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4. A solution to the biophysical fractionation of extracellular vesicles: Acoustic Nanoscale Separation via Wave-pillar Excitation Resonance (ANSWER)

Author

Zhang, Jinxin, Chen, Chuyi, Becker, Ryan, Rufo, Joseph, Yang, Shujie, Mai, John, Zhang, Peiran, Gu, Yuyang, Wang, Zeyu, Ma, Zhehan, Xia, Jianping, Hao, Nanjing, Tian, Zhenhua, Wong, David TW, Sadovsky, Yoel, Lee, Luke P, and Huang, Tony Jun

Subjects
Physical Sciences, Classical Physics
Abstract

High-precision isolation of small extracellular vesicles (sEVs) from biofluids is essential toward developing next-generation liquid biopsies and regenerative therapies. However, current methods of sEV separation require specialized equipment and time-consuming protocols and have difficulties producing highly pure subpopulations of sEVs. Here, we present Acoustic Nanoscale Separation via Wave-pillar Excitation Resonance (ANSWER), which allows single-step, rapid (96% small exosomes, >80% exomeres) fractionation of sEV subpopulations from biofluids without the need for any sample preprocessing. Particles are iteratively deflected in a size-selective manner via an excitation resonance. This previously unidentified phenomenon generates patterns of virtual, tunable, pillar-like acoustic field in a fluid using surface acoustic waves. Highly precise sEV fractionation without the need for sample preprocessing or complex nanofabrication methods has been demonstrated using ANSWER, showing potential as a powerful tool that will enable more in-depth studies into the complexity, heterogeneity, and functionality of sEV subpopulations.

Published
2022

5. Phase 2 of extracellular RNA communication consortium charts next-generation approaches for extracellular RNA research

Author

Mateescu, Bogdan, Jones, Jennifer C, Alexander, Roger P, Alsop, Eric, An, Ji Yeong, Asghari, Mohammad, Boomgarden, Alex, Bouchareychas, Laura, Cayota, Alfonso, Chang, Hsueh-Chia, Charest, Al, Chiu, Daniel T, Coffey, Robert J, Das, Saumya, De Hoff, Peter, deMello, Andrew, D’Souza-Schorey, Crislyn, Elashoff, David, Eliato, Kiarash R, Franklin, Jeffrey L, Galas, David J, Gerstein, Mark B, Ghiran, Ionita H, Go, David B, Gould, Stephen, Grogan, Tristan R, Higginbotham, James N, Hladik, Florian, Huang, Tony Jun, Huo, Xiaoye, Hutchins, Elizabeth, Jeppesen, Dennis K, Jovanovic-Talisman, Tijana, Kim, Betty YS, Kim, Sung, Kim, Kyoung-Mee, Kim, Yong, Kitchen, Robert R, Knouse, Vaughan, LaPlante, Emily L, Lebrilla, Carlito B, Lee, L James, Lennon, Kathleen M, Li, Guoping, Li, Feng, Li, Tieyi, Liu, Tao, Liu, Zirui, Maddox, Adam L, McCarthy, Kyle, Meechoovet, Bessie, Maniya, Nalin, Meng, Yingchao, Milosavljevic, Aleksandar, Min, Byoung-Hoon, Morey, Amber, Ng, Martin, Nolan, John, De Oliveira, Getulio P, Paulaitis, Michael E, Phu, Tuan Anh, Raffai, Robert L, Reátegui, Eduardo, Roth, Matthew E, Routenberg, David A, Rozowsky, Joel, Rufo, Joseph, Senapati, Satyajyoti, Shachar, Sigal, Sharma, Himani, Sood, Anil K, Stavrakis, Stavros, Stürchler, Alessandra, Tewari, Muneesh, Tosar, Juan P, Tucker-Schwartz, Alexander K, Turchinovich, Andrey, Valkov, Nedyalka, Van Keuren-Jensen, Kendall, Vickers, Kasey C, Vojtech, Lucia, Vreeland, Wyatt N, Wang, Ceming, Wang, Kai, Wang, ZeYu, Welsh, Joshua A, Witwer, Kenneth W, Wong, David TW, Xia, Jianping, Xie, Ya-Hong, Yang, Kaichun, Zaborowski, Mikołaj P, Zhang, Chenguang, Zhang, Qin, Zivkovic, Angela M, and Laurent, Louise C

