physiologic and quality of care metrics were similar between the two periods. Survival with favorable neurologic outcome (Pediatric Cerebral Performance Category score 1-3 or unchanged from baseline) occurred in 102 of 195 subjects (52%) during the pandemic compared with 140 of 234 (60%) pre pandemic ( p = 0.12). Among survivors, occurrence of IHCA during the pandemic period was associated with a greater increase in Functional Status Scale (FSS) (i.e., worsening) from baseline (1 [0-3] vs 0 [0-2]; p = 0.01). After adjustment for confounders, IHCA survival during the pandemic period was associated with a greater increase in FSS from baseline (+1.19 [95% CI, 0.35-2.04] FSS points; p = 0.006) and higher odds of a new FSS-defined morbidity (adjusted odds ratio, 1.88 [95% CI, 1.03-3.46]; p = 0.04).ConclusionsUsing the ICU-RESUS dataset, we found that relative to the year prior, pediatric IHCA during the first year of the COVID-19 pandemic was associated with greater worsening of functional status and higher odds of new functional morbidity among survivors.

A large body of research has found that people judge bad foreseen side effects to be more intentional than good ones. While the standard interpretation of this Side-Effect Effect (SEE) takes it to show that the ordinary concept of intentionality is influenced by normative considerations, a competing account holds that it is the result of pragmatic pressure to express moral censure and, thus, that the SEE is an experimental artifact. Attempts to confirm this account have previously been unsuccessful, but Lindauer and Southwood (Am Philos Q 58(2):181–186, 2021) present a study that appears to provide support for it, by cancelling the SEE. We are not convinced. Here, we detail three studies testing their interpretation. The results indicate that it is the purported cancellation, rather than the SEE, that is an experimental artifact.

Reliable data indicates increasing numbers of patients are harmed when receiving healthcare. The landmark paper “To Err is Human” posited that adverse events occur in the context of the complexity of systems within medicine and are not due to intentional harm. Crisis resource management (CRM) originated in aviation in 1980. CRM is defined as the cognitive, social, and personal resource skills that complement technical skills and contribute to safe task performance. It is a risk-reducing strategy utilized in aviation that has led to significant reduction in human error–related airline fatalities. CRM was adopted from commercial aviation by medicine in 1990 in an attempt to improve patient safety and reduce morbidity and mortality attributed to medical errors. In the last 40 years, commercial and military aviation has standardized CRM training which has led to a track record of success in improving safety for flight crews and passengers with flight-related mortality significantly decreasing. In stark contrast, nearly 30 years of CRM training in healthcare has been highly variable in content, quality, and outcomes. In this time, the number of patient deaths attributed to medical mistakes has increased from an estimated 50,000–100,000 in 1999 to over 250,000 per year from a 2016 estimate. It is time to reassess how CRM is being deployed in healthcare. Full CRM integration will require significant cultural and embedded organizational changes. Proficient CRM skills are a necessary, but not sufficient, condition for adverse event rate reduction: CRM should not be just what we do—it has to be a part of who we are as medical professionals.

AIM: The mathematical method used to calculate chest compression (CC) rate during cardiopulmonary resuscitation varies in the literature and across device manufacturers. The objective of this study was to determine the variability in calculated CC rates by applying four published methods to the same dataset. METHODS: This study was a secondary investigation of the first 200 pediatric cardiac arrest events with invasive arterial line waveform data in the ICU-RESUScitation Project (NCT02837497). Instantaneous CC rates were calculated during periods of uninterrupted CCs. The defined minimum interruption length affects rate calculation (e.g., if an interruption is defined as a break in CCs ≥ 2 seconds, the lowest possible calculated rate is 30 CCs/min). Average rates were calculated by four methods: 1) rate with an interruption defined as ≥ 1 second; 2) interruption ≥ 2 seconds; 3) interruption ≥ 3 seconds; 4) method #3 excluding top and bottom quartiles of calculated rates. American Heart Association Guideline-compliant rate was defined as 100–120 CCs/min. A clinically important change was defined as ± 5 CCs/min. The percentage of events and epochs (30 second periods) that changed Guideline-compliant status was calculated. RESULTS: Across calculation methods, mean CC rates (118.7 – 119.5/min) were similar. Comparing all methods, 14 events (7%) and 114 epochs (6%) changed Guideline-compliant status. CONCLUSION: Using four published methods for calculating CC rate, average rates were similar, but 7% of events changed Guideline-compliant status. These data suggest that a uniform calculation method (interruption ≥ 1 second) should be adopted to decrease variability in resuscitation science.

