Notification of abstract acceptance
announced via E-mail on July 31st.
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Oct. 19 (Fri)  201 Room - WN
08:45-09:15 [WN-03] 
Sonoporation: preclinical application
   
Speaker Jae Young Lee (Seoul National University College of Medicine, Korea.)
Authors Jae Young Lee
Affiliation Seoul National University College of Medicine, Korea.
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I. Sonoporation

SonoporationÀº ÃÊÀ½ÆÄ¿¡ ÀÇÇØ ¼¼Æ÷¸·ÀÇ Åõ°ú¼ºÀÌ Áõ°¡µÇ´Â Çö»óÀ» ¸»Çϸç, ¼¼Æ÷ ³»·Î °Å´ëºÐÀÚ¸¦ ºñħ½ÀÀûÀ¸·Î À̼ÛÇÒ ¼ö ÀÖ´Â ±â¼ú·Î °¢±¤À» ¹Þ°í ÀÖ´Ù (1). SonoporationÀº ¼¼Æ÷ ÁÖÀ§¿¡ ±âÆ÷°¡ Á¸ÀçÇÒ¶§ ±Þ°ÝÇÏ°Ô Áõ°¡Çϴµ¥ ÀÌ´Â pulsating bubble°ú ¼¼Æ÷ »çÀÌÀÇ fluid dynamic interaction ¶§¹®À̸ç, ±âÆ÷·Î ºÎÅÍ ºÐÃâµÇ´Â shock waves or acoustic transients, extensional flow, shearing flow, ±âÆ÷°¡ ºñ´ëĪÀûÀ¸·Î ÅÍÁú ¶§ ¹æÃâµÇ´Â micro-jetting flow µîÀÌ °ü¿©ÇÏ´Â °ÍÀ¸·Î ¾Ë·ÁÁ® ÀÖ´Ù (2). Jet flow´Â ±âÆ÷°¡ ÆØâ ÈÄ ¾ÐÃàµÉ ¶§ ¹ß»ýÇÏ¸ç ¼Óµµ´Â 5m/s¿¡¼­ 80m/s¿¡ À̸¥´Ù. Single shot ultrasound pulse¿¡ ÀÇÇؼ­µµ ÁÖÀ§ ¾à 1/4ÀÇ ¼¼Æ÷°¡ ÀÌjet flow¿¡ ÀÇÇØ ¼Õ»óÀ» ÀԴ´٠(3). ¼¼Æ÷°¡ ±âÆ÷ ÃÖ´ë¹ÝÁö¸§ÀÇ 0.75¹è ¾È¿¡ ÀÖÀ» ¶§ 75% ÀÌ»óÀÇ È®·ü·Î porationÀÌ ÀϾ´Ù (4). ±×·¯³ª ¾î´À Á¤µµ bubble°ú ¶³¾îÁ® ÀÖÀ» ¶§´Â jet flowÀÇ Á÷Á¢ Ÿ°Ýº¸´Ù´Â jet flow³ª oscillation bubble¿¡ ÀÇÇÑ streaming ¹× shear force°¡ ´õ sonoporation¿¡ Å« ¿ªÇÒÀ» ÇÏ´Â °ÍÀ¸·Î ¾Ë·ÁÁ® ÀÖ´Ù (5, 6). ÀÌ·± sonomechanical effect¿¡ ÀÇÇÑ sonoporation¿Ü¿¡ sonochemical effectÀÇ Á¸À縦 ÁÖÀåÇÏ´Â ±×·ìµµ ÀÖ´Ù. Inertial cavitationÀº microsecond¿Í nanometer scale»ó¿¡¼­ ¸Å¿ì °Ý·ÄÇÑ ¹ÝÀÀÀ̸ç ÀÌ´Â bubble ¼Ó water vaporÀÇ ¿­ºÐÇظ¦ ÀÏÀ¸ÄÑ ¸Å¿ì ³ôÀº ¹ÝÀÀ¼ºÀ» º¸ÀÌ´Â OH radical°ú H atom¸¦ »ý¼ºÇÏ°í ÀÌ radicalµéÀº ÁÖÀ§ ºÐÀÚ¿Í ¹ÝÀÀÇÏ¿© »õ·Î¿î free radicalµéÀ» »ý¼ºÇϰųª DNAÀÇ ¿°±â¼­¿­¿¡ ¿µÇâÀ» ÁØ´Ù. ÀÌ °úÁ¤¿¡¼­ Á» ´õ ¿À·¡ Áö¼ÓµÇ´Â È­Çй°ÁúÀÌ »ý¼ºµÉ ¼ö ÀÖ°í À̵éÀº ÃÊÀ½ÆÄ¿¡ Á÷Á¢ ³ëÃâµÇÁö ¾ÊÀº ¼¼Æ÷¿¡µµ ¹ÙÀÌ¿ÀÈ¿°ú¸¦ ³ªÅ¸³¾ ¼ö ÀÖ´Ù´Â °ÍÀÌ sonochemical effect ÁÖÀåÀÇ ±Ù°£À» ÀÌ·é´Ù. Sonoporation¿¡ ¿µÇâÀ» ³¢Ä¡´Â ÀÎÀÚµéÀº acoustic pressure, pulse frequency, pulse repetition frequency, Pulse duration, insonation time µîÀ̸ç pulse frequency¸¦ Á¦¿ÜÇÑ ³ª¸ÓÁö ¸ðµç ÀÎÀÚµéÀº Å©±â°¡ Ŭ¼ö·Ï cell permeability°¡ Áõ°¡ÇÏ°í viability°¡ °¨¼ÒÇÑ´Ù (7). À¯ÀԵǴ ÀÔÀÚÀÇ size´Â delivery¿¡ Å« ¿µÇâÀ» ÁÖÁö ¾ÊÀ¸¸ç 2 MDa±îÁö À¯ÀÔÀÌ °¡´ÉÇÏ´Ù.

