Pressure, gravity, waves, temperature, light, and electric and magnetic fields make up an omnipresent physical presence since the beginning of time. It should be no surprise, therefore, that the capacity of biologic systems to adapt to physical signals is a common attribute for essentially all life, including bacteria, yeast, plant and animal cells, and that the cellular machinery responsible for sensing and responding to mechanical signals might be even more evolved than those processes that are regulated by complex macromolecules.
All cells respond to unique mechanical stimuli. Lahara Bio uses cell-specific mechanical signals to expand cells quicker. High frequency, low intensity signals can influence biological outcomes.
Laharia bio meets bioprocessing requirements and improves cell-specific biomanufacturing,
Our solution is non-invasive, autonomous, self-adjusting, and ready to incorporate into biomanufacturing.
To facilitate and semi-automate the testing of various combinations of LIV signal parameters, a closed-loop feedback-controlled LIV system has been designed for use inside the cell culture incubator. The generic elements of the LIV system include an electromagnetic actuator base supported by a surrounding glider-spring system to deliver smooth sinusoidal signals via vertical oscillations, with current controlling displacement established via closed-loop error feedback. As designed, all major control components are protected from the high humidity environment of the incubator, and processor controls allow for autonomous 24/7 regulation and monitoring of the mechanical delivery system. As scale-up protocols develop, means of delivering the LIV signal to multiple cell products, simultaneously, “simply” by conveying a controlled vibration to the system. There is a range of issued and pending patents related both to the delivery of LIV to in vitro and in vivo systems, as well as to the signal itself, including refractory periods.
To show the importance of signal optimization, we have results from signals tailored to CHO adherent cells. On the left, frequency optimization of CHO adherent cells is shown. When CHO cells were stimulated with frequency A1, we saw between 70-80% more cells in 48 hours when compared to control. On the right, magnitude is the optimized parameter. Subjected to magnitude B2, there were between 20-30% more cells over 48 hours than in the control group. These results demonstrated that cells respond to specific mechanical signal parameters. For further application we have applied the Lahara LIV signal to Pan T cells. At the optimal frequency and daily dosage, results showed between 20-30 and 30-40% more cells respectively. This demonstrates our signal can be modified to increase proliferation in any cell type, including T-cells used in CAR-T therapies.
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On the left, a combination of the optimized frequency A1 and optimized magnitude B2 show an increase of between 80-100% more cells over 48 hours in CHO adherent cells. On the right, A1 and B2 were applied CHO suspension cells. The same signal applied to CHO suspension cells suppressed cell proliferation by 10%. Changing the signal parameters to CHO suspension cells with combination of A3 and B4 Showed between 150-250% more cells over a 48 hour period. Thus proving that Lahara technology can augment cell proliferation rates of multiple cell types by specifying parameters to each cell.
Working at an industry scale in T75 flasks with Pan T cells expanded for 5 days. Cells were subjected to an optimized Lahara signal for the traditional 48 hour period, and then continued to receive the same treatment for an additional 72 hours. To the 48 hour mark we saw 30-45% more cells in the Lahara treated group as compared to control. Continuing past the 48 hour mark up to 120 hours the Lahara signal is considered to have a significant effect. Treated cells reached final numbers 30-40% higher than control cells.
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Lahara Biotechnology’s work is targeted towards understanding how mechanical factors influence mediate morphologic, cellular and molecular responses through the control of cell differentiation and proliferation. Lahara Bio, a start-up focused on translating Low Intensity Vibration (LIV) technology to in vitro applications, focusing on T-cell expansion for autologous cell therapies. Four publications which provide a broad scientific perspective of the scientific foundations of Lahara on mechanical/electrical/ultrasound factors and bone formation/regeneration, as well as our efforts to translate this to the clinic, include:
1. Bone Anabolism by LMMS/LIV: Rubin, C., Turner, S. Bain, S., Mallinckrodt, C. & McLeod, K. (2001) Anabolism: Low mechanical signals strengthen long bones. Nature 412:603-604.2.