Introduction: Early recognition of clinical decompensation and rapid intervention are essential to prevention of cardiac arrest (CA). Simulation provides a safe, realistic environment to improve performance of low frequency, high risk skills and practical knowledge. Codes on the Go (COTG) uses mobile bedside simulation emulating the most frequent causes of CA to improve early recognition of clinical changes that may lead to CA and improve performance of crucial tasks. Methods: COTG scenarios are based on our cardiac intensive care unit CA data including trends in etiology and knowledge gaps identified in code debriefs. Each simulation lasts 10 minutes (pre-brief, clinical scenario, and educational debrief). For each simulation, completion and timing of pre-identified objectives were recorded and participant feedback was obtained. Results: We conducted 56 simulations using our pilot scenario “Agitation & Bradycardia”. Of these, 11 involved staff participating for the second time. When comparing the 45 first-time participants to the 11 repeat participants, the tasks showing the largest increase in completion were identifying that dexmedetomidine (dex) can worsen bradycardia, stopping the dex infusion, and using the appropriate CPR ratio. The average time-to-task decreased for 5 of 6 tasks measured between the first-time participants and repeat participants (Figure 1). The vast majority (98%) of participants agreed that feedback was done in a constructive, non-punitive way and the simulation had no negative impact on patient care or staff workflow. Conclusions: COTG is a customizable and low-cost way to improve performance of critical skills and knowledge. Repeat participation in the same scenario improved both completion of objectives and time-to-task without impeding workflow or patient care. This pilot study demonstrated successful proof-of-concept and further implementation of arrest scenario simulations will be implemented to expand its benefit.

Arthropod-borne viruses (arboviruses) pose a significant global health burden. Among the most medically important arboviruses, dengue, Zika, Yellow fever, and Chikungunya viruses are transmitted by two mosquito species, Aedes aegypti and Aedes albopictus, with Ae. aegypti being the primary vector. In the absence of safe and effective vaccines, efforts to limit disease have focused on vector-control, typically relying on the use of insecticides. Due to environmental concerns and the increasing problem of insecticide-resistance, alternative approaches to vector-control are being explored. One promising strategy involves the release of mosquitoes infected with the bacterial endosymbiont, Wolbachia pipientis. When transinfected into Ae. aegypti, certain strains of Wolbachia have the ability to rapidly invade populations and inhibit viral transmission or reduce population density. Field trials using the Drosophila-derived wMel strain have yielded positive results. However, recent evidence suggests that the Aedes albopictus-derived wAlbB strain may be more suitable due to its broader range of thermal tolerance compared to wMel. The usefulness of Wolbachia for arbovirus suppression is the result of a complex set of molecular interactions between host, symbiont, and pathogen. Some recent research has begun to unravel certain aspects of the relationship, including the elegant way in which Wolbachia manipulates host reproduction to facilitate its invasion into naive populations. However, much about the biology of Wolbachia infection in mosquitoes remains unknown, including, crucially, the mechanism of viral inhibition. To add further complication, in addition to vectoring arboviruses, Aedes mosquitoes also carry insect-specific viruses (ISVs), some of which have an effect on arbovirus transmission while also being affected by Wolbachia themselves. A thorough understanding of the biological underpinnings of Wolbachia maintenance and viral inhibition is required in order to optimize vector-control strategies and predict the future success of field applications. microRNAs (miRNAs) are small non-coding RNA molecules involved in regulating a range of biological processes in eukaryotes and viruses. Some miRNAs have been implicated in mediating host-microbe interactions, including between Wolbachia and mosquitoes. Using a wAlbB-transinfected Ae. aegypti mosquito line, we investigated the effect of wAlbB on the host miRNA expression profile, and used miRNA inhibitors of selected differentially expressed miRNAs to explore their effect on host longevity, innate immune function, and wAlbB maintenance. We also created a stably transinfected Ae. aegypti Aag2.wAlbB cell line using wAlbB from Ae. albopictus Aa23 cells to investigate the effect of transinfection on wAlbB gene expression. We observed a general downregulation of gene expression in wAlbB following transinfection, and identified several differentially expressed genes relating to membrane integrity, replication and biosynthesis, as well as several genes associated with prophage WOAlbB. Furthermore, we characterised a novel insect-specific positive-sense RNA negev-like virus, present in Aa23 cells and showed that it is inhibited by wAlbB in Aag2 cells but not in Aa23 cells. The results presented here serve to advance our current understanding of the intimate symbiotic relationship between wAlbB and Ae. aegypti.

TRK kinases (TRK) are clinically validated targets for cancers harboring NTRK gene fusions. However, approved TRK inhibitors (TRKi) give rise to mutations reducing long-term clinical efficacy, and display limited CNS penetration that may impact their ability to address primary brain tumors (PBT), including high-grade gliomas. Second generation compounds in development address some resistance mutations, but are not sufficiently CNS penetrant. PBI-200 is a novel, orally active TRK inhibitor designed to overcome resistance, with high CNS penetration. In vitro assays demonstrate picomolar- to nanomolar potency against both wild-type TRK and the major acquired resistance mutations. A BaF3 xenograft model encoding the LMNA-NTRK1 gene fusion and G595R solvent front mutation showed superior efficacy with PBI-200 relative to first-generation compounds larotrectinib and entrectinib, and equivalent efficacy to the second-generation compound selitrectinib, in terms of tumor growth inhibition. Importantly, drug concentrations in the brains of PBI-200 treated mice showed an approximately 4-fold brain/plasma ratio (BPR). In contrast, BPR of other TRKi were < 0.17, indicating that only PBI-200 achieved sustained drug levels in brain. PBI-200 demonstrated statistically superior CNS efficacy and survival in a KM12-Luc intracranial murine model, as compared to other TRKi (chi-squared 45.6, p < 0.0001). By day 41, 50% of mice in the PBI-200 group remained alive, whereas mice in the other treatment groups had died on or before day 32. Of note, no gross or histopathological CNS adverse effects were observed in a 14d CNS Field Observation Battery study. Finally, PBI-200 was the most selective TRKi in a 125 kinase panel (ThermoFisher) with only 1 off-target kinase inhibited > 60% at 1 micromolar. Together, these data suggest that PBI-200 has potential as a best-in-class, highly CNS-penetrant next generation TRKi. A Phase 1/2 clinical trial evaluating PBI-200 in NTRK fusion-positive patients, including patients with PBT, is ongoing (NCT04901806).

