Potential for Increasing Uptake of Radiolabeled 68Ga-DOTATOC and 123I-MIBG in Patients with Midgut Neuroendocrine Tumors Using a Histone Deacetylase Inhibitor Vorinostat.

Background: Histone deacetylase (HDAC) inhibitors have been shown in preclinical studies to upregulate norepinephrine transporters in neuroblastoma and pheochromocytoma, and somatostatin receptors in pulmonary carcinoid, small cell lung cancer, and pancreatic neuroendocrine malignancies. This pilot imaging study in humans focuses on midgut neuroendocrine carcinoma metastatic to the liver, evaluating the effect of pretreatment with the HDAC inhibitor vorinostat on uptake of 123I-MIBG and 68Ga-DOTATOC. Materials and Methods: Multiple midgut neuroendocrine liver metastases in clinically stable subjects were imaged with 123I-MIBG and 68Ga-DOTATOC before and after a 4-d course of vorinostat. Scans were performed with strict attention to detail and timed about 1 month apart occurring just before monthly long-acting octreotide administrations. Uptake changes in tumor and normal liver parenchyma were assessed on positron emission computed tomography (PET/CT) with standardized uptake values and on single photon emission computed tomography (SPECT) with qualitative ratio images. Results: The experimental units were metastatic liver lesions within patients (n = 50). There was no significant difference in administered activity or uptake time between pairs of scans for either radiotracer. Statistically significant increase in maximum standardized uptake values (SUVmax) averaged over all lesions was noted on the 68Ga-DOTATOC PET scans (+11%, p < 0.01). SUVmax in normal liver showed no significant change (p = 0.12). There was no qualitative change in uptake of 123I-MIBG after vorinostat. Conclusions: In this pilot imaging study in patients with midgut neuroendocrine liver metastases, a short course of the HDAC inhibitor vorinostat induced a statistically significant increase in SUVmax on 68Ga-DOTATOC PET/computed tomography (CT) imaging in some hepatic neuroendocrine tumor metastases. There was no significant effect of vorinostat on tumor uptake of 123I-MIBG on SPECT/CT imaging. Given the pilot nature of this trial, the findings merit further investigation with a more rigorous protocol evaluating longer pretreatment and different dosages of vorinostat or other HDAC inhibitors, as well as effects on the therapeutic capability of 177Lu- or 90Y-somatostatin analogs.

RABL6A Promotes Pancreatic Neuroendocrine Tumor Angiogenesis and Progression In Vivo.

Pancreatic neuroendocrine tumors (pNETs) are difficult-to-treat neoplasms whose incidence is rising. Greater understanding of pNET pathogenesis is needed to identify new biomarkers and targets for improved therapy. RABL6A, a novel oncogenic GTPase, is highly expressed in patient pNETs and required for pNET cell proliferation and survival in vitro. Here, we investigated the role of RABL6A in pNET progression in vivo using a well-established model of the disease. RIP-Tag2 (RT2) mice develop functional pNETs (insulinomas) due to SV40 large T-antigen expression in pancreatic islet ß cells. RABL6A loss in RT2 mice significantly delayed pancreatic tumor formation, reduced tumor angiogenesis and mitoses, and extended survival. Those effects correlated with upregulation of anti-angiogenic p19ARF and downregulation of proangiogenic c-Myc in RABL6A-deficient islets and tumors. Our findings demonstrate that RABL6A is a bona fide oncogenic driver of pNET angiogenesis and development in vivo.

Intratumoral talimogene laherparepvec injection with concurrent preoperative radiation in patients with locally advanced soft-tissue sarcoma of the trunk and extremities: phase IB/II trial.