II. Preclinical application

1. Drug delivery (chemotherapeutic agents)
ÃÊÀ½ÆÄ¿Í microbubble(MB)ÀÇ º´ÇÕÀº Ç×¾ÏÁ¦ÀÇ ¼¼Æ÷ ³» À̼ÛÀ» Áõ°¡½ÃÄÑ Ç×¾ÏÁ¦ÀÇ È¿°ú¸¦ ÁõÁø½ÃŲ´Ù. ±× È¿°ú´Â ÃÊÀ½ÆÄ Á¶»ç ÈÄ 60ºÐ ÀÌ»ó Áö¼ÓµÈ´Ù (8). Squamous carcinoma cell¿¡ bleomycinÀ» ÁÖÀÔ ÈÄ MB¸¦ ÀÌ¿ëÇÑ sonoporationÀ» 4ÁÖ ½ÃÇàÇÏ¿´À» ¶§ Á¾¾çÀÌ °ÅÀÇ ¼Ò½ÇµÇ¾ú°í Á¶Á÷ÇÐÀûÀ¸·Î apoptosis°¡ ÁÖ¿´´Ù (9). À̶§ »ç¿ëÇÑ sonicationÀÇ Á¶°ÇÀº 1 MHz, 2.0 W/cm2, duty cycle 50%, 1ºÐ(x2) À̾ú´Ù. Retinoblastoma, primary cutaneous melanoma, lympohoma, prostate cancer, oral cancer, pancreas cancer µî¿¡¼­ º´ÇÕÈ¿°ú°¡ ½ÇÇèÀûÀ¸·Î Áõ¸íÀÌ µÇ°í ÀÖ´Ù (8, 10).
Drug ¿Ü¿¡µµ Protein, synthetic oligonucleotides, small compound µîÀÇ influx¸¦ Áõ°¡½Ãų ¼ö ÀÖ´Ù. ÃÊÀ½ÆÄ¿¡ ÀÇÇÑ transmembrane delivery´Â endocytosis°¡ °ü°èÇÏÁö ¾Ê°í ¾Ë·ÁÁø ´ë·Î pore¿¡ ÀÇÇÑ Á÷Á¢ÀûÀÎ À¯ÀÔÀ¸·Î º¸Àδٴ ÁÖÀåÀÌ ¸¹À¸³ª (11), gene transfectionÀ» Áõ°¡½ÃÅ°±â À§ÇØ polyethylenimine (PEI)¸¦ ÀÌ¿ëÇÑ ¿¬±¸¿¡¼­ ÃÊÀ½ÆÄÁ¶»ç ÈÄ ´ëºÎºÐÀÇ uptake°¡ 3½Ã°£ ¾È¿¡ ÀϾ endocytosis°¡ °ü¿©ÇÒ °¡´É¼ºÀ» Á¦½ÃÇÏ¿´´Ù (12). ÀÌ¿¡ ´ëÇؼ­´Â Á» ´õ ¸¹Àº ¿¬±¸°¡ ÇÊ¿äÇÒ °ÍÀ¸·Î »ý°¢ÀÌ µÈ´Ù (8). MBÀÇ shellÀÇ ¼ººÐ¿¡ ÀÇÇؼ­µµ transfectionÀÇ efficiency°¡ ¿µÇâÀ» ¹Þ´Â µ¥ lipid bubbleÀÌ albumin shellÀ̳ª liposomeº¸´Ù ³´´Ù´Â º¸°í°¡ ÀÖ´Ù (13). Doxorubicin-containing liposomecontaining MBÀ» ÀÌ¿ëÇÑ sonication ½ÇÇè¿¡¼­ Doxliposomeº¸´Ù Dox-loaded MB¿¡¼­ 2¹è ÀÌ»ó melanoma ¼¼Æ÷°¡ »ç¸êµÇ¾ú´Ù (14).