Introduction: The transition from residency to subspecialty fellowship in pediatric cardiology is challenging, with the daunting requirement to rapidly acquire a broad scope of knowledge and skill. In 2015, a pediatric cardiology boot camp was designed as an educational tool to help prepare trainees for this transition. Hypothesis: A national pediatric cardiology boot camp consistently improves knowledge and decreases anxiety for prospective fellows. Methods: In late spring each year (2015-2019), a 2.5-day intensive program was provided for trainees prior to beginning fellowship in July. Hands-on, simulation-based experiences were provided on topics including anatomy, auscultation, echocardiography, catheterization, cardiovascular intensive care, electrophysiology, heart failure, pulmonary hypertension, and cardiac surgery. Knowledge based exams and surveys were completed by each participant pre- and post-training. Pre- and post-training exam results were compared via paired t-tests and survey results were compared via Wilcoxon rank sum. Results: Over 5 years 144 participants (72 female, 50%) completed the course, representing 40 fellowship programs in the United States and Canada. In aggregate, significant improvement was seen in participants’ knowledge assessment (pre 45 ± 11% vs. post 71 ± 9%; p Conclusions: The Pediatric Cardiology Boot Camp provides a significant and reproducible educational benefit to participants nationwide. This intensive program simultaneously improves learners’ knowledge and alleviates anxiety as they transition to subspecialty training.

Shock is defined as an imbalance between oxygen delivery (DO2) and oxygen consumption (VO2). Oxygen demand greater than oxygen delivery leads to a severe decrease in tissue oxygenation, nutrient delivery, and eventually cell death. Shock can damage all tissues and quickly progress to multiorgan failure. This is why early identification and treatment of shock is critical to prevent irreversible cell death. This chapter provides an overview of the diagnosis and management of this complex entity.

Background High-quality cardiopulmonary resuscitation is associated with improved survival and neurological outcomes after cardiac arrest. Unfortunately, health care professionals frequently do not perform resuscitation within guidelines after life-support training. Objectives To determine if brief intermittent training in cardiopulmonary resuscitation could improve nurses’ skills to perform high-quality resuscitation 70% or more of the time during 2 minutes of cardiopulmonary resuscitation after 3 training sessions. Methods In a prospective single-center quality improvement program, pediatric critical care nurses had monthly training in cardiopulmonary resuscitation. A portable manikin/defibrillator with a chest compression sensor was used to provide corrective audiovisual feedback to optimize resuscitation skills. Resuscitation was practiced on an adult manikin. Target goals were compression depth 2 in or greater at a compression rate of 100/min to 120/minute. Percentage of time in the target range and mean compression depth and rate were recorded. Data were collected every other month. The percentage of time both compression rate and depth were in the target range was compared among nurses with different total numbers of training sessions. Results Of the 62 nurses who participated in the training, 48 had data collected. The median percentage of time in the target range improved from 29% with no training to 46% after 1 session, 54% after 2 sessions, 68% after 3 sessions, and 74% after 4 sessions (P = .001). Compression depth increased with the number of training sessions (P = .002). Conclusions This training program in cardiopulmonary resuscitation yielded significant skill improvement and retention.

A new subseries of substituted piperidines as p53-HDM2 inhibitors exemplified by 21 has been developed from the initial lead 1. Research focused on optimization of a crucial HDM2 Trp23-ligand interaction led to the identification of 2-(trifluoromethyl)thiophene as the preferred moiety. Further investigation of the Leu26 pocket resulted in potent, novel substituted piperidine inhibitors of the HDM2-p53 interaction that demonstrated tumor regression in several human cancer xenograft models in mice. The structure of HDM2 in complex with inhibitors 3, 10, and 21 is described.