Subjects
Biological Sciences, Biomedical and Clinical Sciences, Genetics, Biochemistry, Biological sciences, Cell biology, Molecular biology
Abstract

The extracellular RNA communication consortium (ERCC) is an NIH-funded program aiming to promote the development of new technologies, resources, and knowledge about exRNAs and their carriers. After Phase 1 (2013-2018), Phase 2 of the program (ERCC2, 2019-2023) aims to fill critical gaps in knowledge and technology to enable rigorous and reproducible methods for separation and characterization of both bulk populations of exRNA carriers and single EVs. ERCC2 investigators are also developing new bioinformatic pipelines to promote data integration through the exRNA atlas database. ERCC2 has established several Working Groups (Resource Sharing, Reagent Development, Data Analysis and Coordination, Technology Development, nomenclature, and Scientific Outreach) to promote collaboration between ERCC2 members and the broader scientific community. We expect that ERCC2's current and future achievements will significantly improve our understanding of exRNA biology and the development of accurate and efficient exRNA-based diagnostic, prognostic, and theranostic biomarker assays.

Published
2022

6. An acoustofluidic picoinjector

Author

Naquin, Ty, Jain, Shivam, Zhang, Jinxin, Xu, Xianchen, Yao, Gary, Naquin, Chloe M., Yang, Shujie, Xia, Jianping, Wang, Janna, Jimenez, Sebastian, and Huang, Tony Jun

Published
2024
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8. Acoustofluidic sonoporation for gene delivery to human hematopoietic stem and progenitor cells

Author

Belling, Jason N, Heidenreich, Liv K, Tian, Zhenhua, Mendoza, Alexandra M, Chiou, Tzu-Ting, Gong, Yao, Chen, Natalie Y, Young, Thomas D, Wattanatorn, Natcha, Park, Jae Hyeon, Scarabelli, Leonardo, Chiang, Naihao, Takahashi, Jack, Young, Stephen G, Stieg, Adam Z, De Oliveira, Satiro, Huang, Tony Jun, Weiss, Paul S, and Jonas, Steven J

Subjects
Medical Biotechnology, Biomedical and Clinical Sciences, Stem Cell Research, Biotechnology, Bioengineering, Genetics, Gene Therapy, Stem Cell Research - Nonembryonic - Human, Development of treatments and therapeutic interventions, 5.2 Cellular and gene therapies, Generic health relevance, Cell Survival, Cytoplasm, Gene Expression, Gene Transfer Techniques, Genetic Therapy, Green Fluorescent Proteins, Hematopoietic Stem Cell Transplantation, Hematopoietic System, Humans, Jurkat Cells, Plasmids, Sound, Stem Cells, acoustofluidics, hematopoietic stem cells, intracellular delivery, gene therapy
Abstract

Advances in gene editing are leading to new medical interventions where patients' own cells are used for stem cell therapies and immunotherapies. One of the key limitations to translating these treatments to the clinic is the need for scalable technologies for engineering cells efficiently and safely. Toward this goal, microfluidic strategies to induce membrane pores and permeability have emerged as promising techniques to deliver biomolecular cargo into cells. As these technologies continue to mature, there is a need to achieve efficient, safe, nontoxic, fast, and economical processing of clinically relevant cell types. We demonstrate an acoustofluidic sonoporation method to deliver plasmids to immortalized and primary human cell types, based on pore formation and permeabilization of cell membranes with acoustic waves. This acoustofluidic-mediated approach achieves fast and efficient intracellular delivery of an enhanced green fluorescent protein-expressing plasmid to cells at a scalable throughput of 200,000 cells/min in a single channel. Analyses of intracellular delivery and nuclear membrane rupture revealed mechanisms underlying acoustofluidic delivery and successful gene expression. Our studies show that acoustofluidic technologies are promising platforms for gene delivery and a useful tool for investigating membrane repair.

