Bivalent target-binding bioPROTACs induce potent degradation of oncogenic SHP2.

Targeted protein degradation is an emergent and rapidly evolving therapeutic strategy. In particular, biologics-based targeted degradation modalities (bioPROTACs) are relatively under explored compared to small molecules. Here, we investigate how target affinity, cellular localization, and valency of bioPROTACs impact efficacy of targeted degradation of the oncogenic phosphatase src-homology 2 containing protein tyrosine phosphatase-2 (SHP2). We identify bivalent recruitment of SHP2 by bioPROTACs as a broadly applicable strategy to improve potency. Moreover, we demonstrate that SHP2-targeted bioPROTACs can effectively counteract gain-of-function SHP2 mutants present in cancer, which are otherwise challenging to selectively target with small molecule constructs. Overall, this study demonstrates the utility of bioPROTACs for challenging targets, and further explicates design principles for therapeutic bioPROTACs.

Comparative Effectiveness of Hybrid Ablation Versus Endocardial Catheter Ablation Alone in Patients With Persistent Atrial Fibrillation.

OBJECTIVES: The outcomes of hybrid ablation versus endocardial catheter ablation alone were evaluated in patients with persistent and long-standing persistent atrial fibrillation (AF). BACKGROUND: Variable outcomes exist following endocardial catheter ablation in medically refractory patients with persistent AF. A hybrid epicardial-endocardial approach has emerged as an alternative to endocardial ablation. METHODS: In 133 consecutive patients, 69 received endocardial ablation alone (pulmonary vein isolation and radiofrequency catheter ablation [endo group]) and 64 received endocardial catheter ablation and epicardial ablation (hybrid group). Recurrence was defined as any arrhythmia following the 3-month blanking period. RESULTS: Patients were followed for a median of 16 months. The hybrid and endo groups were similar in age (61 ± 10 years vs. 62 ± 8 years), body mass index (35 ± 6 kg/m2 vs. 35 ± 7 kg/m2), CHA2D2-VASc score (2 ± 1 vs. 2 ± 1), and ejection fraction (54 ± 11% vs. 53 ± 8%). The hybrid group had longer AF duration (median [interquartile range (IQR)] (12 months [IQR: 8 to 28 months] vs. 7 months [IQR: 5 to 12 months]; p < 0.001) and more previous ablations (58% vs. 25%; p < 0.001). Both groups had similar antiarrhythmic drug use at follow-up (55% vs. 48%). The hybrid group was less likely to have recurrence (37% vs. 58%; p = 0.013) and repeat ablation (9% vs. 26%; p = 0.012), and had an AF-free survival of 72% versus 51% (p = 0.01). CONCLUSIONS: Among patients with persistent AF, hybrid ablation is associated with less AF recurrence and fewer re-do ablations. Prospective large-scale randomized trials are needed to validate these results.

A randomized, prospective trial evaluating surgeon preference in selection of absorbable suture material.

This study is the first double-blinded, randomized comparison of two absorbable sutures. To better understand product characteristics and surgeon preference, we conducted a study of two similar-appearing FDA-approved sutures, glyconate and poliglecaprone 25. Four dermatologic surgeons were enlisted. A total of 48 patients with 53 surgical sites were examined. One half of each surgical wound was closed with one type of suture and the other half with the other type. Each half was evaluated for product characteristics. There was no statistically significant difference in surgeon preference for glyconate versus poliglecaprone 25 (P=0.64). Of the cohort preferring poliglecaprone 25, there was a correlation with speed of closure (P=0.06). Of the surgeons that preferred glyconate, we found significantly better visibility (P=0.03), reduced suture breakage during knot tying (P=0.05), and correlation with better handling properties (P=0.06) associated with that preference. The data from this study will enable products to be designed towards these needs and allow surgeons to select sutures that more precisely fit their particular requirements.

Computed tomography-fluoroscopy image integration-guided catheter ablation of atrial fibrillation.