Background Mastery of cardiac auscultation is a core component of cardiology training. New computer-based auscultation teaching tools have been developed to help teach this skill to trainees. OBJECTIVE: To assess incoming pediatric cardiology fellows’ auscultation skills and evaluate a phone-based application (app) to teach auscultation. Methods At the annual national Pediatric Cardiology Fellows’ Boot Camp at Stanford, the eMurmur© app was utilized to teach core auscultation skills to incoming pediatric cardiology fellows. In groups of 5 trainees each, attendees were taught basic auscultation skills in 30-minute blocks. Auscultation teaching centered around distinguishing innocent (INN) vs pathological murmurs (PATH) and focused on key diagnoses and murmurs most frequently encountered by pediatric cardiologists. Pre- and post-training assessments and surveys [Likert scoring 1-5] were administered to all attendees and were analyzed via paired t-test and sign rank test. Results A total of 40 incoming cardiology fellows from 35 training programs attended the Boot Camp at Stanford University in Palo Alto, CA from June 1-3, 2019. All trainees completed the pre- and post-training assessments. Prior to the course, trainees were less likely to correctly distinguish innocent murmurs vs. pathologic murmurs (INN 77 ± 26 % vs. PATH 92 ± 11%; p = 0.001). 73% of trainees reported the auscultation session improved their auscultation skills. Though there was no difference in overall test scores (PRE 82 ± 16% vs. POST 85 ± 17%; p = 0.13), after the training the trainees’ self-reported comfort in identifying the auscultatory findings for the most common lesions seen in pediatric cardiology were higher (PRE 3.3 ± 0.9 vs. POST 3.8 ± 0.8; p = 0.03). Comfort scores improved most for distinguishing systolic vs. diastolic murmurs (PRE 3.7 ± 0.7 vs. POST 4.1 ± 0.6 POST; p = 0.05) and gallops (PRE 3.3 ± 0.09 POST 3.8 ± 0.7; p = 0.005). Conclusions The use of an app-based teaching tool is an effective method to teach auscultation skills to incoming pediatric cardiology fellows. The app can improve fellows’ comfort with identifying the most common lesions seen in pediatric cardiology.

In 2015, it became clear to the Library and Academic Technology group at Washington College that the current asset management and support ticketing system was not adequate to handle the demands of a growing campus and catalog of devices to support. That service, GLPi, was supported in-house, and the senior systems engineer in charge of supporting the system had left the college. There was a need to move to a fully supported cloud-based service that could keep up with ever-increasing demands. Through a several month-long RFP process, four vendors were short-listed. Samanage, with its integrated asset tracking and ticketing system at an affordable price point, was the clear winner. Library and Academic Technology needed to adopt the new system at an accelerated pace due to an impending asset audit that was to be completed in the summer of 2016. Samanage's implementation team was incredibly helpful and worked with us to meet all our deadlines. Since 2016, the service has expanded to be used by multiple groups on campus to streamline common tasks and services. There are still many features and processes within Samanage we do not yet use, but have plans to implement in the future.

[Image: see text] The emergence and evolution of new immunological cancer therapies has sparked a rapidly growing interest in discovering novel pathways to treat cancer. Toward this aim, a novel series of pyrrolidine derivatives (compound 5) were identified as potent inhibitors of ERK1/2 with excellent kinase selectivity and dual mechanism of action but suffered from poor pharmacokinetics (PK). The challenge of PK was overcome by the discovery of a novel 3(S)-thiomethyl pyrrolidine analog 7. Lead optimization through focused structure–activity relationship led to the discovery of a clinical candidate MK-8353 suitable for twice daily oral dosing as a potential new cancer therapeutic.

BACKGROUND. Constitutive activation of ERK1/2 occurs in various cancers, and its reactivation is a well-described resistance mechanism to MAPK inhibitors. ERK inhibitors may overcome the limitations of MAPK inhibitor blockade. The dual mechanism inhibitor SCH772984 has shown promising preclinical activity across various BRAFV600/RAS-mutant cancer cell lines and human cancer xenografts. METHODS. We have developed an orally bioavailable ERK inhibitor, MK-8353; conducted preclinical studies to demonstrate activity, pharmacodynamic endpoints, dosing, and schedule; completed a study in healthy volunteers (P07652); and subsequently performed a phase I clinical trial in patients with advanced solid tumors (MK-8353-001). In the P07652 study, MK-8353 was administered as a single dose in 10- to 400-mg dose cohorts, whereas in the MK-8353-001 study, MK-8353 was administered in 100- to 800-mg dose cohorts orally twice daily. Safety, tolerability, pharmacokinetics, pharmacodynamics, and antitumor activity were analyzed. RESULTS. MK-8353 exhibited comparable potency with SCH772984 across various preclinical cancer models. Forty-eight patients were enrolled in the P07652 study, and twenty-six patients were enrolled in the MK-8353-001 study. Adverse events included diarrhea (44%), fatigue (40%), nausea (32%), and rash (28%). Dose-limiting toxicity was observed in the 400-mg and 800-mg dose cohorts. Sufficient exposure to MK-8353 was noted that correlated with biological activity in preclinical data. Three of fifteen patients evaluable for treatment response in the MK-8353-001 study had partial response, all with BRAFV600-mutant melanomas. CONCLUSION. MK-8353 was well tolerated up to 400 mg twice daily and exhibited antitumor activity in patients with BRAFV600-mutant melanoma. However, antitumor activity was not particularly correlated with pharmacodynamic parameters. TRIAL REGISTRATION. ClinicalTrials.gov NCT01358331. FUNDING. Merck Sharp & Dohme Corp., a subsidiary of Merck & Co. Inc., and NIH (P01 CA168585 and R35 CA197633).