2. Use of Physical Signals as a Non-Drug Intervention for Osteoporosis: Rubin, C.T., Recker, R., Raab, D.,Ryaby, J., McCabe, J. & McLeod, K.J. (2004) Prevention of Post-Menopausal Bone Loss by a Low Magnitude, High Frequency Mechanical Stimuli; A Clinical Trial Assessing Compliance, Efficacy And Safety. J. Bone & Mineral Research. 19:343-3513.
3. Ultrasound as a means of Accelerating Fracture Healing: Rubin, C., Bolander, M., Ryaby, J. & Hadjiargyrou, M. (2001) The use of low intensity ultrasound to accelerate the healing of fractures. J.Bone & Jt. Surg. 83:259-70.4.
4. Identifying Electric Field Parameters that Stimulate Bone Formation: Rubin, C.T., McLeod, K.J., and Lanyon, L.E.(1989) Prevention of osteoporosis by pulsed electro‑magnetic fields. J. Bone and Joint Surgery, 71A(3):411‑418 1.
1. Identifying specific mechanical parameters that drive adaptation in bone: We focus on the use of in vivo models of bone adaptation to identify specific mechanical signals which drive the ‘form follows function’ paradigm in the skeleton. While disuse in these models would result in a marked loss of bone, externally applied loading regimens, physiological in strain magnitude, could lead to significant increases in cross-sectional bone area, as dependent on changes in the magnitude and distribution generated within the bone tissue. The loading regimen must be dynamic (time-varying) in nature; static loads do not influence bone morphology. Perhaps most importantly, this work indicates the full osteogenic potential of a large amplitude (>2000 micro strain) regimen is realized following only an extremely short (< 1 min) exposure to this stimulus, suggesting that brief exposure to a specific signal can set off subsequent adaptive events.
a. Rubin, C.T. & Lanyon, L.E. (1984) Regulation of bone formation by applied dynamic loads. J. Bone and Joint Surgery 66A:397‑402. PMID:6699056
b. Pagnotti, G., Styner, M., Uzer, G., Patel, V. Ness, K., Guise, T., Rubin, J& Rubin, C.T. (2019) Combating osteoporosis and obesity with exercise: Harnessing cell mechanosensitivity. Nature Reviews Endocrinology. 15:339-355 PMID: 30814687
c. Fritton, S.P., McLeod, K.J., Rubin, C.T. (2000) Quantifying the strain history of bone: spatial uniformity and self-similarity of low magnitude strains. J. Biomech. 33:317-326. PMID:10673115
d. Ozcivici, E., Luu, Y-K, Adler, B., Qin, Y-X,Rubin, J., Judex, S & Rubin, C.T. (2010) Mechanical Signals as Anabolic Agents in Bone. Nature Reviews Rheumatology. 6:50-59 PMCID: PMC3743048
2. Low magnitude mechanical signals are anabolic to the musculoskeletal system:
Moving away from peak strain magnitudes, this work demonstrated that extremely small magnitude mechanical signals(<10 microstrain), if introduced at a relatively high frequency (>10Hz),were strongly anabolic to the skeleton. This work pointed towards the spectral content of muscle contraction, rather than peak impacts, as important signals in the achieving and retaining bone quality and quantity, even under conditions challenged by disuse or disease.
a. Rubin, C.T., Xu, G. & Judex, S. (2001) The anabolic activity of bone tissue, suppressed by disuse is normalized by brief exposure to extremely low magnitude mechanical stimuli. TheFASEB Journal. 15: 2225-2229 PMID:11641249
b. Goodship, A., Lawes, T. & Rubin, C. (2009) Low magnitude high frequency mechanical signals accelerate and augment endochondral bone repair. J.Orth. Res. 27:922-930 PMCID: PMC2929925
c. Mettlach, G., Polo-Parada, L., Peca, L., Rubin, C.T., Plattner, F. & Bibb, J. (2014) Enhancement of Muscle Dynamics and StrengthBehavior Using Low Magnitude Mechanical Signals. J. Biomechanics 47:162-7 PMCID: PMC3881264
d. Pagnotti, G., Chan, M.E., Adler, B., Rubin, J., Shroyer, K. & Rubin, C.T. (2016) Low Intensity Mechanical Signals Mitigate Tumor Progression andProtect Bone Quantity and Quality in a Murine Model of Myeloma. Bone 90:69-79 PMCID: PMC49708893.
