This PDF file contains the front matter associated with SPIE Proceedings Volume 11938, including the Title Page, Copyright information, Table of Contents, and Conference Committee listings.

Imaging mass spectrometry (IMS) has emerged as a powerful technology in the pharmaceutical industry with many applications to enhance drug discovery and development. In order to use IMS to support drug development, IMS researchers must develop, validate, and establish best practices to assure data integrity to adequately support new drug registrations. The Imaging Mass Spectrometry Society (IMSS) initiated a project to examine how MS is currently being used in the industry and partnered with the Japan Association for Imaging Mass Spectrometry (JAIMS) to conduct a thorough survey of IMS scientists, data analysts/users, project managers/leaders, and pharmaceutical compliance specialists who currently use IMS techniques in support of their drug discovery and development efforts. The survey asked detailed questions related to technical aspects of: 1. sample collection & preparation; 2. qualitative and quantitative sample analysis; 3. examination of parent drug, metabolites and biomarkers; 4. data acquisition, reporting and integrity; 5. General applications; and 6. regulatory concerns. This international survey was conducted online through Survey Monkey in both English and Japanese in September of 2020, with a goal of beginning efforts to establish best practices for the wide acceptance of IMS data for use in regulated drug development industry. This oral presentation will discuss the preparation and conduct of the survey, survey results, and to provide the audience with an approach to developing surveys to help establish best practices for the application of other drugimaging technologies in the Pharmaceutical Industry.

We studied quantitative evaluation of skin erythema using dorsum of hand images of photodynamic therapy (PDT) patients before and after the exposure to sunlight. While PDT is used as a minimally invasive cancer treatment, there is a problem of photosensitivity caused by residual photosensitizer in the skin. To prevent this problem, a dark hospitalization of two weeks in a room with less than 500 lx is prescribed in the case of Npe6 in Japan. After 2 weeks of a dark hospitalization, dorsum of hand is exposed to sunlight for 5 minutes, and the photosensitivity is visually judged to determine the end of a dark hospitalization, but this method has the problem of being qualitative and invasive. The purpose of this study was to quantitatively evaluate cutaneous photosensitivity by analyzing dorsum of hand images of PDT patients before and after the exposure to sunlight using three colorimetric systems. Dorsum of hand images before and after sunlight exposure of nine PDT patients were taken along with color scales. Image processing software was used to extract the sRGB values of the sunlight-exposed area and color scale from the acquired images as histograms. The sRGB values of the color scale of the images were used to correct the imaging conditions and to compare the images before and after sunlight exposure in three color systems: sRGB, CIELAB, and HSV color system. We suggest that the G values of sRGB has the possibility of quantitative evaluation of erythema and photosensitivity.

Here we present novel cloud-connected theranostic medical laser platform specifically designed for activating and simultaneously monitoring multi-component oncological treatment processes. It may incorporate multiple wavelengths for inducing therapeutic effect or monitoring treatment in real-time. The same low-invasive optical probers can be used for treatment and monitoring. We believe that this theranostic laser platform will allow clinicians to develop improved treatment outcomes for cancer patients that may be based on machine learning and AI in the future.

Head and neck squamous cell carcinomas (HNSCCs) have high levels of chromosomal instability and epidermal growth factor receptor (EGFR) overexpression, both of which drive their tumorigenesis. While drug treatment targeting the extracellular domain of EGFR has shown some success, mutations and alternate intracellular pathways contribute to therapeutic resistance. Therefore, a dynamic in vivo method to monitor binding and downstream cell signaling is warranted. Previous work has demonstrated that paired-agent imaging (PAI) is a powerful tool to quantify extracellular EGFR, and so this work extends the same principles to quantify intracellular protein target engagement. Here, in ovo models were used to grow human HNSCC xenografts – eggs were windowed to reveal the chorioallantoic membrane (CAM) of chicken embryos and tumors were implanted on its surface. A fluorescent cocktail of both intracellular and extracellular, targeted and untargeted agents (four agents total) was intravenously injected and multispectral imaging was performed over two hours. To isolate the relative quantities of each agent, a spectral fitting procedure was employed that accounted for the linear contributions of each fluorescent agent and autofluorescence, and the non-linear absorbing contributions of oxy- and deoxyhemoglobin. This unmixing was performed on a pixel-by-pixel basis to generate distribution maps of each individual dye, and then motion correction was done, followed by a convolution correction to account for delivery differences. Results demonstrated successful unmixing of individual fluorophores such that a ratiometric calculation could be applied to extract both intracellular and extracellular binding potential (BP), which is proportional to EGFR concentration.