INTRODUCTION: This study examines the feasibility of atrial fibrillation (AF) ablation using registered three-dimensional computed tomography (CT) images of the left atrium with fluoroscopy. METHODS AND RESULTS: A total of 50 consecutive patients with symptomatic AF refractory to medical therapy (32 paroxysmal, 18 persistent, age 55 +/- 10 years) were randomized to undergo a catheter-based AF ablation procedure with or without the CT-fluoroscopy guidance system. All patients underwent preprocedural contrast-enhanced CT imaging and segmentation of the left atrium. For the CT-fluoroscopy group, circumferential lesions encompassing the pulmonary vein (PV) antrum and linear lesions along the roof of the left atrium between the superior PVs and the mitral isthmus were created on the CT image, which was registered with real-time fluoroscopy. The registered images were then used to navigate the ablation catheters to the sites of planned ablation. After the ablation sites were completed, any remaining PV potentials were isolated with electrophysiological guidance. In the control patients, the same technique was performed without using the CT-fluoro guidance system. CT scans were accurately registered to fluoroscopic images with minimal manual correction. Operators could navigate catheters on the registered images to preplanned, extraostial sites for ablation. CT-fluoroscopy guidance decreased procedure duration and fluoro times (P < 0.05). At a mean follow-up of 9 +/- 2 months, 21 patients (84%) in the CT-fluoro guidance group and 16 patients (64%) in the control group have had no recurrence of AF. CONCLUSION: CT-fluoroscopic-guided left atrial ablation is feasible and allows appropriate catheter manipulation in the left atrium.

Effect of electrical stimulation on arteriogenesis and angiogenesis after bilateral femoral artery excision in the rabbit hind-limb ischemia model.

The effects of electrical stimulation (ES) on arteriogenesis (the opening of preexisting collaterals) and angiogenesis (formation of new capillaries) were studied after acute bilateral hind limb ischemia was induced via bilateral femoral artery excision in a rabbit model. The study evaluated the rabbit hind limbs' normal response to acute ischemia and to application of ES by calculating changes in arterial and capillary densities. Comparisons were made with our prior study, in which the femoral artery was unilaterally excised, as we attempted to expand on the topics of arteriogenesis and angiogenesis. Twelve adult New Zealand white rabbits were randomly assigned to 1 of 2 series. In Series 1, the control group, both femoral arteries were excised and no ES was applied. In Series 2, both femoral arteries were excised and ES was applied to the left limb. One lead was implanted into the left adductor muscle near the site of the excised left femoral artery (Series 2), and a stimulator (Thera, Medtronic, Inc, Minneapolis, MN) was implanted in a separate pocket. ES was applied at a rate of 3 V, 30 contractions per minute, beginning immediately after surgery and continuously for 1 month. Angiography was performed in all 12 rabbits 1 month after surgery to establish the anatomy of the collateral vessels and to demonstrate that the femoral artery stump continued to be an end artery. Contrast-opacified arteries (COAs) that crossed the grid's midline, and the total number of grid lines intersected by COAs, were tallied according to an established method. Capillary density was calculated as the number of capillaries per square millimeter of muscle. In Series 1, after 1 month, the number of COAs crossing the grid's midline was 4.5 +/-1.5 on the left and 4.8 +/-1.2 on the right side. In Series 2, the number of COAs crossing the grid's midline was 7.9 +/-1.8 on the left side (p<0.05 vs Series 1) and 5.9 +/-1.6 on the right side of the same rabbit (p=NS vs Series 1). In Series 1, 36.7 +/-5.4 and 30.5 +/-7.7 total intersections were crossed by COAs on the left and right sides, respectively. In Series 2, total grid intersections crossed by COAs were 48.4 +/-8.5 and 47.5 +/-9.1 in the left and right sides, respectively (p<0.001 vs series 1). Baseline capillary density before femoral artery excision was 180.2 +/-21.3/mm(2). The capillary densities on the left sides were 94.2 +/-19.1 and 264.5 +/-7.6 in Series 1 and 2, respectively (p<0.001). The right sides showed a similar pattern with capillary densities of 88.5 +/-37.2 and 135.8 +/-6.8 (p<0.05) in Series 1 and 2, respectively. When capillary density was compared on the left and right sides of the same rabbit in Series 2, a statistically significant increase was also found; 264.5 +/-7.6 vs 135.8 +/-6.8 (p<0.001) in the left and right sides, respectively. Comparisons of the effect of electrical stimulation and the body's normal physiologic response to acute ischemia revealed a significant increase in the opening of preexisting collaterals (arteriogenesis) and the promotion of capillary density (angiogenesis) with the use of electrical stimulation.