BACKGROUND: Soft-tissue sarcomas (STS) in the extremities and trunk treated with standard-of-care preoperative external beam radiation therapy (EBRT) followed by surgical resection are associated with local and distant relapses. In preclinical studies, oncolytic virotherapy in sarcoma has demonstrated antitumor effects via direct intratumoral oncolysis and cytotoxic T-cell-mediated immune responses. Talimogene laherparepvec (TVEC) is a replication-competent, immune-enhanced, oncolytic herpes simplex virus type 1 engineered for intratumoral injection; it has been approved by the FDA for the treatment of locally advanced and metastatic melanoma. METHODS: We explored a novel combination of TVEC with standard-of-care EBRT administered preoperatively in patients with locally advanced STS of the extremities and trunk in a phase IB/II clinical trial. Thirty patients with primary STS >5 cm for which EBRT was indicated to achieve negative margins were enrolled. FDA-approved TVEC doses were used. Immune correlative studies in peripheral blood, biopsy and resected tumor tissues were performed. RESULTS: No dose-limiting toxicity was observed. Adverse events were similar to those reported in prior studies with TVEC. One patient with myxoid liposarcoma exhibited a partial response. Seven of the 29 (24%) evaluable patients achieved 95% pathological necrosis. None of the patients developed a herpes infection due to the treatment. Eight of the 29 (27%) patients developed postoperative wound complications, which is consistent with previous studies. None of the patients developed local recurrence after surgical resection of the primary sarcoma. 2-year progression-free and overall survival were 57% and 88%, respectively. Caspase-3 demonstrated increased expression of both in TVEC-treated tissue samples as compared with control samples treated with radiation alone. CONCLUSION: Preoperative intratumoral TVEC with concurrent EBRT for locally advanced STS is safe and well-tolerated. This combination treatment may enhance immune responses in some cases but did not increase the proposed rate of pathological necrosis. The Caspase-3 biomarker may be associated with a positive effect of TVEC in sarcoma tumor tissue and should be explored in future studies. TRIAL REGISTRATION NUMBER: NCT02453191.

RABL6A Is an Essential Driver of MPNSTs that Negatively Regulates the RB1 Pathway and Sensitizes Tumor Cells to CDK4/6 Inhibitors.

PURPOSE: Malignant peripheral nerve sheath tumors (MPNST) are deadly sarcomas that lack effective therapies. In most MPNSTs, the retinoblastoma (RB1) tumor suppressor is disabled by hyperactivation of cyclin-dependent kinases (CDK), commonly through loss of CDK-inhibitory proteins such as p27(Kip1). RABL6A is an inhibitor of RB1 whose role in MPNSTs is unknown. To gain insight into MPNST development and establish new treatment options, we investigated RABL6A-RB1 signaling and CDK inhibitor-based therapy in MPNSTs. EXPERIMENTAL DESIGN: We examined patient-matched MPNSTs and precursor lesions by RNA sequencing (RNA-Seq) and IHC. Molecular and biological effects of silencing RABL6A and/or p27 in MPNST lines and normal human Schwann cells were determined. Tumor-suppressive effects of CDK inhibitors were measured in MPNST cells and orthotopic tumors. RESULTS: RABL6A was dramatically upregulated in human MPNSTs compared with precursor lesions, which correlated inversely with p27 levels. Silencing RABL6A caused MPNST cell death and G1 arrest that coincided with p27 upregulation, CDK downregulation, and RB1 activation. The growth-suppressive effects of RABL6A loss, and its regulation of RB1, were largely rescued by p27 depletion. Importantly, reactivation of RB1 using a CDK4/6 inhibitor (palbociclib) killed MPNST cells in vitro in an RABL6A-dependent manner and suppressed MPNST growth in vivo. Low-dose combination of drugs targeting multiple RB1 kinases (CDK4/6, CDK2) had enhanced antitumorigenic activity associated with potential MPNST cell redifferentiation. CONCLUSIONS: RABL6A is a new driver of MPNST pathogenesis that acts in part through p27-RB1 inactivation. Our results suggest RB1 targeted therapy with multiple pathway drugs may effectively treat MPNSTs.

Threshold Automated Perimetry of the Full Visual Field in Patients With Glaucoma With Mild Visual Loss.

PRECIS: The authors used the Open Perimetry Interface to design a static automated perimetry test of the full field. Abnormal test locations in the nasal midperiphery and temporal inferior sector area best separated glaucomas from normals. PURPOSE: The peripheral visual field in glaucoma outside 30 degrees is largely unexplored with static perimetry. Their goal was to use threshold static automated perimetry to characterize the visual loss in glaucoma of the central 30 degrees and the far periphery. PATIENTS AND METHODS: The authors administered the 30-2 perimetric test to 27 patients with early stage glaucoma (with mean deviation better than -4 dB) with the Goldmann III and V stimulus sizes and a custom test from 30 to up to 87 degrees with the size V stimulus twice within a month. The authors quantified (1) the retest variability, (2) the proportion of patients flagged as abnormal (at level 0.05) on the basis of pointwise probability distributions obtained from 63 ocular healthy observers, (3) the pointwise statistical distance using the Kullback-Leibler divergence between normal and glaucoma eyes, and (4) the effect of eccentricity on visual loss. RESULTS: Size V 30-2 testing identified significantly more abnormal test locations (36%) than size III 30-2 (30%; P=0.004). Kullback-Leibler divergence between healthy and glaucoma distributions was greatest for the nasal midperipheral test locations and the inferior temporal sector area. A more pronounced decrease was found in visual sensitivity with eccentricity in the patients with glaucoma compared with the ocular healthy participants across the full visual field (P<0.001). CONCLUSIONS: Patients with glaucoma demonstrate a systematic decrease in sensitivity with eccentricity across the full visual field. Goldmann size V stimuli better detected visual loss in patients with glaucoma with mild loss than size III.