Published
2020

9. Acoustofluidic Salivary Exosome Isolation A Liquid Biopsy Compatible Approach for Human Papillomavirus–Associated Oropharyngeal Cancer Detection

Author

Wang, Zeyu, Li, Feng, Rufo, Joseph, Chen, Chuyi, Yang, Shujie, Li, Liang, Zhang, Jinxin, Cheng, Jordan, Kim, Yong, Wu, Mengxi, Abemayor, Elliot, Tu, Michael, Chia, David, Spruce, Rachel, Batis, Nikolaos, Mehanna, Hisham, Wong, David TW, and Huang, Tony Jun

Subjects
Biomedical and Clinical Sciences, Oncology and Carcinogenesis, Infectious Diseases, Clinical Research, Dental/Oral and Craniofacial Disease, Prevention, Biotechnology, Sexually Transmitted Infections, Cervical Cancer, Digestive Diseases, Cancer, Base Sequence, Biomarkers, Tumor, DNA, Viral, Exosomes, Human papillomavirus 16, Humans, Liquid Biopsy, MicroRNAs, Microfluidics, Oropharyngeal Neoplasms, Papillomavirus Infections, Polymerase Chain Reaction, Prognosis, Saliva, Viscosity, Medical Microbiology, Pathology, Clinical sciences, Oncology and carcinogenesis
Abstract

Previous efforts to evaluate the detection of human papilloma viral (HPV) DNA in whole saliva as a diagnostic measure for HPV-associated oropharyngeal cancer (HPV-OPC) have not shown sufficient clinical performance. We hypothesize that salivary exosomes are packaged with HPV-associated biomarkers, and efficient enrichment of salivary exosomes through isolation can enhance diagnostic and prognostic performance for HPV-OPC. In this study, an acoustofluidic (the fusion of acoustics and microfluidics) platform was developed to perform size-based isolation of salivary exosomes. These data showed that this platform is capable of consistently isolating exosomes from saliva samples, regardless of viscosity variation and collection method. Compared with the current gold standard, differential centrifugation, droplet digital RT-PCR analysis showed that the average yield of salivary exosomal small RNA from the acoustofluidic platform is 15 times higher. With this high-yield exosome isolation platform, we show that HPV16 DNA could be detected in isolated exosomes from the saliva of HPV-associated OPC patients at 80% concordance with tissues/biopsies positive for HPV16. Overall, these data demonstrated that the acoustofluidic platform can achieve high-purity and high-yield salivary exosome isolation for downstream salivary exosome-based liquid biopsy applications. Additionally, HPV16 DNA sequences in HPV-OPC patients are packaged in salivary exosomes and their isolation will enhance the detection of HPV16 DNA.

Published
2020

13. Acoustothermal transfection for cell therapy

Author

Liu, Xiufang, primary, Rong, Ning, additional, Tian, Zhenhua, additional, Rich, Joseph, additional, Niu, Lili, additional, Li, Pengqi, additional, Huang, Laixin, additional, Dong, Yankai, additional, Zhou, Wei, additional, Zhang, Pengfei, additional, Chen, Yizhao, additional, Wang, Congzhi, additional, Meng, Long, additional, Huang, Tony Jun, additional, and Zheng, Hairong, additional

Published
2024
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23. List of contributors

Author

Bautista, Cynthia, primary, Cao, Lei, additional, Chen, Hui, additional, Du, Ke, additional, Fillet, Marianne, additional, Hass, Kenneth, additional, He, Yong, additional, Hu, Huan, additional, Huang, Tony Jun, additional, Isaac, K.M., additional, Jiang, Dongyue, additional, Li, Nan, additional, Li, Paul C.H., additional, Li, XiuJun, additional, Li, Zedong, additional, Lin, Jin-Ming, additional, Ma, Lei, additional, Nie, Jing, additional, Nix, Cindy, additional, Oberc, Christopher A., additional, Ozcelik, Adem, additional, Qin, Peiwu, additional, Rich, Joseph, additional, Sun, Jiashu, additional, Sun, Yue, additional, Tavakoli, Hamed, additional, Wang, C., additional, Wang, Chuhui, additional, Xu, Feng, additional, Xu, Yan, additional, Xu, Zhiheng, additional, Yang, Bin, additional, Yang, Jinbin, additional, Yang, Zhejun, additional, Yao, Bo, additional, Yeo, Leslie Y., additional, You, Minli, additional, Yuan, F., additional, Yuan, Xi, additional, Zhang, J., additional, Zhang, Lu, additional, Zhang, Peng, additional, Zhang, Weifei, additional, and Zhou, Wan, additional

