Abstract: Tissue Ovarian Cancer
Case Report CRLR-2-100012
Vibratess to participate in PITCONN
The biggest achievements in the past 12 months?
a) Experimental demonstration of uniqueness of spectroscopic signatures produced by diverse microRNA molecules using fast spectroscopy analysis of small samples from biopsies of patients with cancer and normal epithelia control tissue. This uniqueness makes it possible to detect cancer at its early stage, to identify and discriminate different cancers (ovarian, breast), to identify the disease status and prognoses, and possibly the effects of therapeutic treatment. First results are published [3-1]. Globus T, Ferrance J, Moskaluk C, Gelmont B, Bykhovski A (2018) "Sub-Terahertz Spectroscopic Signatures from micro-RNA Molecules in Fluid Samples for Ovarian Cancer Analysis". Case Rep Lit Rev 2(2): 100012, https://www.onjourn.com/case-reports-and-literature-review.php.
b) Application of sub-THz spectroscopy to study ovarian cancer from tissue samples. The characteristic spectroscopic features in absorption spectra, particularly the presence of absorption peaks near 13 cm-1 , which are very similar to the signature from 2 ovarian cancer lines samples (earlier published results [3-2]): T. Globus, I. Sizov, J. Ferrance, A. Jazaeri, J. Bryant, A. Moyer, B. Gelmont,
M. Kester and A. Bykhovski, "Sub-terahertz vibrational spectroscopy for microRNA based diagnostic of ovarian cancer". Converg. Sci. Phys. Oncol. 2 (2016) http://dx.doi.org/10.1088/2057-1739/2/4/045001, ), have been identified as this cancer indicators. We demonstrated that miRNA molecules dominate spectroscopic signatures of both cancer and normal control samples, although the miRNA profile is changing with cancer development. We also demonstrated that tissue samples heterogeneity, reflected by diverse spectral signatures, provides additional, very specific information that may be used for identification of cancer subtypes, clinical behavior or sensitivity to specific therapies. Because sub-THz spectroscopy provides information not reflected in tissue morphology, there is promise for this methodology as a supplemental analysis to traditional tissue diagnostic approaches. The paper has been submitted for publication [3-3]. T. Globus, C.Moskaluk, P. Pramoonjago, B. Gelmont, A. Moyer, A. Bykhovski, J. Ferrance, "Sub-Terahertz Vibrational Spectroscopy of Ovarian Cancer and Normal Control Tissue for Molecular Diagnostic Technology." submitted for publication to
Cancer Biomarkers Journal. Tracking number 18-2120).
Innovation and New Capabilities.
The recent introduction of new sources and detectors operating in the sub-THz frequency range has allowed Vibratess to take advantage of this opportunity to design and build instruments for multiple applications of vibrational resonance spectroscopy for molecular bio-sensing and identification of specific resonance features, molecular fingerprints in absorption spectra of molecules or entire biological cells.
Vibratess has developed and built instruments for characterization of ultra-small amounts of sample materials in solution with high sensitivity, high spectral and spatial resolution. The first instruments combined spectroscopic functions with imaging of sample materials that is important for applications in research laboratories. High sensitivity of these instruments and spatial resolution ~ 10-20 microns became possible via a novel solution to the fundamental problem of improving THz radiation coupling to biomolecules based on a local electro-magnetic field enhancement through the use of metal array sample holders. All Vibratess instruments are operating at room temperature between 315 and 490 GHz with a spectral resolution better than 3
GHz (~0.03 cm-1 ). The latest version of our spectrometer is a rather small, simple and easy to use instrument with a spectral resolution of 1 GHz or better The instruments have been intensively used for more than 6 years and tested for sensitivity, accuracy and reproducible results. The measurements procedures have been optimized, the required optimal amount of sample material and concentration in liquids have been found, and software for using the instruments have been developed.
New molecular diagnostic technology and instrumentation were introduced by Vibratess for accurate and predictive cancer detection, and identification of specific subsequent treatments that help identify the biology involved in cancer development and progression and promise to improve patient outcomes Vibratess' emerging technology is based on unique combined experimental and computational approach to predict, verify and validate the results of measurements and facilitate their analysis. We develop computational modeling techniques to study THz spectroscopic signatures of large macromolecules of DNAs, RNAs, proteins and other molecular components of cells using energy minimization, normal mode analysis and Molecular Dynamic (MD) approaches. Combining simulations with experimental techniques improves analysis and understanding of measured spectra and even provides predictive capabilities for modeling.
Using this combined approach, Vibratess introduced subTHz spectroscopy technology with high spectral and spatial resolution as an optical, label-free and reagent-free highly innovative approach to improve molecular and cellular characterization. THz spectroscopy is the only technique capable of directly detecting the weakest hydrogen bonds and other non-bonded interactions between atoms within biological molecules. The reality of the observed resonances in absorption spectra of biological molecules due to hydrogen bonds vibrations has been confirmed by computational modeling that demonstrated the existence of long lasting relaxation processes within molecules resulted in detectable absorption peaks.