SITA-Standard perimetry has better performance than FDT2 matrix perimetry for detecting glaucomatous progression.

PURPOSE: The Humphrey Matrix (FDT2) may be more sensitive in detecting glaucomatous visual field loss than SITA standard automated perimetry (SAP) performed on the Humphrey Field Analyzer (HFA). Therefore, FDT may be a good candidate to determine disease progression in patients with glaucoma. Our aim was to test the hypothesis that automated perimetry using the FDT2 would be equal to, or more effective than, HFA SITA-Standard, in identifying glaucomatous progression. METHODS: One hundred and twenty patients with glaucoma were tested twice at baseline and every 6 months for 4 years with HFA SITA-Standard and FDT2. FDT2 values were standardised to HFA SAP values. We used pointwise linear regression (PLR) over the full data series to identify glaucomatous progression and generated an array of results using three different criteria: (1) three or more clustered test locations progressing, (2) three or more non-clustered test locations progressing and (3) total number of progressing test locations. We compared HFA SAP and FDT2 for the number of locations signalled by the PLR detection algorithm. RESULTS: Regardless of the criteria, HFA SAP with SITA-Standard testing detected visual field progression at a higher rate than the FDT2 overall (P<0.001). CONCLUSION: HFA SAP identifies glaucomatous visual field progression at a rate at least as high if not higher than FDT2.

Visual Field Outcomes for the Idiopathic Intracranial Hypertension Treatment Trial (IIHTT).

PURPOSE: The Idiopathic Intracranial Hypertension Treatment Trial (IIHTT) showed that acetazolamide provided a modest, significant improvement in mean deviation (MD). Here, we further analyze visual field changes over the 6-month study period. METHODS: Of 165 subjects with mild visual loss in the IIHTT, 125 had perimetry at baseline and 6 months. We evaluated pointwise linear regression of visual sensitivity versus time to classify test locations in the worst MD (study) eye as improving or not; pointwise changes from baseline to month 6 in decibels; and clinical consensus of change from baseline to 6 months. RESULTS: The average study eye had 36 of 52 test locations with improving sensitivity over 6 months using pointwise linear regression, but differences between the acetazolamide and placebo groups were not significant. Pointwise results mostly improved in both treatment groups with the magnitude of the mean change within groups greatest and statistically significant around the blind spot and the nasal area, especially in the acetazolamide group. The consensus classification of visual field change from baseline to 6 months in the study eye yielded percentages (acetazolamide, placebo) of 7.2% and 17.5% worse, 35.1% and 31.7% with no change, and 56.1% and 50.8% improved; group differences were not statistically significant. CONCLUSIONS: In the IIHTT, compared to the placebo group, the acetazolamide group had a significant pointwise improvement in visual field function, particularly in the nasal and pericecal areas; the latter is likely due to reduction in blind spot size related to improvement in papilledema. (ClinicalTrials.gov number, NCT01003639.).

Older adults catch up to younger adults on a learning and memory task that involves collaborative social interaction.

Learning and memory abilities tend to decline as people age. The current study examines the question of whether a learning situation that emphasises collaborative social interaction might help older persons overcome age-related learning and memory changes and thus perform similarly to younger persons. Younger and Older participants (n = 34 in each group) completed the Barrier Task (BT), a game-like social interaction where partners work together to develop labels for a set of abstract tangrams. Participants were also administered standard clinical neuropsychological measures of memory, on which the Older group showed expected inferiority to the Younger group. On the BT, the Older group performed less well than the Younger group early on, but as the task progressed, the performance of the Older group caught up and became statistically indistinguishable from that of the Younger group. These results can be taken to suggest that a learning milieu characterised by collaborative social interaction can attenuate some of the typical memory disadvantages associated with being older.

Refinement of pointwise linear regression criteria for determining glaucoma progression.