Published
2022
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25. High-yield and rapid isolation of extracellular vesicles by flocculation via orbital acoustic trapping: FLOAT

Author

Rufo, Joseph, primary, Zhang, Peiran, additional, Wang, Zeyu, additional, Gu, Yuyang, additional, Yang, Kaichun, additional, Rich, Joseph, additional, Chen, Chuyi, additional, Zhong, Ruoyu, additional, Jin, Ke, additional, He, Ye, additional, Xia, Jianping, additional, Li, Ke, additional, Wu, Jiarong, additional, Ouyang, Yingshi, additional, Sadovsky, Yoel, additional, Lee, Luke P., additional, and Huang, Tony Jun, additional

Published
2024
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28. Acoustic Streaming: An Arbitrary Lagrangian-Eulerian Perspective

Author

Nama, Nitesh, Huang, Tony Jun, and Costanzo, Francesco

Subjects
Physics - Fluid Dynamics
Abstract

We analyze acoustic streaming flows using an ALE perspective. The formulation stems from an explicit separation of time-scales resulting in two subproblems: a first-order problem, formulated in terms of the fluid displacement at the fast scale, and a second-order problem formulated in terms of the Lagrangian flow velocity at the slow time scale. Following a rigorous time-averaging procedure, the second-order problem is shown to be intrinsically steady, and with exact boundary conditions at the oscillating walls. Also, as the second-order problem is solved directly for the Lagrangian velocity, the formulation does not need to employ the notion of Stokes drift, or any associated post-processing, thus facilitating a direct comparison with experiments. Because the first-order problem is formulated in terms of the displacement field, our formulation is directly applicable to more complex fluid-structure interaction problems in microacosutofluidic devices. After the formulation's exposition, we present numerical results that illustrate the advantages of the formulation with respect to current approaches.

Published
2016
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30. Cellular immunity analysis by a modular acoustofluidic platform: CIAMAP

Author

Zhong, Ruoyu, primary, Sullivan, Matthew, additional, Upreti, Neil, additional, Chen, Roy, additional, De Ganzó, Agustin, additional, Yang, Kaichun, additional, Yang, Shujie, additional, Jin, Ke, additional, He, Ye, additional, Li, Ke, additional, Xia, Jianping, additional, Ma, Zhiteng, additional, Lee, Luke P., additional, Konry, Tania, additional, and Huang, Tony Jun, additional

Published
2023
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31. Investigation of acoustic streaming patterns around oscillating sharp edges

Author

Nama, Nitesh, Huang, Po-Hsun, Huang, Tony Jun, and Costanzo, Francesco

Subjects
Physics - Fluid Dynamics
Abstract

Oscillating sharp edges have been employed to achieve rapid and homogeneous mixing in microchannels using acoustic streaming. Here we use a perturbation approach to study the flow around oscillating sharp edges in a microchannel. This work extends prior experimental studies to numerically characterize the effect of various parameters on the acoustically induced flow. Our numerical results match well with the experimental results. We investigated multiple device parameters such as the tip angle, oscillation amplitude, and channel dimensions. Our results indicate that, due to the inherent nonlinearity of acoustic streaming, the channel dimensions could significantly impact the flow patterns and device performance., Comment: 15 pages, 11 figures, submitted for publication in " Lab on a Chip" on 12 February 2014, micro-acousto-fluidics, streaming flows, mixing

Published
2014

42. Contributors to the emerging investigators issue.

Author

Basu, Amar, Cheung, Karen C, Eddington, David T, Günther, Axel, Hansen, Carl, Huang, Tony Jun, Juncker, David, Kaji, Hirokazu, Khademhosseini, Ali, Khan, Saif A, Klapperich, Catherine, Love, J Christopher, Munson, Matthew, Murthy, Shashi, Ozcan, Aydogan, Ozinsky, Adrian, Spotts, James M, Squires, Todd, Takeuchi, Shoji, Wang, Wei, and Williams, Justin

Subjects
Analytical Chemistry, Chemical Sciences, Engineering
Published
2010

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