We present ongoing work to develop and verify a measurement technology using diffuse reflectance spectroscopy and fluorescence spectroscopy to measure in-vivo concentrations of intact and dissociated porphysomes. Porphysomes (PS) are multifunctional liposome-like nanoparticles composed of porphyrin-lipid subunits that exhibit structure-dependent optical quenching. They are attractive as agents for photothermal and photodynamic therapy but methods are required for measuring their concentration in the intact and dissociated states for treatment dosimetry and guidance. Diffuse reflectance spectroscopy is used here to measure concentrations of intact and dissociated (unquenched) PS using their unique absorption spectra, and also to correct the fluorescence signal for the optical properties of the tissue. The latter is then used to quantify the fluorescence (QF) signal from dissociated PS, providing a more direct measure of photodynamic activity. Two experiments were performed for proof-of-concept. First, a liquid phantom was used to verify measurements taken with a combined reflectance-fluorescence probe. This demonstrated that the reflectance signal was sensitive to the relative proportion of intact and dissociated PS and that the QF signal contained information about the PS unquenching. Second, a mouse model was used to demonstrate that the drop in the reflectance signal at the PS absorption peak wavelength and the fluorescence signal are consistent with Positron Emission Tomography (PET) measurements of copper-64 labeled PS.

Recently, sonodynamic therapy (SDT) is actively developed, and the reason is that it goes beyond the conventional photodynamic therapy (PDT). SDT demonstrates exceptional ability to treat deep-seated tumor. However, it is difficult to develop sonosensitizers used in SDT. Herein, we demonstrate dual stimuli-responsive theranostic nanosonosensitizers using biocompatible extracellular vesicles (EVs), a class of naturally occurring nanoparticles. With this nanosonosensitizers, we can achieve both high-cancer-targeting and biocompatible properties. Indocyanine green (ICG), used as an sonosensitizer and a photoacoustic (PA) imaging agent, was loaded into EV with paclitaxel (PTX) and sodium bicarbonate (SBC) to achieve pH-responsive PA image-guided chemo-SDT. The SBC trapped in the EV generates CO2 bubbles and causes the EV to burst. That's why SBC-, ICG-, PTX-equipped EVs [ICG/PTX] can efficiently release drugs in response to acidic pH in the endo/lysosomes. The photostability and cellular uptake of ICG was improved by EVs. Further, the ICG-, PTX-loaded EVs and SBCEV( ICG/PTX) showed great anticancer activities against breast cancer cells. The effect of SBC-EV was shown through high resolution PA imaging, which showed that the SBC-EV is accumulated in tumor. SBC-EV (ICG/PTX) with insonification showed that inhibition of tumor growth is possible without toxicity. Thus, this study demonstrates that dual stimuli-responsive SBC-EV(ICG/PTX) holds a great potential for clinical translation of chemo-sonodynamic cancer theranostics.

A major challenge in the study of the physical mechanism behind needle-free jet injection (NFJI) systems is the lack of visualization techniques or tools to study the penetration and dispersion characteristics of the high-velocity microjet inside the skin tissue. Current imaging techniques used to assess skin permeation require a minimum temporal resolution of 1 ms, whereas a temporal resolution in the order of 100 μs is needed to study the penetration and dispersion phases of a NFJI system. In this study, we propose near-infrared (NIR) fluorescent and absorbance/transmission imaging techniques to achieve the temporal resolution required for visualization of NFJI studies. Fluorescent signals up to 2.5 mm could be obtained with a normal ex vivo porcine sample. However, absorbance/transmission imaging could not produce the required contrast in the normal ex vivo sample. Also, we apply a tissue clearing technique to improve the image acquisition for higher depth. In the cleared tissue sample, the lowest measured dimensional error was observed at a temporal resolution up to 50 and 10 μs for NIR fluorescent and absorbance/transmission imaging techniques, respectively.