PURPOSE: A variety of pointwise linear regression (PLR) criteria have been proposed for determining glaucomatous visual field progression. However, alternative PLR criteria have only been assessed on a limited basis. The purpose of this study was to evaluate a range of PLR slope and significance criteria to define a clinically useful progression decision rule for longitudinal visual field examinations. METHODS: Visual field data for each of 140 eyes (one per participant among 96 cases and 44 controls) were evaluated using the Humphrey Field Analyzer II program 24-2 Swedish interactive thresholding algorithm standard test strategy and Goldmann size III stimuli. The pointwise linear regression A2 (PLRA2) method was used to analyze the data, which included nine visual field examinations performed every 6 months for 4 years. Data from the Ocular Hypertension Treatment Study (OHTS) were used to validate the decision rule. RESULTS: Several slope criteria produced specificities of 0.90 or higher, particularly slope criteria of less than -1.2 dB/y. The use of the slope criterion less than -1.2 dB/y at a significance level of P < 0.04 for classification resulted in a hit rate of 0.38, more than a 2-fold increase compared with a commonly used standard slope criterion of less than -1.0 dB/y at a significance level of P < 0.01. A similar increase in the hit rate was shown for a slope of less than -1.2 dB/y and P < 0.04 compared with the standard criterion in the independent OHTS validation data. CONCLUSIONS: When systematically evaluating criteria for detecting glaucoma progression, PLR criteria can be refined by requiring a stricter slope criterion such as less than -1.2 dB/y and relaxing the significance criterion to P < 0.04. Increasing the hit rate of PLR will be useful for early detection and treatment of glaucoma.

Size threshold perimetry performs as well as conventional automated perimetry with stimulus sizes III, V, and VI for glaucomatous loss.

PURPOSE: It is thought that large perimetric stimuli are insensitive for demonstrating visual field defects. To test the hypothesis that there is no difference in the total number of abnormal test locations with total deviation empiric probability plots in glaucoma patients, we compared results of glaucoma patients tested with sizes III (0.43° diameter), V (1.72°), and VI (3.44°), and size threshold perimetry (STP), a method that finds threshold by changing stimulus size. METHODS: We derived normative limits for total deviation probability plots using the second test from 60 age-matched normals. We analyzed the probability plots of 120 glaucoma patients (mean deviation was -9.3 ± 6.1 dB with a range of -0.2 to -31.6) at the 42 nonblind spot locations common to the tests. We compared the number of abnormal test locations at the 5% level among the tests using one-way repeated measures ANOVA on ranks. We stratified the results by mean deviation. RESULTS: There was a statistically significant difference in the number of abnormal test locations among the tests: III, 28.5; V, 29.7; VI, 27.0; and STP, 28.8, P = 0.001; Tukey pairwise comparisons were statistically significant for the assessments between sizes V and VI and between STP and size VI. When stratifying by mean deviation, with mild visual loss, size V was most sensitive, followed by STP; size VI appeared slightly less sensitive. CONCLUSIONS: Size V and STP provide favorable stimulus methodology for detection of mild to moderate glaucoma. Size VI appears slightly less sensitive for glaucoma with mild loss.

The repeatability of mean defect with size III and size V standard automated perimetry.

PURPOSE: The mean defect (MD) of the visual field is a global statistical index used to monitor overall visual field change over time. Our goal was to investigate the relationship of MD and its variability for two clinically used strategies (Swedish Interactive Threshold Algorithm [SITA] standard size III and full threshold size V) in glaucoma patients and controls. METHODS: We tested one eye, at random, for 46 glaucoma patients and 28 ocularly healthy subjects with Humphrey program 24-2 SITA standard for size III and full threshold for size V each five times over a 5-week period. The standard deviation of MD was regressed against the MD for the five repeated tests, and quantile regression was used to show the relationship of variability and MD. A Wilcoxon test was used to compare the standard deviations of the two testing methods following quantile regression. RESULTS: Both types of regression analysis showed increasing variability with increasing visual field damage. Quantile regression showed modestly smaller MD confidence limits. There was a 15% decrease in SD with size V in glaucoma patients (P = 0.10) and a 12% decrease in ocularly healthy subjects (P = 0.08). CONCLUSIONS: The repeatability of size V MD appears to be slightly better than size III SITA testing. When using MD to determine visual field progression, a change of 1.5 to 4 decibels (dB) is needed to be outside the normal 95% confidence limits, depending on the size of the stimulus and the amount of visual field damage.

Modeling lateral control in driving studies.