3. Treatment of musculoskeletal disorders with non-invasive mechanical stimulation: Translating the musculoskeletal system’s sensitivity to mechanical signals to the human, a series of double-blind, prospective clinical trials were performed to determine if low intensity vibration could serve as a non-drug means of stimulating growth of bone and muscle in those with poor bone quality.
a. Ward, K., Alsop, C., Brown, S., Caulton, J., Rubin, C., Adams, J. & Mughal, M. (2004) Low magnitude mechanical loading is osteogenic in children with disabling conditions. J. Bone& Mineral Research: 19:360-369 PMID:15040823
b. Gilsanz, V., Wren, T., Sanchez, M., Dorey, F., Judex, S. & Rubin,C.T. (2006) Low level, high frequency mechanical signals enhance musculoskeletal development of young women with low bone density. J.Bone & Mineral Research 21:1464-1474 PMID: 16939405
c. Kiel, D.P., Hannan, M.T., Barton, B., Bouxsein, M.L., Sisson, E., Lang, T., Allaire, B., Dewkett, D., Carroll, D.,Magaziner, J., Shane, E., Leary, E., Zimmerman, S. & Rubin, C.T.,(2015). Low Magnitude MechanicalStimulation to Improve Bone Density in Persons of Advanced Age: A Randomized, Placebo-Controlled Trial. J. Bone& Mineral Research 30:1319-1328 PMCID: PMC4834704
d. Leonard, M., Shults, J., Long, J., Baldassanao, R., Brown, K., Hommel,K., Zemel, B., Mahboubi, S., Whitehead, K., Herskovitz, R., Lee, D. &Rubin, C.T. (2016). Effect of Low Magnitude Mechanical Stimulation on BoneDensity and Structure in Pediatric Crohn Disease: A Randomized Placebo-Controlled Trial. J.Bone & Mineral Research; 31:1177-1188 PMCID:PMC4891301
4. Mechanical biasing of stem cell differentiation to suppress adipogenesis: Honing in on the sensitivity of the musculoskeletal system to these low level mechanical signals, we have now demonstrated that these stimuli markedly bias mesenchymal stem cell differentiation towards osteoblastogenesis (bone formation), and away from adipogenesis (fat formation). Mechanosensitivity of the marrow cell population extends to hematopoietic stem cells, with our work now investigating the influence of these signals as a non-drug means of controlling obesity and T2 diabetes.
a. Luu, Y.K., Capilla, E., Pessin, J., Rosen, C., Gilsanz, V., Judex, S.& Rubin, C. (2009) Mechanicalsimulation of mesenchymal stem cell proliferation and differentiation promotesosteogenesis while preventing dietary-induced obesity. J. Bone & Min. Res. 24:50-61 PMCID: PMC2689082
b. Chan, M.E., Adler, B.J., Green, D.E. &Rubin, C.T. (2012) Bone Structure and B-Cell Populations, Crippled by Obesity, arePartially Rescued by Brief Daily Exposure to Low Magnitude Mechanical Signals J. FASEB 26:4855-63 PMCID: PMC3509057
c. Adler, B., Kaushansky, K. & Rubin, C.T., (2014) Obesity DrivenDisruption of Hematopoiesis and the Bone Marrow Niche. NatureReviews Endocrinology 10:737-748 PMID: 25311396
d. Frechette, D.M., Krishnamoorthy, D., Adler, B., Chan, M.E. & Rubin,C.T. (2015 Diminished satellite cells and elevated adipogenic gene expression in muscle as caused by ovariectomy are averted by low-magnitude mechanical signals. J. Applied Physiology 119:27-36 PMCID: PMC4491530
5. Identifying molecular and physical pathways which foster mechano transduction: This work, done in a long-term collaboration with Janet Rubin, is targeted towards determining key signal transduction pathways, as well as features of cell morphology, that allow the cell to perceive and respond to mechanical signals. By focusing on the molecular mechanisms responsible for mechano sensitivity, we are working towards a better understanding by which extremely low level signals can influence phenotype.