([5-1]. T. Globus, I. Sizov and B. Gelmont, Sub-THz specific relaxation times of hydrogen bond oscillations in E.coli thioredoxin. Molecular dynamics and statistical analysis. The Royal Society of Chemistry 2014, Faraday Discuss. 2014, 171, 1–15 | 3. DOI: 10.1039/c4fd00029c ). Multiple resonances due to low energy vibrational modes within biological macromolecules and bio-cells have been unambiguously demonstrated in the sub-THz frequency range in agreement with the theoretical prediction (see Fig.1 for illustration [5-2]).
Thus, unique spectroscopic signatures do exist in absorption spectra of biological materials in this frequency range and are available for sensitive, specific characterization of bio-molecules and species. The fast and inexpensive spectroscopy technique provides useful genetically relevant information using less than nanogram of sample material. The results have been published. The demonstrated results of measurements and analysis are leading to multiple potential applications of a new technology including Oncology, where cancer is studied at the molecular level. These technology developments will move researchers toward early detection, diagnosis, prognosis, and therapeutic treatment of cancers and other diseases that are currently difficult to detect (Fig 2). Our optical sub-Terehertz spectroscopy technology does not require the extraction of microRNA and at the same time is solving important problems of their visualization and quantification, since the presence of microRNAs in serum or other liquid is revealed as a set of absorption peaks at specific frequencies and with their intensities specific to particular disease and it stage.
Fig.1. Absorption spectrum of protein thioredoxin from E. coli: MD simulation and experimental results using Vibratess spectroscopic sensor [5-2].
Fig. 2. Absorbance of Cancer S-line in isopropyl alcohol (brown) & normal control (green)
Vibratess produces the major breakthrough in subTHz Spectroscopic Sensing of Bio-materials
To be able to resolve vibrational modes from biological molecules and organisms, the spectra have to be measured with a relatively good spectral resolution that is adequate to the widths of spectral features. To date, most THz spectroscopy has been done at frequencies above 1 THz, for relatively small biomolecules. Full exploitation of THz vibrational spectroscopy is still impeded because of the absence of spectroscopic systems that simultaneously satisfy all the requirements of good spectral and spatial resolution, along with high sensitivity and room temperature operation. In addition, a simple sample preparation procedure, good reproducibility, and user-friendly operation are also important for widespread adoption of this technique.
Vibratess, LLC is a start up, women-owned small business company, orginazed in 2007 as a spin-off from the University of Virginia to develop and commercialize a novel low-THz resonance spectroscopy technology for detecting, identifying and characterizing biological macromolecules and microorganisms. Since then, a Sub-THz spectroscopic sensor with high sensitivity, good spectral resolution, and spatial resolution below diffraction limit has been developed.
Frequency domain sub-THz spectroscopic bio-sensor system (Vibratess, LLC)
This novel constant wave, frequency-domain instrument is based on a patented strong local enhancement of the electro-magnetic field, thus allowing increased coupling of the THz radiation with the sample biomaterials. The invention uses the discontinuity edge effect and the extraordinary transmission of a sub-wavelength-slit conductive structure [B. Gelmont, T. Globus, et al “Method of Local Electro-Magnetic Field Enhancement of Terahertz (THz) Radiation in Sub Wavelength Regions and Improved Coupling of Radiation to Materials through the Use of the Discontinuity Edge Effect”, Patent No. 8,309,930 issued November 26. 2012].
Absorption spectrum of protein thioredoxin from E. coli: MD simulation and experimental results as measured using Vibratess’ spectroscopic sensor.
For scientific applications, our new instrument opens opportunities for better understanding the physics of interaction between THz radiation and bio material and time scales of intramolecular dynamics relaxation processes. Sub-Thz spectroscopy coupled with theoretical prediction can become a powerful tool for the investigation of the molecular structure and possible biological function.
Vibratess has a strong theoretical group. It focuses on two aspects: 1. Using molecular dynamics (MD) and other computational tools to predict, to verify and confirm experimental signatures from known and unknown biological objects. We have significantly improved the predictive capability of MD simulation for proteins and developed a novel approach for a computational modeling of THz vibrational spectra from biological macromolecules. In parallel with experimental characterization, simulated absorption spectra have been received for DNA molecules from different strains of E.coli, from protein Thioredoxin, and from artificial engineered molecular structures - DNA monocrystals. Vibratess has clearly demonstrated that the frequencies of resonances and absorption spectra patterns for different biomaterials are unique. Comparison between simulated and experimental spectra enhances understanding of the physics of intramolecular atomic motions and dynamics relaxation processes in biological macromolecules. 2. Optimizing experimental setups to improve interaction between THz radiation and biological materials, to more effectively collect and transfer the information.