In the course of 3 years, Library and Academic Technology (LAT) at Washington College (WC) managed to create successful academic learning environments that benefit both teachers and learners. Our existing classroom audio/visual (A/V) systems were old, convoluted and unreliable. Additionally, the furniture was designed for utility and cost efficiency as opposed to student creativity and collaboration.The Center for Teaching and Learning at WC funded a pilot updated classroom designed with modern, reliable A/V and student collaboration as the focus. This update included bring-your-own-device (BYOD) capabilities, updated projection equipment, and a simple-to-use control interface. We also removed the old armchairs which had small flip-up writing surfaces and replaced them with Node Chairs which have larger writing surfaces in addition to a wheeled base and a storage area for bags or books. The room redesign was an overwhelming success.Following the pilot, the new classroom design was adopted as the standard and implemented campus wide. Classroom installations were completed during breaks in the academic year, so as to not to disrupt instruction. Performing most technology installations in-house helped keep labor costs to a minimum. Furniture updates in learning spaces are moving more slowly due to budget constraints and coordination difficulties with other campus departments. Over 80% of the 53 learning spaces on campus have been updated. Five spaces have received a furniture redesign with a focus on active learning. Future updates and plans for this series of projects includes incorporating wireless presentation capabilities as well as working with the finance office to establish a replacement cycle for classroom furniture and technologies.

Fusions of NTRK 1, 2 and3 genes encoding the TRK family of receptor tyrosine kinases (TrkA, B and C, respectively) have been reported in 1-3% of all human cancers including lung and breast cancers. NTRK gene fusions are also reported in lung and breast tumors metastatic to the brain as well as in primary brain tumors including glioblastoma (3%), astrocytoma (3%) and pediatric glioma (40%). These gene fusions result in constitutive TRK kinase activity and act as oncogenic drivers of disease. First generation TRK kinase inhibitors have demonstrated clinical proof of concept in patients with tumors bearing NTRK gene fusions. PBI-200 is a novel, selective pan-TRK kinase inhibitor. In biochemical assays it inhibits TrkA, B and C with IC50 values of 0.45, 2.2 and 1.9 nM, respectively. This molecule is highly selective against a panel of 122 other protein kinases tested with the exception of Ros1 (IC50= 31 nM). Importantly, PBI-200 retains potency against resistance mutations reported in patients receiving first generation TRK kinase inhibitors. Biochemical IC50 values are 3.4 nM for the TrkA G595R gatekeeper mutation and 10 nM for the TrkA G667C mutant. PBI-200 potently inhibits proliferation of human tumor cell lines expressing TRK fusions including the KM-12 colorectal cancer cell line (TPM3-NTRK1 fusion; EC50 value = 22 nM) and the MO-91 acute myeloid leukemia cell line (ETV6-NTRK3 fusion; EC50 value = 3.5 nM). It also potently inhibits proliferation of rat BaF3 cells expressing oncogenic NTRK fusions and their resistant variants. In a subcutaneous xenograft model of KM-12 colorectal cancer, PBI-200 induces tumor stasis when dosed intraperitoneally at 15 or 30 mg/kg twice daily (93 and 100% tumor growth inhibition, respectively). The activity of PBI-200 was comparable or superior to that seen with the first generation TRK kinase inhibitors used as comparators in this model. PBI-200 has oral bioavailability in both rat and mouse. Importantly, PBI-200 demonstrates excellent brain penetration in rodent species, with brain/plasma AUC ratios of 3.9 in the mouse and 3.2 in the rat. This is in contrast to the poor brain penetration observed with the first generation TRK kinase inhibitors, larotrectinib and entrectinib.PBI-200 is efficacious in an intracranial KM-12 xenograft model when dosed intraperitoneally or orally, significantly slowing tumor growth and extending survival. Based on the activity of PBI-200 against clinically-relevant resistance mutations and its excellent brain penetration, this molecule has potential to be a next-generation TRK kinase inhibitor especially for primary brain tumors or brain metastatic lesions that harbor a NTRK gene fusion. PBI-200 is currently undergoing further characterization in IND enabling studies. Citation Format: Anthony Regina, Aram Elagoz, Vincent Albert, Jonathan Boudreault, Hong Wang, Marc Ouellet, Nicolas Brunei-Latour, Edith Bellavance, Peter White, Stephane Ciblat, W. Robert Bishop, Kollol Pal. PBI-200: A novel, brain penetrant, next generation pan-TRK kinase inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2198.