a. Sen, B., Styner, M., Xie, Z., Case, N., Rubin, C.T. & Rubin, J.(2010) Mechanical loading regulates NFATC1 and & b-catenin signaling through a GSK3 & b-control node. J. Biol. Chem 284:34607-34617. PMCID: PMC2787323
b. Sen, B., Guilluy, C., Xie, Z., Case, N., Syner, M., Thomas, J., Oguz,I., Rubin, C.T., Burridge, K. & Rubin, J. (2012) Mechanically induced focal adhesion assembly amplifies anti-adipogenic pathways in mesenchymal stemcells. Stem Cells 29:1829-36 PMCID: PMC3588570
c. Wallace, I., Pagnotti, G., Rubin-Sigler, J., Naeher, M., Judex, S.,Rubin, C.T. & Demes, B. (2015) FocalEnhancement of the Skeleton to Exercise Correlates to Mesenchymal Stem CellResponsivity Rather than Peak Forces. J. Exp. Biology. 218:3002-9 PMCID: PMC4631774
d. Uzer, G., Thompson, W.R., Sen, B., Xie, Z., Yen, S., Miller, S., Bas,G., Styner, M, Rubin, C.T., Judex, S., Burridge, K & Rubin, J. (2015) Cell mechanosensitivity to extremely low magnitude signals is enabled by a LINCed nucleus. StemCells 33:2063-2076 PMCID: PMC4458857
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Lahara Bio has a world class team with decades of experience in science and technology.
Clint is a SUNY Distinguished Professor of Biomedical Engineering, and Director of the Center for Biotechnology at Stony Brook University in Stony Brook, New York. Rubin’s research is targeted towards understanding the cellular mechanisms responsible for the growth, healing, and homeostasis of bone, and how mechanical stimuli mediate these responses through the control of mesenchymal and hematopoietic stem cell differentiation and proliferation, to establish non-drug treatment strategies for osteoporosis, obesity and diabetes. Dr. Rubin holds ~30 patents in the area of wound repair, stem cell regulation, and treatment of metabolic disease, and is a founder of Exogen, Juvent, Marodyne Medical, and Lahara Bio, which use physical signals to regulate biologic processes. He has published over 300 articles, has been cited ~34,000 times, with an H-index of 94. He is a fellow of AAAS, NAI, ASBMR, BMES and AIMBE. Dr. Rubin received his AB degree from Harvard, and his PhD from Bristol University, U.K.
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Michael R. Bielski, J.D., M.S. is a serial entrepreneur and C-level executive in the medical technology and biotechnology fields. Michael has brought technologies from early conception, development and patenting through manufacturing, the generation of licensing revenues, clinical use and sales. Michael is an experienced angel and venture capital fundraiser whose companies have raised over $75M in financing. Michael is an expert across all areas of high-technology commercialization including out-licensing and start-up company formation and operations.
Michael received a B.S. in Biology from Stony Brook University, an M.S. in Neuroscience from Syracuse University, and a J.D. from Syracuse University College of Law, where he was a research associate at the Technology Commercialization Research Center (TCRC).
Ete is the Principal Investigator of the Osteoimmunology lab at Stony Brook University, NY. She received her Ph.D. training in Biomedical Engineering from Columbia University, NY. Her research focuses on mechano biology, which is the study of how mechanical signals influences cellular activities and functions. In addition to understanding the underlying working mechanisms, she has a vast interest in optimizing mechanical signals for various important clinical and biotechnology applications including augmenting biomanufacturing speed and yields (e.g.,therapeutic protein production, autologous cell therapy).