Validation: Computational Modeling
Vibratess is constantly working on refinements and improvements, including customized biosensors, designed for specific wavelengths and bio-targets. Additionally, we are developing a novel, inexpensive microfluidic chip with multiple ports to allow precise control of the sample, as well as the introduction of reagents. The fluid can be blood, to detect cancer cells and signaling molecules, for example, or an environmental water sample. Ultimately, our goal is to reduce the system to a portable, field-ready version. Vibratess also continues experiments with new biomaterials. Most recently, Vibratess successfully identifies the unique spectroscopic signature or “fingerprint” of ovarian cancer cells in cell culture. The system can detect and identify a single cancer cell by interrogation of specific resonances caused by intra-molecular motions within the cell. With the addition of the novel microfluidic sample device that is currently in development, the system can identify individual cells in liquids, detect signaling molecules circulating in blood, and assess therapeutic responses in cancer or other targeted cells. The multi-port device is also expected to expedite testing and development of treatment modalities. Current work is aimed at identifying signatures for normal vs. malignant cells.
The Vibratress system achieves good spectral resolution, spatial resolution below diffraction limit, and is highly sensitive. It requires only nanograms of material for fast (approximately 10 minute) sample characterization at room temperature with reproducible results. Sample preparation is simple, with no reagents or labels. Since there is low disturbance from water or other solvents, detection in solution is possible. Vibratess offers a sturdy system that is relatively small and inexpensive and operates with user-friendly software. The system, unlike others, has actual experimental results to demonstrate its capabilities. The first patent has issued on this technology, which has been further validated by continuing government support; two more patents are pending. Vibratess carefully manages its intellectual property portfolio, which includes a high level of expertise.
The Vibratess system has broad applications:
1) Creating reference libraries of pathogens, biohazards, diseased cells, contaminants and so on for rapid field and lab testing. Faster identification means a faster response time.
2) Providing an in-house tool for hospitals to rapidly analyze bio-cell and tissue to screen, diagnose, and evaluate disease states, such as cancer.
3) Enhancing drug discovery, biomaterial characterization and pharmokinetics studies; the multi-port device allows evaluation of a target’s response to various compounds; process control, product inspection.
4) Expediting research into cellular and biological processes, including mechanisms producing disease; real-time monitoring of biological processes – folding-unfolding, and conformational changes, for example.
5) Improving national security by the rapid detection of chemical and biological agents, and environmental contaminants.
6) Promoting research and environmental science in universities, national laboratories, and the commercial sector
Vibratess is entering a young market. The first commercial THz spectrometers were introduced less than a decade ago. Since then, the market has grown to more than $10 million in annual sales, with more than half a dozen vendors. These systems however require relatively large amounts of material, and in most cases fail to produce sufficient spatial and spectral resolution to detect and characterize the unique fingerprints of biomaterials.
The sub-THz vibrational spectroscope is an advanced, unique diagnostic, screening and research tool. The overall goal for Vibratess is to have its spectroscopic sensor as ubiquitous and taken for granted as a microscope. This system will provide unique possibilities in detection, separation, classification and identification applications. The system is a stand-alone product, but Vibratess also is developing and refining its component parts, such as biosensors and sample holders-chips, to sell as customized devices that interface with existing THz spectroscopy systems.
Oncology: Vibratess brings a vital new tool for diagnosis, prognosis, treatment, and early detection. The timing is just right, since cancer is being studied at the molecular level to find diagnostic and therapeutic targets. The annual global market for next-generation cancer diagnostics was $776 million in 2010 and growing at a compound annual growth rate of 47%.
Antibiotics: Antibiotics generate sales of over $42 billion and grow at about 4% a year. A single drug family, cephalosporin, makes up $11.9 billion of that, with 28% of the market. However, as patents expire and antibiotic resistance becomes an increasing problem, there is a trend to first identify pathogens with lab tests instead of routinely prescribing antibiotics based on symptoms. Antibiotic-resistant organisms have led to a demand for new types of antibiotics that more precisely target pathogens and reduce side effects. Presently, this is laborious and uncertain work. Vibratess offers more rapid diagnosis and a new technique to study targets and evaluate the activity of candidate therapeutic compounds.
National Security: Detection of biothreats is a high priority. Vibratess has the potential to identify pathogens in minutes, rather than days, as is standard now. Recently ARO and ECBC have launched a multi-disciplinary research program under the support of the U.S. Defense Threat Reduction Agency (DTRA) that seeks to develop new devices that can obtain THz signatures from target bio-molecules. These government agencies and national laboratories will be priority customers for Vibratess.
Water-borne and other contaminants: The Centers for Disease Control (CDC) has created a special agency to address the global problem of prevention and control of diseases caused by zoonotic, vector-borne, food-borne, water-borne, mycotic, and related infections. Testing household water and bodies of water in the environment must be rapid and reliable. Appropriate detection methods are also needed to obtain clinical and environmental samples from the air and ground to detect parasitic and other contaminants.