Slowed progression or elimination of atherosclerosis by low-frequency electrical impulses.

BACKGROUND: In our previous investigations we showed that electrical impulses (EI) can prevent the development of atherosclerosis if they began simultaneously with high cholesterol diet (HCD) or in the early stages of atherosclerosis (after three weeks of HCD only). In this investigation we demonstrated the slow progression or elimination of atherosclerosis by low-frequency EI in case of moderate atherosclerosis (after eight weeks of HCD). METHODS: Series I rabbits (control group) were fed HCD for eight weeks. Series II rabbits were fed HCD for eight weeks and were then switched to normal diet for eight weeks (no EI). Series III rabbits were fed HCD for eight weeks and then switched to a normal diet with simultaneous EI (applied near the abdominal aorta) for eight weeks (3 V, 30 single impulses per minute, 24 hours/day). After euthanization, the level of atherosclerosis, percentage of surface area involved in the atherosclerosis process, and an atherosclerosis score were calculated in the aortic arch, thoracic and abdominal aorta. RESULTS: Statistically significant differences were seen in the level of atherosclerosis in the abdominal aorta between series III animals (0.4 +/- 0.2) and the other two groups: 1.5 +/- 0.4 in series I (HCD only), 1.2 +/- 0.3 in series II (HCD then normal diet). Gross examination of the surface also revealed statistically significant differences (p < 0.05) in the percentage of atherosclerosis between the control series I (30.1 +/- 4.1%) and series II (21.3 +/- 3.6%), compared with series III (5.5 +/- 5.4%). In addition, the atherosclerosis score was also significantly different: 45.8 +/- 3.9 in series I, 25.2 +/- 6.9 in series II, and 2.2 +/- 2.0 in series III (p < 0.05). CONCLUSION: Our study showed that, when applied near the abdominal aorta, low-frequency electrical impulses decrease atherosclerotic deposition in the abdominal aorta.

Electrical stimulation promotes angiogenesis in a rabbit hind-limb ischemia model.

In previous investigations, it was shown that applying a modest regimen of electrical stimulation (ES), even in severely ischemic tissue, improves the healing process, accelerates neovascularization, and enhances angiogenesis in muscle tissue. Our objective in this current report was to further understand ES as a potential alternative treatment for severe muscle ischemia. Immediately after the left distal external iliac artery and the femoral artery were excised, ES (30 contractions per minute [cpm], 2 V, single impulses per burst) was applied to rabbit adductor muscle near the site of the excised femoralis artery for 24 hours daily over 1 month. Three other series served as controls: ES without arterial excision; arterial excision without ES or lead implantation; and arterial excision with lead implantation but no ES. Histologic study of capillary density was performed by angiography (employing a grid template) and by measuring the lower limb-calf blood pressure ratio. At the end of 30 days in the ES series, 10.5 +/-1.2 contrast-medium opacified arteries (COAs) crossed a specific grid section segment compared with 7.2 +/-1.5 in the control series without ES (p<0.05); 68.2 +/-9.3 COAs crossed a grid section compared with 43.2 +/-6.4 in controls (p<0.05); 27.3 +/-1.2 grids contained COAs compared with 29.3 +/-3.5 in controls (p<0.05); lower limb-calf blood pressure ratio was 0.81 +/-0.06 compared with 0.31 +/-0.07 in controls (p<0.05); and capillary density was 283.7 +/-24.5 mm2 compared with 91.4 +/-20.9 mm2 in controls (p<0.001). These preliminary results show that cautious ES enhances and accelerates muscle revascularization in severely ischemic tissue.