2. Gene delivery
ÃÊÀ½Æĸ¦ ¸Å°³·Î ÇÑ gene transfer´Â viral gene transferÀÇ °¡´ÉÇÑ ´ë¾ÈÀ¸·Î Àû±ØÀûÀ¸·Î ¸ð»öµÇ°í ÀÖ°í ¸¹Àº ÀüÀÓ»ó ¿¬±¸µéÀÌ À̸¦ ÁöÁöÇØ ÁÖ°í ÀÖ´Ù (15). Bacteria ³»¿¡ mutant¸¦ ¸¸µé±â À§ÇØ gene transfectionÀ» ½ÃÅ°´Â µ¥ ÀÖ¾î sonoporationÀº ¸Å¿ì À¯¿ëÇϸç ÇÑ ¿¬±¸¿¡ ÀÇÇϸé electoporationº¸´Ù 2¹è Á¤µµ È¿°úÀûÀ̾ú°í electroporation°ú ´Þ¸® bacteriocidalÇÏÁö ¾Ê¾Ò´Ù. Delivery efficiency´Â acoustic pressure amplitude, duty cycle, ±×¸®°í MBÀÇ ¾ç¿¡ ´Þ·Á ÀÖÀ¸¸ç PRF¿Í´Â Å« ¿µÇâÀ» ¹ÞÁö ¾Ê¾Ò´Ù (16).
Reporter DNA plasmid¸¦ rat eyeÀÇ ciliary muscle¿¡ trasnfectionÇÏ´Â µ¥ US + MB°¡ MB only³ª US only, ¶Ç´Â plasmid only¿¡ ºñÇØ Åë°èÀûÀ¸·Î À¯ÀÇÇÏ°Ô ³ôÀº transfectionÀ» º¸¿´°í lens³ª ciliary muscleÀÇ ¼Õ»óÀÌ ¾ø¾ú´Ù (17). US-mediated antiangiogenic gene therapy¸¦ Ç×¾ÏÄ¡·á³ª ¸é¿ªÄ¡·á¿Í °áÇÕÇÏ¿© Ä¡·áÇÏ¿´À» ¶§ ´õ ³ôÀº È¿°ú¸¦ ¾òÀ» ¼ö ÀÖ¾ú´Ù (18). Electroporation°ú sonoporationÀ» °°ÀÌ ÀÌ¿ëÇÑ °æ¿ì electroporation¸¸ ÀÌ¿ëÇßÀ» ¶§ º¸´Ù 6¹è ÀÌ»ó transfection efficiency°¡ Áõ°¡Çß´Ù (19). Neuronal cell¿¡µµ ¾à 20% transfection efficiency¸¦ º¸ÀÏ Á¤µµ·Î gene transfectionÀÌ Áõ°¡½ÃŲ´Ù (20). MicroRNA transfectionµµ sonoporationÀ» ÀÌ¿ëÇØ Áõ°­½Ãų ¼ö ÀÖ´Ù (21). BBB¸¦ Åë°úÇؼ­ drug ¹× geneÀ» À̼ÛÇÒ ¼ö ÀÖ´Ù (22).

