Administrative supplement - Equipment

行政补充-设备

• 批准号: 10378986
• 负责人: Flordeliza S Villanueva
• 金额: $ 3.57万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378986
• 关键词: Acoustics; Address; Administrative Supplement; Automobile Driving; Behavior; Biological Response Modifier Therapy; Biology; Blood Vessels; Cardiovascular Diseases; Cell membrane; Cells; Confocal Microscopy; Contracts; Custom; DNA; Development; Disease; Endothelial Cells; Endothelium; Equipment; Event; Extravasation; Gene Delivery; In Vitro; Intravenous; Knowledge; Life; Malignant Neoplasms; Measures; Mechanics; Mediating; Methods; Microbubbles; Microcirculation; Molecular; Nucleic Acids; Optics; Pathologic; Pathway interactions; Permeability; Physics; Physiology; Property; Proteins; Research; Savings; Signal Transduction; Site; Small Interfering RNA; Speed; Technology; Testing; Therapeutic; Time; Translations; Ultrasonic Transducer; Ultrasonography; experimental study; gene therapy; image guided; in vivo; insight; macromolecule; multidisciplinary; nucleic acid delivery; nucleic acid-based therapeutics; real-time images; response; sound; targeted delivery; vibration

Increasing  knowledge  of  the  molecular  underpinnings  of  disease  is  driving  a  powerful  imperative  to  deliver  agents, such as nucleic acids, to silence expression of pathologic proteins for life-­saving treatment of heretofore  hopeless  illnesses.  Although  there  are  promising  developments  in  strategies  to  deliver  cell  membrane  impermeant  nucleic  acids,  such  as  siRNA,  to  disease-­causing  cells,  a  safe  and  efficient  method  for  targeted  delivery of these agents has remained elusive. A significant hurdle for gene therapies using vascular delivery is  to circumvent the endothelial barrier. We have been developing a unique technology using intravenously injected  nucleic acid-­loaded microbubbles (MB) which are triggered to cavitate (expand and contract) by ultrasound (US),  causing transient permeabilization of the adjacent cell membrane and delivery of the therapeutic carried by the  MBs. The potential of this site-­specific, non-­invasive delivery method is extraordinary, more so because the MBs  and US transducer also provide capability for simultaneous real time image-­guided navigation of therapy. Despite  its promise, the mechanisms underlying the efficacy of ultrasound-­triggered MB cavitation (UTMC) as a delivery  platform are poorly understood. Without a sound knowledge of the fundamental mechanisms by which safe and  effective  biotherapeutic  delivery  is  effected  by  UTMC,  its  ultimate  bedside  translation  is  impossible.  We  hypothesize that MBs cavitating in the microcirculation impart non-­lethal mechanical perturbations on endothelial  cells,  leading  to  signaling  events  that  culminate  in  endothelial  barrier  hyperpermeability.  We  propose  in  vitro  studies to systematically interrogate mechanistic pathways, followed by in vivo experiments to investigate UTMC  endothelial  barrier  effects  in  real  time,  addressing  three  Specific  Aims:  (1)  Determine  the  mechanisms  by  which UTMC increases endothelial barrier permeability. We will use transwells coated with endothelial cells  and manipulate candidate pathways to test the hypothesis that UTMC-­induced Ca2+ influx increases endothelial  permeability.  We  will  optically  measure  attendant  cellular  events  using  multicolor  confocal  microscopy,  thus  correlating  barrier  function  to  cell  response;;  (2)  Determine  the  relationship  between  in  vivo  MB  cavitation  behaviors  and  transendothelial  transport  of  macromolecules  (siRNA).  We  will  use  a  custom  ultra-­high  speed  camera  to  visualize  in  vivo  US-­MB  vibrations  in  the  microcirculation  to  test  the  hypothesis  that  MB  cavitation  causes  quantifiable  mechanical  events,  then  derive  physical  principles  governing  UTMC-­mediated  hyperpermeability;; (3): Determine extravasation pathways and cellular fate of siRNA-­loaded MBs during  UTMC in vivo. We will use intravital high-­speed multicolor confocal microscopy in cremaster microcirculation to  visualize  endothelial  barrier  properties  and  siRNA-­loaded  MB  fate.  Our  multi-­disciplinary  team  unites  physics/acoustics with biology/physiology to derive insights into fundamental physical and cellular mechanisms  underlying UTMC-­facilitated gene delivery. Ultimately, our research will define a rational basis for optimization  of this remarkable technology and accelerate the translation of nucleic acid therapeutics to the bedside.

对疾病的分子基础知识的不断增加，推动了实现这一目标的强烈要求

项目成果

Complex Highways on the Translational Roadmap for Therapeutic Ultrasound-Targeted Microbubble Cavitation: Where Are We Now?

超声靶向微泡空化治疗转化路线图上的复杂高速公路：我们现在在哪里？

• DOI: 10.1016/j.jcmg.2019.08.010
• 发表时间: 2020
• 期刊: JACC. Cardiovascular imaging
• 作者: Villanueva,FlordelizaS;Chen,Xucai
• 通讯作者: Chen,Xucai

A multicellular brain spheroid model for studying the mechanisms and bioeffects of ultrasound-enhanced drug penetration beyond the blood‒brain barrier.

• DOI: 10.1038/s41598-023-50203-3
• 发表时间: 2024-01-22
• 期刊: Scientific reports
• 影响因子: 4.6

Ultrafast Microscopy Imaging of Acoustic Cluster Therapy Bubbles: Activation and Oscillation.

声簇治疗气泡的超快显微成像：激活和振荡。

• DOI: 10.1016/j.ultrasmedbio.2022.05.009
• 发表时间: 2022
• 期刊: Ultrasound in medicine & biology
• 影响因子: 2.9
• 作者: vanWamel,Annemieke;Mühlenpfordt,Melina;Hansen,Rune;Healey,Andrew;Villanueva,FlordelizaS;Kotopoulis,Spiros;Davies,CatharinadeLange;Chen,Xucai
• 通讯作者: Chen,Xucai