The high frequency of activating RAS or BRAF mutations in cancer provides strong rationale for targeting the mitogen-activated protein kinase (MAPK) pathway. Selective BRAF and MAP-ERK kinase (MEK) inhibitors have shown clinical efficacy in patients with melanoma. However, the majority of responses are transient, and resistance is often associated with pathway reactivation of the extracellular signal-regulated kinase (ERK) signaling pathway. Here, we describe the identification and characterization of SCH772984, a novel and selective inhibitor of ERK1/2 that displays behaviors of both type I and type II kinase inhibitors. SCH772984 has nanomolar cellular potency in tumor cells with mutations in BRAF, NRAS, or KRAS and induces tumor regressions in xenograft models at tolerated doses. Importantly, SCH772984 effectively inhibited MAPK signaling and cell proliferation in BRAF or MEK inhibitor–resistant models as well as in tumor cells resistant to concurrent treatment with BRAF and MEK inhibitors. These data support the clinical development of ERK inhibitors for tumors refractory to MAPK inhibitors. Significance: BRAF and MEK inhibitors have activity in MAPK-dependent cancers with BRAF or RAS mutations. However, resistance is associated with pathway alterations resulting in phospho-ERK reactivation. Here, we describe a novel ERK1/2 kinase inhibitor that has antitumor activity in MAPK inhibitor-naïve and MAPK inhibitor-resistant cells containing BRAF or RAS mutations. Cancer Discov; 3(7); 742–50. ©2013 AACR. See related commentary by Nissan et al., p. 719 This article is highlighted in the In This Issue feature, p. 705

Hutchinson–Gilford progeria syndrome (HGPS) is an extremely rare, fatal, segmental premature aging syndrome caused by a mutation in LMNA that produces the farnesylated aberrant lamin A protein, progerin. This multisystem disorder causes failure to thrive and accelerated atherosclerosis leading to early death. Farnesyltransferase inhibitors have ameliorated disease phenotypes in preclinical studies. Twenty-five patients with HGPS received the farnesyltransferase inhibitor lonafarnib for a minimum of 2 y. Primary outcome success was predefined as a 50% increase over pretherapy in estimated annual rate of weight gain, or change from pretherapy weight loss to statistically significant on-study weight gain. Nine patients experienced a ≥50% increase, six experienced a ≥50% decrease, and 10 remained stable with respect to rate of weight gain. Secondary outcomes included decreases in arterial pulse wave velocity and carotid artery echodensity and increases in skeletal rigidity and sensorineural hearing within patient subgroups. All patients improved in one or more of these outcomes. Results from this clinical treatment trial for children with HGPS provide preliminary evidence that lonafarnib may improve vascular stiffness, bone structure, and audiological status.

Abrogation of p53 function occurs in almost all human cancers, with more than 50% of cancers harboring inactivating mutations in p53 itself. Mutation of p53 is indicative of highly aggressive cancers and poor prognosis. The vast majority of mutations in p53 occur in its core DNA binding domain (DBD) and result in inactivation of p53 by reducing its thermodynamic stability at physiological temperature. Here, we report a small molecule, SCH529074, that binds specifically to the p53 DBD in a saturable manner with an affinity of 1-2 microm. Binding restores wild type function to many oncogenic mutant forms of p53. This small molecule reactivates mutant p53 by acting as a chaperone, in a manner similar to that previously reported for the peptide CDB3. Binding of SCH529074 to the p53 DBD is specifically displaced by an oligonucleotide with a sequence derived from the p53-response element. In addition to reactivating mutant p53, SCH529074 binding inhibits ubiquitination of p53 by HDM2. We have also developed a novel variant of p53 by changing a single amino acid in the core domain of p53 (N268R), which abolishes binding of SCH529074. This amino acid change also inhibits HDM2-mediated ubiquitination of p53. Our novel findings indicate that through its interaction with p53 DBD, SCH529074 restores DNA binding activity to mutant p53 and inhibits HDM2-mediated ubiquitination.

Exercise stimulus–response relationships for airway blood supply and dimensions have not been described in mammalian species. These relationships are vital for postulates concerning integrated reflex factors normally controlling the airways and which may underlie the asthma syndromes of exercise. This study defines airways stimulus–response relationships in exercising sheep. Ewes between 35 and 40 kg were instrumented at left thoracotomy under thiopentone/isoflurane general anaesthesia. Pulsed Doppler ultrasonic transducers were mounted on the bronchial artery, and transit-time plus single-crystal sonomicrometers on the left main bronchus. These recorded simultaneously and continuously bronchial blood flow (Q br ) and conductance (C br ), bronchial circumference (Circ br ) and wall thickness (Th br ). In Protocol 1 (P1), four sheep ran duplicate 5 min protocols on a horizontal treadmill at continuous step-up-and-down speeds of 1 min duration, namely, 0.8, 1.6, 2.2, 1.6 and 0.8 mph (moderate exercise), followed by 10 min recovery. In P2, four sheep ran duplicate 2 min protocols at constant 4 mph (strenuous exercise), and in P3, one sheep ran duplicate protocols each of 3 min at 2.2, 4.4 and 6 mph (severe exercise). Regression analysis and repeated measures ANOVA were used to assess differences between times, runs and exercise intensity. In P1, airway effects were directly related to graded exercise effort sustained over 5 min. Peak effects occurred at 2.2 mph, except for Th br . Heart rate and P a rose (to 156% and 111% of resting, respectively), and Q br and C br fell (to 83% and 75%; both P br fell to 96% ( P =0.02), and Th br rose at low speeds early and late, and thinned at the highest speed. In P2 and P3 for all variables the steady-state effects were systematically greater than for P1 (4.4 mph: C br to 43%, Circ br to 93%; 6.6 mph: C br to 25%, Circ br to 82%). There was no significant recovery hyperaemia, but there was residual post-exercise bronchoconstriction. The exercise stimulus–response relationships from rest to a maximal 6 mph for sheep airway circumference and its bronchial circulation are inverse and functionally constrictor.