III. Future challenges

ÀÓ»óÀ¸·Î °¡±â À§Çؼ­´Â ´õ ¸¹Àº ½ÇÇè data¿Í ÇÔ²² ÀÓ»óÀûÀ¸·Î Á» ´õ Á¤È®ÇÏ°Ô deep seated tumor¸¦ treatÇÏ°í targeting ÇÒ ¼ö ÀÖ´Â more sophisticated US generating device°¡ ÇÊ¿äÇÏ´Ù. ¶ÇÇÑ ½ÇÁ¦ Á¾¾çÀÌ °¡Áö°í Àִ Ư¼º¿¡ µû¸¥ Ä¡·áÈ¿°úÀÇ Â÷À̸¦ ±Øº¹Çϱâ À§ÇÑ ¿¬±¸°¡ ÇÊ¿äÇѵ¥ Á¾¾çÀÇ heterogeneity¿¡ µû¸¥ À§Ä¡º° sonoporationÀÇ Â÷À̸¦ ¾î¶»°Ô ±Øº¹ÇÒ °ÍÀΰ¡°¡ °ü°ÇÀÌ´Ù. Á¾¾ç³» Ç÷°üÀÌ ¸¹Àº °÷¿¡´Â MBÀÌ Vascular permeability¸¦ Áõ°¡½ÃÄÑ À¯ÃâÀ» Áõ°¡½ÃÅ°Áö¸¸, Ç÷°üÀÌ Àû°Å³ª ±«»çµÈ ºÎºÐ¿¡¼­´Â À¯ÃâÀÌ Á¦ÇѵȴÙ. ÀÌ·± Á¦ÇÑÁ¡Àº nano-sized bubbleÀ̳ª microbubbleÀ» ¸¸µé¾î³»´Â nanoemulsion ¶Ç´Â nanoparticle·Î ±Øº¹ÇÒ ¼ö ÀÖÀ» °ÍÀÌ´Ù
(23-25).

