Advances in mass spectrometry-based cancer research and analysis: from cancer proteomics to clinical diagnostics

Introduction: The last 20 years have seen significant improvements in the analytical capabilities of biological mass spectrometry. Studies using advanced mass spectrometry (MS) have resulted in new insights into cell biology and the aetiology of diseases as well as its use in clinical applications. Areas Covered: This review will discuss recent developments in MS-based technologies and their cancer-related applications with a focus on proteomics. It will also discuss the issues around translating the research findings to the clinic and provide an outline of where the field is moving. Expert Opinion: Proteomics has been problematic to adapt for the clinical setting. However, MS-based techniques continue to demonstrate potential in novel clinical uses beyond classical cancer proteomics

Autofluorescence lifetime augmented reality as a means for real-time robotic surgery guidance in human patients.

Due to loss of tactile feedback the assessment of tumor margins during robotic surgery is based only on visual inspection, which is neither significantly sensitive nor specific. Here we demonstrate time-resolved fluorescence spectroscopy (TRFS) as a novel technique to complement the visual inspection of oral cancers during transoral robotic surgery (TORS) in real-time and without the need for exogenous contrast agents. TRFS enables identification of cancerous tissue by its distinct autofluorescence signature that is associated with the alteration of tissue structure and biochemical profile. A prototype TRFS instrument was integrated synergistically with the da Vinci Surgical robot and the combined system was validated in swine and human patients. Label-free and real-time assessment and visualization of tissue biochemical features during robotic surgery procedure, as demonstrated here, not only has the potential to improve the intraoperative decision making during TORS but also other robotic procedures without modification of conventional clinical protocols

Indocyanine Green-based Glow Nanoparticles Probe for Cancer Imaging

Indocyanine green (ICG) is one of the FDA-approved near infra-red fluorescent (NIRF) probes for cancer imaging and image-guided surgery in the clinical setting. However, the limitations of ICG include poor photostability, high concentration toxicity, short circulation time, and poor cancer cell specificity. To overcome these hurdles, we engineered a nanoconstruct composed of poly (vinyl pyrrolidone) (PVP)-indocyanine green that is cloaked self-assembled with tannic acid (termed as indocyanine green-based glow nanoparticles probe, ICG-Glow NPs) for the cancer cell/tissue-specific targeting. The self-assembled ICG-Glow NPs were confirmed by spherical nanoparticles formation (DLS and TEM) and spectral analyses. The NIRF imaging characteristic of ICG-Glow NPs was established by superior fluorescence counts on filter paper and chicken tissue. The ICG-Glow NPs exhibited excellent hemo and cellular compatibility with human red blood cells, kidney normal, pancreatic normal, and other cancer cell lines. An enhanced cancer-specific NIRF binding and imaging capability of ICG-Glow NPs was confirmed using different human cancer cell lines and human tumor tissues. Additionally, tumor-specific binding/accumulation of ICG-Glow NPs was confirmed in MDA-MB-231 xenograft mouse model. Collectively, these findings suggest that ICG-Glow NPs have great potential as a novel and safe NIRF imaging probe for cancer cell/tumor imaging. This can lead to a quicker cancer diagnosis facilitating precise disease detection and management

Innovative Device for Indocianyne Green Navigational Surgery

Dynamic reality has been integrated into developing surgical techniques, with the goals of providing increased intraoperative accuracy, easier detection of critical anatomical landmarks, and better general results for the patient. Enhancement of the reality in surgical theaters using single or multi sensorial augmenters (haptic, thermic and visual) has been reported with various degrees of success. This paper presents a novel device for navigational surgery and ancillary clinical applications based on the fluorescent properties of Indocyanine Green (ICG), a safe, FDA-approved dye that emits fluorescence at higher wavelengths than endogenous proteins. The latest technological developments and the aforementioned convenient quantum behavior of ICG allow for its effective identification in tissues by means of a complementary metal-oxide semiconductor (CMOS) infrared camera. Following fundamental research on the fluorophor in different biological suspensions and at various concentrations, our team has built a device that casts a beam of excitation light at 780nm and collects emission light at 810-830nm, filtering ambient light and endogenous autofluorescence. The emission light is fluorescent and infrared, unlike visible light. It can penetrate tissues up to 1.6cm in depth, providing after digitization into conventional imaging anatomical and functional data of immense intra-operative value

The Development of Hyaluronan-Based Contract Agents for the Intraoperative Detection of Pancreatic Tumors