Sishir is a research assistant and graduate student in Dr. Mei Lin Chan’s osteoimmunology lab in the Department of Biomedical Engineering at Stony Brook University located in Stony Brook, NY. His research is targeted towards both understanding the mechanisms behind the mitogenic and proliferative effects of low intensity mechanical vibration and the clinical/industrial translation and optimization of autologous cell-based immunotherapies. His role at Lahara Bio consists of providing key scientific input and conducting research on the use of non-drug, mechanical signals for high-throughput biomanufacturing. After graduating from his acceleratedB.E./M.S. program at Stony Brook University, he aims to pursue an MD/PhD dual degree program with hopes of applying his engineering skills to solving problems in the biomedical field, through clinical translational science.
Lia is a research assistant in Dr. Mei Lin Chan’s osteoimmunology lab, located in the Department of Biomedical Engineering at Stony Brook University, in Stony Brook N.Y. She is an undergraduate student currently pursuing an accelerated B.E./M.S. degree. Her research under Dr. Chan seeks to elucidate the underlying mechanisms within cells that result in the augmentation of proliferative behavior when subjected to low-intensity mechanical signals. Her role at Lahara Bio is the design and execution of experimental research on their mechanical signals, to be implemented into biomanufacturing processes. After graduation, Lia hopes to pursue a Ph.D. with goals of applying her knowledge of the impact of mechanical signals to neural stem cells.
Chris is a research assistant and 3rd year MD/PhD student in the lab of Dr. Clinton Rubin in the Department of Biomedical Engineering at Stony Brook University located in Stony Brook, NY. His research is directed towards improving the efficacy of CAR-T therapy by elucidating the mechanisms behind low-intensity vibration's effect on immune cells and understanding how mechanical signals can drive cell phenotype and effector function. His role at Lahara Bio consists of providing key scientific input and conducting research on the use of non-drug, mechanical signals, for high-throughput biomanufacturing of autologous cell therapies. After graduating from his MD/PhD dual degree program he hopes to continue to pursue translational projects with the goal of improving immunotherapies for cancer.
Dr. Rubin practices Endocrinology in the Meadowmont Endo/Diabetes Center with a specialty in Osteoporosis and metabolic bone disease, and is responsible for bone density testing there. Dr. Rubin investigates the controls over bone remodeling, in particular exercise and mechanical force effects on the cell cytoskeleton. At the University Dr Rubin continues her decades long research into bone remodeling. The laboratory is currently investigating how bone marrow stem cells are directed to become osteoblasts or adipocytes, with a particular interest in how exercise regulates this divergent differentiation through the cellular cytoskeleton. Her research is funded by the NIH. Dr. Rubin’s lab recently published a case detailing an important side effect of iron administration, FGF23 based rickets, which was written with a medical intern at UNC.Dr. Rubin is also active in DOM leadership as the Vice Chair for Research. Her interests are in junior faculty research development, and promotion of research throughout the Department.
MD: Brown University, 1980; Intern and Resident: Northwestern University Medical Center, 1980-1983; Endocrinology and Metabolism Fellowship: University of California, San Diego, 1980-1983; Instructor Medicine: UCSD, 1983-1986; Faculty and Professor of Medicine with tenure: Emory University, 1986-2006; Physician: VAMC Atlanta, 1986-2006; Professor of Medicine with tenure: University of North Carolina, 2006-present
Gunes is an Assistant Professor in the Department of Mechanical and Biomedical Engineering at Boise State University. Uzer is the director of the Mechanical Adaptations Laboratory (MAL) leading a multidisciplinary research program. Studies in Mechanical Adaptations Laboratory are directed towards understanding the physiological mechanical environment of cells by establishing experimental and computational models as well as developing novel bioreactor and tissue analog systems to re-create complex loading environments in the bone marrow. These models and analog systems are utilized to understand how changes in tissue mechanical environment in relation to exercise, injury, aging, disuse and space travel regulate structural adaptations and signaling of stem cells. Research at MAL is actively funded by NIH, NSF and NASA for technology development and translational science projects.
