Search

Objective Transcarotid access for transcatheter aortic valve replacement is emerging as an alternative to more traditional nonfemoral access options such as transapical or transaortic; however, comparative data are limited. The purpose of the study was to analyze outcomes after transcatheter aortic valve replacement using transcatheter compared with transthoracic (transapical/transaortic) access. Methods The Society of Thoracic Surgeons/American College of Cardiology Transcatheter Valve Therapy Registry was queried for patients who underwent transcarotid, transapical, or transaortic transcatheter aortic valve replacement with the SAPIEN 3 (Edwards Lifesciences, Irvine, Calif) transcatheter heart valve between June 2015 and July 2019. Thirty-day unadjusted outcomes were evaluated, and propensity score matching and logistic regression were used to compare transcatheter access with transthoracic access. Results In the propensity-matched analysis, 667 transcarotid transcatheter aortic valve replacement procedures were compared with 1334 transthoracic procedures. Transcarotid transcatheter aortic valve replacement was associated with lower mortality (4.2% vs 7.7%, P = .004), less new-onset atrial fibrillation (2.2% vs 12.1%, P Conclusions Transcatheter aortic valve replacement using transcarotid access is associated with lower 30-day mortality, less atrial fibrillation, shorter intensive care unit and overall length of stay, fewer readmissions, greater improvement in Kansas City Cardiomyopathy Questionnaire scores, and no significant difference in stroke or major vascular complications compared with transthoracic access.

Supplementary Figure 2. (A) Quantification of relative fluorescence intensity units (RFU) of ß1 integrin in BT459 and MDA-MB231LN cells as in (B); graph is mean RFU, % of siCon (assigned as 100%) +/-S.E.M; n=3, 15 cells/per treatment; one-way ANOVA with Dunnett's post-hoc analysis; ligand-bound: MDA-MD-231LN *p

Representative movie of individual MDA-MB-231LN cells treated with Vehicle, MLN8237, eribulin, or MLN8237/eribulin combination, suspended in collagen, and manually tracked over the course of 23 hours toward the serum chemoattractant-containing reservoir (left side of frame).

Ibidi chemotactic invasion of MDA-MB-231 cells with CA-RhoA overexpression

File contains Supplementary figures: Figure 1: AUKRA analysis in TNBC cell lines in response to drug treatments. Additional TNBC migration/invasion analysis. Figure 2. Representative FACS analysis flow charts of TNBCs undergoing apoptosis. Figure 3. F-IHC analysis of tumors and lung metastases at the end point of treatment with vehicle, MLN8237, eribulin with indicated phospho-specific antibodies. Figure 4. Representative images of F-IHC, DNA (DAPI) staining of pulmonary tissue (metastases defined by the white outline) and mammary tumors. Figure 5. Fluorescent IHC analysis of metastases using pan-cytokeratin antibodies (Dako, green) to detect human tumor cells in lungs (mouse), DNA-DAPI. Figure 6. WB analysis of LC3BI/II, pAKT, AKT, alpha-tubulin expression in tumor cells isolated mammary tumors and lungs. Figure 7. F-IHC analysis of mammary tumor and lung metastases cells with p62, pAKT antibodies in the presence of Chloroquine. Figure 8. F-IHC staining of pulmonary tissue from, -cleaved Caspase-3, -phosphoAKT1/2. Figure 9. Cell growth analysis of Tumor cells PEN65 derived from PDX treated with indicated drugs. Figure 10. LAESI-MS MS/MS fragmentation of MLN8237 in dosed animal tumor tissue.