Pancreatic ductal adenocarcinoma is highly lethal and surgical resection is the only potential curative treatment for the disease. Tumor-specific intraoperative fluorescence imaging could improve staging and surgical resection, thereby improving prognosis. In the first study, hyaluronic acid derived NPs with physico-chemically entrapped indocyanine green, termed NanoICG, were utilized for intraoperative near infrared fluorescence detection of pancreatic cancer. NanoICG accumulated significantly in an orthotopic pancreatic ductal adenocarcinoma model with safety profile both in vitro and in vivo. To maximize tumor signal, while minimizing signal in healthy pancreas and RES capture of macromolecules, in the next study, we describe the rational development of a series of hyaluronic acid (HA) conjugates that vary in molecular weight and are conjugated to near-infrared fluorescent (NIRF) dyes that have differences in hydrophilicity, serum protein binding affinity, and clearance mechanism. We systematically investigated the roles of each of these properties on tumor accumulation, relative biodistribution, and the impact of intraoperative imaging of orthotopic, syngeneic pancreatic cancer. Overall, each HA-NIRF conjugate displayed intra-pancreatic tumor enhancement compared to uninvolved pancreas at 24 and 96 h. Regardless of HA molecular weight, Cy7.5 conjugation directed biodistribution to the liver, spleen, and bowels. Conjugation of IRDye-800 to 5 and 20 kDa HA resulted in low liver and spleen signal, while preserving tumor contrast enhancement up to 14-fold compared to healthy pancreas. When IRDye800 was conjugated to 100 kDa HA, the conjugate preferentially distributed to RES organs. When assessing the imaging efficacy of HA-based conjugates in hepatic metastases, those that accumulated to the liver utmost (HA100k-Cy7.5, HA100k-IRDye800, NanoICG) turned to aid the identification of hepatic malignancy with hypo-contrast. These studies demonstrate that by tuning HA molecular weight and the physicochemical properties of the conjugated moiety, in this case a NIRF probe, peritoneal biodistribution can be substantially altered to achieve optimized delivery to tumors with robust contrast enhancement for intraoperative imaging to abdominal tumors. Aside from assisting the accurate delineation of primary tumor, HA-NIRF conjugates demonstrated potential for identification of occult metastases in the intraoperative setting, as a versatile tool for accurate staging

Curcumin: A Multi-dimensional Approach to Pancreatic Cancer Targeting Cell Death and Exosomes

Pancreatic cancer is currently one of the most difficult diseases to treat due to difficulty of detection and the aggressive nature of the disease. In addition, pancreatic cancer has the highest mortality rates compared to other cancer types. These mortality rates are attributable in part to increasing resistance to cancer therapy. Cancer therapy resistance is caused by adaptations that favor survival within cancer cells and their environment, termed the tumor microenvironment. Intracellular adaptations include the overexpression of resistance-linked genes, such as the inhibitor of apoptosis (IAP) family of proteins and overall resistance to cell death. Adaptations in the tumor microenvironment include altered intercellular vesicular signaling through exosomes, resulting in tumor growth and progression. However, recent studies have shown that exosomes can also be used as a delivery mechanism for drugs with poor bioavailability, thus providing a therapeutic advantage for these compounds. Currently, researchers are moving toward a multi-dimensional approach to pancreatic cancer therapy that incorporates compounds that target crucial players in chemotherapy resistance and in the tumor microenvironment, such as exosomes. Our studies are centered on the anti-cancer properties of curcumin, a turmeric derivative, on these intracellular and intercellular resistance mechanisms. The long term goal of this research is to determine the mechanisms by which curcumin modulates intracellular pathways related to pancreatic cancer survival and therapy resistance and exosome composition and release to improve the understanding of pancreatic cancer pathology and support the development of novel therapeutic approaches for pancreatic cancer patients. The specific objective of this research was to determine curcumin’s role in modulating intracellular proteins imperative for pancreatic cancer chemotherapy resistance such as the IAP proteins. Moreover, this research addressed the effects of curcumin on exosome release and function, specifically in the context of delivery to recipient pancreatic cancer cells. We have established that curcumin reduces expression of the IAPs in pancreatic cancer cells, inhibiting their survival and growth. Furthermore, curcumin not only attenuates pro-survival signaling through exosomes, but also itself carried within the nanovesicles and delivered to recipient pancreatic cancer cells, resulting in pancreatic cancer cell death

Fluorescence-Guided Surgery and Fluorescence Laparoscopy for Gastrointestinal Cancers in Clinically-Relevant Mouse Models

There are many challenges that face surgeons when attempting curative resection for gastrointestinal cancers. The ability to properly delineate tumor margins for complete resection is of utmost importance in achieving cure and giving the patient the best chance of prolonged survival. Targeted tumor imaging techniques have gained significant interest in recent years to enable better identification of tumor lesions to improve diagnosis and treatment of cancer from preoperative staging modalities to optimizing the surgeon’s ability to visualize tumor margins at the initial operation. Using unique characteristics of the tumor to fluorescently label the tissue can delineate tumor margins from normal surrounding tissue, allowing improved precision of surgical resection. In this paper, different methods of fluorescently labeling native tumor are discussed as well as the development of fluorescence laparoscopy and the potential role for fluorescence-guided surgery in the treatment of gastrointestinal cancers