Radiofrequency ablation of atrial fibrillation during mitral valve surgery.

Twenty-three patients underwent endocardial radiofrequency ablation of atrial fibrillation (AF) during mitral valve procedures with a previously described left atrial lesion pattern. A temperature-controlled 7-coil surgical probe delivered 60-second lesions at 80 degrees C. The left atrial appendage was oversewn after ablation. Ages ranged from 28 to 88 years. Nineteen patients had chronic AF that was present for over 1 year in 74%; 12 patients had rheumatic mitral stenosis. Mean left atrial diameter was 5.4 +/- 0.7 cm. There was 1 operative death unrelated to the ablation, and no strokes or ablation-related complications were observed. At mean follow-up of 32.5 weeks, 86% of the 22 survivors were in sinus rhythm. All 18 patients with left atrial diameter <6 cm are in sinus rhythm. All postoperative atrial flutter was transient, and no patients required subsequent transcatheter ablation. This lesion pattern is safe and effective when applied in the method described here. It appears to be a reasonable alternative to the complete Maze 3 lesion pattern in patients with mitral valve disease.

Validation of a left atrial lesion pattern for intraoperative ablation of atrial fibrillation.

BACKGROUND: Evidence that atrial fibrillation may begin in early stages from triggers or reentry circuits primarily in the left atrium suggests that the entire Maze 3 lesion pattern may be unnecessary. In the present study we describe a new left atrial lesion pattern for intraoperative linear ablation of chronic atrial fibrillation. METHODS: Endocardial radiofrequency ablation was performed on 12 dogs with chronic atrial fibrillation. Lesions to isolate pulmonary veins in pairs, the left atrial appendage, and connecting lesions between these structures were administered in a randomized approach. RESULTS: Twelve dogs were in chronic atrial fibrillation for 31 +/- 21 days before ablation. Atrial fibrillation was successfully ablated and rendered noninducible in all 12 dogs. All treatment failures observed with less than the full lesion pattern became a success when the remaining lesions were given. CONCLUSIONS: Atrial fibrillation ablation using this left atrial lesion pattern is highly successful in this model. This approach may have significant utility as a concomitant procedure for patients with atrial fibrillation undergoing mitral valve procedures.

Pathologic and histologic results of electrical impulses in a rabbit model of atherosclerosis: 24-hour versus 8-hour regimen.

OBJECTIVE: Low frequency electrical impulses (EIs) reduce new atherosclerotic plaque formation in previously diseased arteries and may reverse the extent of previous pathologic damage in these structures. METHODS: A pacemaker was implanted on the left side of rabbit abdominal aortas, and an electrode was placed close to the other side of the aorta in the psoas major muscle. For the induction of atherosclerosis, the rabbits were placed on a high cholesterol diet (HCD) for 11 weeks. No EIs were applied to the control series I. In the experimental series, the rabbits were fed an HCD for 3 weeks, after which EIs were applied simultaneously with an HCD for 8 additional weeks (3V, 30 contractions per minute). Experimental series II had 24-hour/day EIs, and series III had 8-hour/day EIs. RESULTS: The closer to the area where the EIs were applied, the more local severity increased (atherosclerosis level and surface area). In the control series, the severity of atherosclerosis in the lower aorta assessed with an arbitrary grading system was 1.75 +/- 0.5 (versus 1.5 +/- 0.57 with 8-hour/day EIs and 0.5 +/- 0.3 with 24-hour/day EIs). The involved surface area was 32.5% +/- 9.5% (versus 1.0% +/- 0.8% with 8-hour/day EIs and 0.75% +/- 0.95% with 24-hour/day EIs). CONCLUSION: Both 24-hour/day and 8-hour/day EIs applied close to the abdominal aorta decreased the severity of atherosclerosis in rabbits placed on a HCD, but 24-hour/day EIs decreased the severity more extensively.