References
1. Deng CX, Sieling F, Pan H, Cui J. Ultrasound-induced cell membrane porosity. Ultrasound Med Biol. 2004;30(4):519-26
2. Ohl CD, Arora M, Ikink R, et al. Sonoporation from jetting cavitation bubbles. Biophys J. 2006;91(11):4285-95
3. Kudo N, Okada K, Yamamoto K. Sonoporation by single-shot pulsed ultrasound with microbubbles adjacent to cells. Biophys J. 2009;96(12):4866-76
4. Gac SL, Zwaan E, van den Berg A, Ohl CD. Sonoporation of suspension cells with a single cavitation bubble in a microfluidic confinement. Lab Chip. 2007;7(12):1666-72
5. Forbes MM, Steinberg RL, O¡¯Brien WD, Jr. Examination of inertial cavitation of Optison in producing sonoporation of chinese hamster ovary cells. Ultrasound Med Biol. 2008;34(12):2009-18
6. Forbes MM, Steinberg RL, O¡¯Brien WD, Jr. Frequency-dependent evaluation of the role of definity in producing sonoporation of Chinese hamster ovary cells. J Ultrasound Med. 2011;30(1):61-9
7. Karshafian R, Bevan PD, Williams R, Samac S, Burns PN. Sonoporation by ultrasound-activated microbubble contrast agents: effect of acoustic exposure parameters on cell membrane permeability and cell viability. Ultrasound Med Biol. 2009;35(5):847-60
8. Lee NG, Berry JL, Lee TC, et al. Sonoporation enhances chemotherapeutic efficacy in retinoblastoma cells in vitro. Invest Ophthalmol Vis Sci. 2011;52(6):3868-73
9. Iwanaga K, Tominaga K, Yamamoto K, et al. Local delivery system of cytotoxic agents to tumors by focused sonoporation. Cancer Gene Ther. 2007;14(4):354-63
10. Lee JY, Choi BI, Ryu JK, et al. Concurrent chemotherapy and pulsed high-intensity focused ultrasound therapy for the treatment of unresectable pancreatic cancer: initial experiences. Korean J Radiol. 2011;12(2):176-86
11. Geers B, Lentacker I, Alonso A, et al. Elucidating the mechanisms behind sonoporation with adeno-associated virusloaded microbubbles. Mol Pharm. 2011;8(6):2244-51
12. Lee JL, Lo CW, Ka SM, Chen A, Chen WS. Prolonging the expression duration of ultrasound-mediated gene transfection using PEI nanoparticles. J Control Release. 2012;160(1):64- 71
13. Kodama T, Aoi A, Watanabe Y, et al. Evaluation of transfection efficiency in skeletal muscle using nano/microbubbles and ultrasound. Ultrasound Med Biol. 2010;36(7):1196-205
14. Lentacker I, Geers B, Demeester J, De Smedt SC, Sanders NN. Design and evaluation of doxorubicin-containing microbubbles for ultrasound-triggered doxorubicin delivery: cytotoxicity and mechanisms involved. Mol Ther. 2010;18(1):101-8
15. Nomikou N, McHale AP. Microbubble-enhanced ultrasoundmediated gene transfer--towards the development of targeted gene therapy for cancer. Int J Hyperthermia. 2012;28(4):300- 10
16. Han YW, Ikegami A, Chung P, Zhang L, Deng CX. Sonoporation is an efficient tool for intracellular fluorescent dextran delivery and one-step double-crossover mutant construction in Fusobacterium nucleatum. Appl Environ Microbiol. 2007;73(11):3677-83
17. Kowalczuk L, Boudinet M, El Sanharawi M, et al. In vivo gene transfer into the ocular ciliary muscle mediated by ultrasound and microbubbles. Ultrasound Med Biol. 2011;37(11):1814-27
18. Liao ZK, Tsai KC, Wang HT, et al. Sonoporation-mediated anti- angiogenic gene transfer into muscle effectively regresses distant orthotopic tumors. Cancer Gene Ther. 2012;19(3):171- 80
19. Escoffre JM, Kaddur K, Rols MP, Bouakaz A. In vitro gene transfer by electrosonoporation. Ultrasound Med Biol. 2010;36(10):1746-55
20. Fischer AJ, Stanke JJ, Omar G, Askwith CC, Burry RW. Ultrasound-mediated gene transfer into neuronal cells. J Biotechnol. 2006;122(4):393-411
21. Chen ZY, Liang K, Qiu RX, Luo LP. Enhancing microRNA transfection to inhibit survivin gene expression and induce apoptosis: could it be mediated by a novel combination of sonoporation and polyethylenimine? Chin Med J (Engl). 2011;124(21):3592-4
22. Sheikov N, McDannold N, Vykhodtseva N, Jolesz F, Hynynen K. Cellular mechanisms of the blood-brain barrier opening induced by ultrasound in presence of microbubbles. Ultrasound Med Biol. 2004;30(7):979-89
23. Horie S, Watanabe Y, Ono M, Mori S, Kodama T. Evaluation of antitumor effects following tumor necrosis factor-alpha gene delivery using nanobubbles and ultrasound. Cancer Sci. 2011;102(11):2082-9
24. Rapoport NY, Kennedy AM, Shea JE, Scaife CL, Nam KH. Controlled and targeted tumor chemotherapy by ultrasoundactivated nanoemulsions/microbubbles. J Control Release. 2009;138(3):268-76
25. Kang E, Min HS, Lee J, et al. Nanobubbles from gas-generating polymeric nanoparticles: ultrasound imaging of living subjects. Angew Chem Int Ed Engl. 2010;49(3):524-8

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