Background : During exercise and recovery the transient and steady-state changes in autonomic activity regulating lower airway blood flow and dimensions are unknown. The aim of this study was to define changes in bronchial blood flow ( Q br ) and dimensions during moderate and strenuous exercise, and to analyse the role of vagal and sympathetic nerves. Methods : Nine ewes (34–44 kg) underwent left thoracotomy during general anaesthesia (thiopentone/isoflurane) and either (5 sheep=Group 1) a pulsed Doppler transducer was placed on the bronchial artery, or (4 sheep=Group 2) a pulsed Doppler transducer was placed on the bronchial artery, and transit-time and single crystal sonomicrometers were mounted on the left main bronchus. These measured continuously Q br , bronchial circumference ( Circ br ) and wall thickness ( Th br ). Aortic pressure ( P a ) and central venous pressure catheters were placed in the superficial cervical artery and vein. Trained sheep exercised on a horizontal treadmill, i.e. Group 1, moderate exercise 2.2 mph over 1.6, 6 min recovery, for analysis of changes in Q br before and after cholinoceptor blockade; Group 2, strenuous exercise 4.4 mph over 2, 10 min recovery for analysis of changes in Q br and airway dimensions, before and after cholinoceptor plus α 1 -, α 2 -adrenoceptor blockade. β -adrenoceptor systems were intact. Results : In Group 1 during moderate exercise P a and heart rate (HR) rose. Q br and blood flow conductance ( C br ) fell immediately to 83% ( P 0.001 ) before returning toward resting levels, but fell when exercise ceased to 89% ( P 0.01 ) before recovering. Prior cholinoceptor blockade abolished the immediate fall in Q br and C br , but not the recovery vasoconstriction. Later in recovery the bronchial bed dilated progressively over 6 min ( P 0.05 ) . In Group 2 during strenuous exercise P a and HR rose substantially. Q br and C br fell to 68% and 54% ( P 0.001 ) , respectively, and there was early vasoconstriction in recovery. Circ br fell immediately and remained at 93% ( P 0.01 ) , and did not recover fully when exercise ceased. Th br did not change during or after exercise. Prior cholinoceptor plus α -adrenoceptor block caused P a and Q br to fall slightly during exercise, but the bronchovascular constriction during and after exercise was abolished, as was circumferential shortening in the airway. Conclusions : At exercise onset and steady-state, resetting the arterial baroreflex upward in sheep increases parasympathetic cholinergic vasoconstrictor activity and causes bronchial wall and bronchovascular smooth muscle contraction in concert with sympathetic adrenergic constriction of systemic vascular beds. Whether the known sigmoid baroreflex control of tracheal smooth muscle tension at rest is extended to tracheobronchial smooth muscle and its circulation during exercise is yet to be determined.

Some proteins undergo posttranslational modification by the addition of an isoprenyl lipid (farnesyl- or geranylgeranyl-isoprenoid) to a cysteine residue proximal to the C terminus. Protein isoprenylation promotes membrane association and contributes to protein-protein interactions. Farnesylated proteins include small GTPases, tyrosine phosphatases, nuclear lamina, cochaperones, and centromere-associated proteins. Prenylation is required for the transforming activity of Ras. Because of the high frequency of Ras mutations in cancer, farnesyl transferase inhibitors (FTIs) were investigated as a means to antagonize Ras function. Evaluation of FTIs led to the finding that both K- and N-Ras are alternatively modified by geranylgeranyl prenyltransferase-1 in FTI-treated cells. Geranylgeranylated forms of Ras retain the ability to associate with the plasma membrane and activate substrates. Despite this, FTIs are effective at inhibiting the growth of human tumor cells in vitro, suggesting that activity is dependent on blocking the farnesylation of other proteins. FTIs also inhibit the in vivo growth of human tumor xenografts and sensitize these models to chemotherapeutics, most notably taxanes. Several FTIs have entered clinical trials for various cancer indications. In some clinical settings, primarily hematologic malignancies, FTIs have displayed evidence of single-agent activity. Clinical studies in progress are exploring the antitumor activity of FTIs as single agents and in combination. This review will summarize the basic biology of FTIs, their antitumor activity in preclinical models, and the current status of clinical studies with these agents.