Commonly upregulated in human cancers, the scaffolding protein NEDD9/HEF1 is a known regulator of mesenchymal migration and cancer cell plasticity. However, the functional role of NEDD9 as a regulator of different migration/invasion modes in the context of breast cancer metastasis is currently unknown. Here, it is reported that NEDD9 is necessary for both mesenchymal and amoeboid individual cell migration/invasion in triple-negative breast cancer (TNBC). NEDD9 deficiency results in acquisition of the amoeboid morphology, but severely limits all types of cell motility. Mechanistically, NEDD9 promotes mesenchymal migration via VAV2-dependent Rac1 activation, and depletion of VAV2 impairs the ability of NEDD9 to activate Rac1. In addition, NEDD9 supports a mesenchymal phenotype through stimulating polymerization of actin via promoting CTTN phosphorylation in an AURKA-dependent manner. Interestingly, an increase in RhoA activity in NEDD9-depleted cells does not facilitate a switch to functional amoeboid motility, indicating a role of NEDD9 in the regulation of downstream RhoA signaling effectors. Simultaneous depletion of NEDD9 or inhibition of AURKA in combination with inhibition of the amoeboid driver ROCK results in an additional decrease in cancer cell migration/invasion. Finally, we confirmed that a dual targeting strategy is a viable and efficient therapeutic approach to hinder the metastasis of breast cancer in xenograft models, showcasing the important need for further clinical evaluation of this regimen to impede the spread of disease and improve patient survival.Implications: This study provides new insight into the therapeutic benefit of combining NEDD9 depletion with ROCK inhibition to reduce tumor cell dissemination and discovers a new regulatory role of NEDD9 in the modulation of VAV2-dependent activation of Rac1 and actin polymerization. Mol Cancer Res; 15(6); 670–82. ©2017 AACR.

Supplementary Figure 3. (A). Quantification of volume of early endosomes based on 3D reconstructed confocal images of BT459-siCon, -siN2, -siN3) cells stained with EEA1 antibodies; mean volume (%) normalized to siCon+S.E.M; n=3, 10 cells/treatment; one-way ANOVA: *p

File contains detailed information on Supplementary video files.

Table contains diagnostics/characterization of original and patient-derived models.

File contains detailed protocols and information on used reagents to facilitate reproducible of the presented data.

Representative movie of individual BT-549 cells treated with Vehicle, MLN8237, eribulin, or MLN8237/eribulin combination, suspended in collagen, and manually tracked over the course of 23 hours toward the serum chemoattractant-containing reservoir (left side of frame).

Fig. S1. NEDD9 depletion causes a switch to amoeboid morphology. Fig. S2. NEDD9 depletion decreases phosphorylation of focal adhesion proteins in other BC cell lines. Fig. S3. NEDD9 depletion affects vinculin-containing focal adhesion dynamics. Fig. S4. NEDD9 depletion affects talin-containing focal adhesion dynamics. Fig. S5. AURKA phosphorylates CTTN in breast cancer cells. Fig. S6. NEDD9 depletion decreases Rac1 activity in other breast cancer cell lines. Fig. S7. Depletion of NEDD9 via CRISPR-Cas9 system and overexpression of CA-RhoA decreases cell invasion and increases round amoeboid phenotype. Fig. S8. Simultaneous inhibition of mesenchymal and amoeboid pathways through NEDD9 and RhoA knockdown hinders cell invasion in vitro. Fig. S9. Depletion of NEDD9 hinders invasion of other breast cancer cell lines in vitro. Fig. S10. AURKA inhibition hinders breast cancer cell invasion.

Supplementary Figure 1. (A). ECIS-BT549-shCon or shNEDD9 (N1, N4) cells, 5h of adhesion/spreading ;quantification of electrical impedance normalized to control +/-S.E.M; n=4; one way ANOVA: *p< 0.0157 shCon/N1or N4; (B). Boyden chamber migration assay with MDA-MB-231 cells treated with siCon or siN2 for 8h, quantification of RFU (% to siCon assigned as 100%) +/-S.E.M in ; n=3, * p

The diverse structures and profound biological activities of lignan natural products have enticed significant effort in the exploration of new methodologies for their total synthesis. We have prepared γ-butyrolactone oximes from readily available δ-nitro alcohols via Boc(2)O mediated cyclization. The mild conditions are compatible with a wide range of functional groups, and this methodology has been applied to the total synthesis of five lignan natural products.