In driving studies based on simulators and instrumented vehicles, specific models are needed to capture key aspects of driving data such as lateral control. We propose a model that uses weighted polynomial projections to predict each data point from the previous three time points, and accommodates the attempts of the drivers to re-center the vehicle before crossing the borders of the traffic lane. Our model also allows the possibility that average position within the lane may vary from driver to driver. We demonstrate how to fit the model using standard statistical procedures available in software packages such as SAS. We used a fixed-base driving simulator to obtain data from 67 drivers with Alzheimer's disease and 128 elderly drivers without dementia. Using these data, we estimated the subject-specific parameters of our model, and we compared the two groups with respect to these parameters. We found that the parameters based on our model were able to distinguish between the groups in an interpretable manner. Hence, this model may be a useful tool to define outcome measures for observational and interventional driving studies.

Pharmacologic modification to resuscitation fluid after thermal injury--is drotrecogin alfa the answer to arrest burn depth progression?

HYPOTHESIS: The addition of drotrecogin alfa (DA), an anti-inflammatory useful in septic shock, to standard burn shock resuscitation fluids will protect burned, injured skin from further injury. METHODS: Anesthetized animals were subjected to a standardized burn pattern by applying a branding iron to 10 different locations on the back of the rat for 1 seconds to 14 seconds, creating a range of burn depths and severities. DESIGN: Animal burn shock and resuscitation model. PARTICIPANTS: Thirty-one male adult Sprague-Dawley rats. INTERVENTIONS: Control animals were resuscitated with lactated Ringer's solution (LRS) at 2 mL/kg/percent total body surface area/24 h; experimental animals received LRS plus DA 24 microg/kg/h (LRS + DA). OUTCOME MEASURES: Perfusion to each burned area was assessed using a laser Doppler imaging technology. Punch biopsies at each burned area were stained with hematoxylin and eosin and assessed for burn depth and for inflammation using previously reported measures. Samples from 14 animals were stained for terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and caspase-3 (apoptosis markers). RESULTS: Increasing branding iron contact times worsened perfusion, burn depth, and apoptotic ratios. There was no correlation between inflammatory markers and burn contact time. The addition of DA leads to worse perfusion, deeper burns, worse inflammation, and decreased apoptotic ratios. CONCLUSIONS: Laser Doppler imaging is a useful technology to assess burn depth. The addition of DA to traditional resuscitation fluids for burn shock is deleterious to the injured, burned skin. Modifying the traditional burn shock resuscitation fluids, although intellectually attractive, needs to be rigorously studied.

Liquid ventilation with perfluorocarbons facilitates resumption of spontaneous circulation in a swine cardiac arrest model.

BACKGROUND: Induced external hypothermia during ventricular fibrillation (VF) improves resuscitation outcomes. Our objectives were twofold (1) to determine if very rapid hypothermia could be achieved by intrapulmonary administration of cold perfluorocarbons (PFC), thereby using the lungs as a vehicle for targeted cardiopulmonary hypothermia, and (2) to determine if this improved resuscitation success. METHODS: Part 1: Nine female swine underwent static intrapulmonary instillation of cold perfluorocarbons (PFC) during electrically induced VF. Part 2: Thirty-three female swine in VF were immediately ventilated via total liquid ventilation (TLV) with pre-oxygenated cold PFC (-15 degrees C) or warm PFC (33 degrees C), while control swine received no ventilation during VF. All swine in both Parts 1 and 2 underwent VF arrest for 11 min, then defibrillation, ventilation and closed chest massage until resumption of spontaneous circulation (ROSC). The endpoint was continued spontaneous circulation for 1h without pharmacologic support. RESULTS: Static intrapulmonary instillation of cold PFC achieved rapid cardiopulmonary hypothermia; pulmonary artery (PA) temperature of 33.5+/-0.2 degrees C was achieved by 10 min. Nine of 9 achieved ROSC. Hypothermia was achieved faster using TLV: at 6 min VF, cold TLV temperature was 32.9+/-0.4 degrees C vs. cold static instillation temperature 34.3+/-0.2 degrees C. Nine of 11 cold TLV swine achieved ROSC for 1h vs. 3 of 11 control swine (p=0.03). Warm PFC also appeared to be beneficial, with a trend toward greater achievement of ROSC than control (ROSC; warm PFC 8 of 11 vs. control 3 of 11, p=0.09). CONCLUSION: Targeted cardiopulmonary intra-arrest moderate hypothermia was achieved rapidly by static intrapulmonary administration of cold PFC and more rapidly by total liquid ventilation with cold PFC; resumption of spontaneous circulation was facilitated. Warm PFC showed a trend toward facilitating ROSC.