Lonafarnib (SCH66336) is a farnesyl transferase inhibitor (FTI) that inhibits the post-translational lipid modification of H-Ras and other farnesylated proteins. K- and N-Ras are also substrates of farnesyl transferase; however, upon treatment with FTIs, they are alternatively prenylated by geranylgeranyl transferase-1. Despite the failure to abrogate prenylation of K- and N-Ras, growth of many tumors in preclinical models is inhibited by FTIs. This suggests that the anti-proliferative action of FTIs is dependent on blocking the farnesylation of other proteins. Rheb (Ras homologue enriched in brain) is a farnesylated small GTPase that positively regulates mTOR (mammalian target of rapamycin) signaling. We found that Rheb and Rheb2 mRNA were elevated in various tumor cell lines relative to normal cells. Peptides derived from the carboxyl termini of human Rheb and Rheb2 are in vitro substrates for farnesyl transferase but not geranylgeranyl transferase-1. Rheb prenylation in cell culture was completely inhibited by SCH66336, indicating a lack of alternative prenylation. SCH66336 treatment also inhibited the phosphorylation of S6 ribosomal protein, a downstream target of Rheb and mTOR signaling. SCH66336 did not inhibit S6 phosphorylation in cells expressing Rheb-CSVL, a mutant construct of Rheb designed to be geranylgeranylated. Importantly, expression of Rheb-CSVL also abrogated SCH66336 enhancement of tamoxifen- and docetaxel-induced apoptosis in MCF-7 breast cancer cells and ES-2 ovarian cancer cells, respectively. Further, inhibition of Rheb signaling by rapamycin treatment, small interfering RNA, or dominant negative Rheb enhanced tamoxifen- and docetaxel-induced apoptosis, similar to FTI treatment. These studies demonstrated that Rheb is modified by farnesylation, is not a substrate for alternative prenylation, and plays a role in SCH66336 enhancement of the anti-tumor response to other chemotherapeutics.

Insulin-like growth factor-I receptor (IGF-IR) plays an important role in tumor cell growth and survival. On ligand stimulation, IGF-IR, a receptor tyrosine kinase, phosphorylates tyrosine residues on two major substrates, IRS-1 and Shc, which subsequently signal through the Ras/mitogen-activated protein kinase and phosphatidylinositol 3-kinase/AKT pathways. Here, we describe the characterization of a fully human anti–IGF-IR monoclonal antibody 19D12 that inhibits IGF binding and autophosphorylation of both IGF-IR/IGF-IR homodimers and IGF-IR/insulin receptor heterodimers. 19D12 does not recognize insulin receptor homodimers. In addition to inhibiting IGF-IR autophosphorylation, 19D12 also inhibits IRS-1 phosphorylation and activation of the major downstream signaling molecules AKT and extracellular signal-regulated kinase 1/2. Furthermore, the antibody down-regulates the total IGF-IR protein level and can exhibit antibody-dependent cellular cytotoxicity activity against a non–small cell adenocarcinoma cell line in vitro in the presence of isolated human natural killer cells. 19D12 binds tightly to the receptor, with an affinity of 3.8 pmol/L as measured by KinExA. In cell culture, 19D12 inhibits proliferation and soft agar growth of various tumor cell lines. In vivo, 19D12 inhibits the tumor growth of a very aggressive human ovarian tumor xenograft model A2780. These data support the development of this anti–IGF-IR monoclonal antibody as a promising anticancer agent.

In this study, differential gene expression between normal human mammary epithelial cells and their malignant counterparts (eight well established breast cancer cell lines) was studied using Incyte GeneAlbum 1-6, which contains 65,873 cDNA clones representing 33,515 individual genes. 3,152 cDNAs showed aor =3.0-fold expression level change in at least one of the human breast cancer cell lines as compared with normal human mammary epithelial cells. Integration of breast tumor gene expression data with the genes in the tumor suppressor p53 signaling pathway yielded 128 genes whose expression is altered in breast tumor cell lines and in response to p53 expression. A hierarchical cluster analysis of the 128 genes revealed that a significant portion of genes demonstrate an opposing expression pattern, i.e. p53-activated genes are down-regulated in the breast tumor lines, whereas p53-repressed genes are up-regulated. Most of these genes are involved in cell cycle regulation and/or apoptosis, consistent with the tumor suppressor function of p53. Follow-up studies on one gene, RAI3, suggested that p53 interacts with the promoter of RAI3 and repressed its expression at the onset of apoptosis. The expression of RAI3 is elevated in most tumor cell lines expressing mutant p53, whereas RAI3 mRNA is relatively repressed in the tumor cell lines expressing wild-type p53. Furthermore, ectopic expression of RAI3 in 293 cells promotes anchorage-independent growth and small interfering RNA-mediated depletion of RAI3 in AsPc-1 pancreatic tumor cells induces cell morphological change. Taken together, these data suggest a role for RAI3 in tumor growth and demonstrate the predictive power of integrative genomics.

Modification of the ethano bridge of the core structure of the antitumor agent, SARASAR® (SCH66336) with concomitant introduction of a sulfonamide moiety off the distal piperidine afforded inhibitor 9-(S-), a compound with greatly improved PK profile. Other compounds with enhanced FPTase inhibitory activity were obtained as exemplified by amide 10-(S-) and urea 11-(S-): these compounds demonstrated activity in picomolar range.