There are limited invasive treatment options for patients with end-stage heart failure and left ventricular assist device (LVAD) who develop severe aortic valve regurgitation (AR). One option for such patients is transcatheter aortic valve replacement (TAVR). There are limited data on outcomes of patients with LVAD who receive TAVR for severe AR. We present a series of 4 consecutive patients with LVAD who underwent TAVR for severe AR.This is a retrospective chart review of 4 consecutive patients with LVAD who underwent TAVR for severe AR. All 4 patients underwent TAVR with a 34-mm self-expanding valve (Medtronic). One patient received a 29-mm balloon-expandable valve (Edwards Lifesciences) within the self-expanding valve (SEV) to postdilate the SEV and minimize paravalvular leak (PVL). All 4 procedures were technically successful. The patient who received rescue valve-in-valve TAVR continued to have persistent mild to moderate PVL.Although technically challenging, TAVR is a feasible option for carefully selected LVAD patients with severe AR. Procedural issues to consider include oversizing the transcatheter heart valve (THV) while being cognizant of the risks of annular rupture and valve dislocation, anticipating and avoiding ventricular migration of the THV and being ready to postdilate the THV if necessary, to limit hemodynamically significant PVL.

Estrogen receptor-positive breast cancers constitute the majority of newly diagnosed breast cancers in patients. While commonly treated with various approved endocrine therapy strategies, these cancers can unfortunately also develop resistance to such therapies, indicating a need to explore other avenues of treatment in endocrine-resistant breast cancer. Rapidly dividing cancer cells are known to have an increased energy demand and altered reprogramming of glucose metabolism, leading to a subsequent heavy reliance on glycolysis. A critical rate-limiting step in the glycolysis pathway, the conversion of fructose-6-phosphate to fructose-1,6-bisphosphate, is greatly influenced by an allosteric activator produced by the enzyme PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3), thus making PFKFB3 an essential regulator of glycolytic flux. Furthermore, PFKFB3 has been shown to be overexpressed in many human cancers, including breast cancer, as well as be upregulated by estrogen. Through the following work, we show that endocrine-resistant BC cell growth is strongly reduced by PFKFB3 targeting using different commercial PFKFB3 inhibitors, which can be further exacerbated by combination treatment with well-established endocrine therapies like fulvestrant in some cell lines. Moreover, we relatedly show that PFKFB3 inhibition reduced tumor size in mouse xenografts of endocrine-resistant BC cells. Endocrine-resistant cells also presented higher basal glucose uptake and greater reduction of glucose uptake in response to PFKFB3 inhibition than endocrine-sensitive cells, indicating that endocrine-resistant breast cancer cells may have a more significant reliance on glycolysis and PFKFB3 activity for their metabolic needs than sensitive cells. Additionally, PFKFB3 inhibition produced an increase in phosphorylation of the necroptotic markers RIPK1 (receptor-interacting kinase 1), and MLKL (Mixed Lineage Kinase Domain Like Pseudokinase), and pre-treatment with the necroptosis inhibitor necrostatin-1 was able to partially rescue cell death caused by PFKFB3 targeting, suggesting that necroptosis is a primary method of cell death induced by PFKFB3 inhibition. In summation, this study showcases the glycolytic enzyme PFKFB3 as an encouraging therapeutic target in endocrine-resistant breast cancer, and warrants further review as a monotherapy or in combination with current endocrine therapies. Citation Format: Brandon C Jones, Surojeet Sengupta, Catherine M Sevigny, Lu Jin, Paula R Pohlmann, Ayesha Shajahan-Haq, Robert Clarke. Pfkfb3 inhibition significantly decreases endocrine-resistant breast cancer growth and induces necroptotic cell death [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-02-10.