Dynamic Optical Contrast Imaging Research Articles

Abstract B013: Development of oropharyngeal tumor organoids for disease modeling and high throughput drug screening

Abstract Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide with approximately 550,000 new cases diagnosed each year. The oropharynx is one of the most common sites of occurrence, and disease in this region most often presents in the palatine tonsil, followed by the base of tongue and soft palate. Human papilloma virus (HPV) is a common cause of HNSCC, giving rise to a distinct subset of tumors with a different clinical presentation, disease course, and response to treatment. In order to further characterize HPV+ HNSCC tumors and their therapy response, we have leveraged our high throughput 3D tumor organoid screening platform. Our approach entails seeding tumor cells in extracellular matrix gels deposited around the perimeter of tissue culture wells in ring-like structures, with screening results available within a week from surgery (Phan et al, 2019; Al Shihabi et al, 2022; Nguyen et al, 2022; Al Shihabi et al 2023). We particularly focused on tonsil tumors, and develop organoids from both HNSCC and contralateral normal tonsil tissue. Thus far, we have procured tonsil samples from 31 patients to develop organoids, perform high throughput drug screening, and immunohistochemical characterization including H&E staining, Ki67, p16 and p53 immunohistochemical staining. We have leveraged dynamic optical contrast imaging (DOCI), a label-free tissue autofluorescence imaging method that is being investigated clinically (Kim et al, 2017), to assist in distinguishing tumor organoids from normal tissue organoids. Lastly, given the importance of radiation as a treatment modality for HPV+ HNSCC, we have also optimized a chemoradiation pipeline to test possible combinatorial effects. Taken together, our results demonstrate that 3D organoids of HNSCC generated from patient samples can successfully preserve native tissue characteristics and can be used for high throughput drug screening and radiation de-escalation testing, affording critical insight into the management of HNSCC in patients. Citation Format: Krishna K. Bommakanti, Luda Lin, Yazeed Alhiyari, Andreanne Sannajust, Brandon Mo, Lauran Evans, Peyton Tebon, Maie St. John, Alice Soragni. Development of oropharyngeal tumor organoids for disease modeling and high throughput drug screening [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr B013.

Real-Time Intraoperative Detection of Margins for Oral Cavity Squamous Cell Carcinoma.

Obtaining negative surgical cancer margins is the strongest predictor for the long-term survival of oral cavity squamous cell carcinoma patients. To verify that the tumor has been completely removed, surgeons rely on pathologic evaluation of frozen sections to determine surgical margins, which can be time-consuming and subjective. Herein, we detail the real-time intraoperative use of dynamic optical contrast imaging (DOCI), a novel imaging modality that rapidly distinguishes head and neck cancer from healthy adjacent tissues based on fluorescence decay information from spectral bands in the UV-VIS range. Analysis of DOCI revealed microscopic characterization sufficient for tissue type identification consistent with histology (p < .05). DOCI delivers a clinically relevant tool that may better inform and drive precision surgery, directly impacting surgical outcomes and improving overall survival for our patients.

Label-free, real-time detection of perineural invasion and cancer margins in a murine model of head and neck cancer surgery

Surgical management of head and neck cancer requires a careful balance between complete resection of malignancy and preservation of function. Surgeons must also determine whether to resect important cranial nerves that harbor perineural invasion (PNI), as sacrificing nerves can result in significant morbidity including facial paralysis. Our group has previously reported that Dynamic Optical Contrast Imaging (DOCI), a novel non-invasive imaging system, can determine margins between malignant and healthy tissues. Herein, we use an in vivo murine model to demonstrate that DOCI can accurately identify cancer margins and perineural invasion, concordant with companion histology. Eight C3H/HeJ male mice were injected subcutaneously into the bilateral flanks with SCCVIISF, a murine head and neck cancer cell line. DOCI imaging was performed prior to resection to determine margins. Both tumor and margins were sent for histologic sectioning. After validating that DOCI can delineate HNSCC margins, we investigated whether DOCI can identify PNI. In six C3H/HeJ male mice, the left sciatic nerve was injected with PBS and the right with SCCVIISF. After DOCI imaging, the sciatic nerves were harvested for histologic analysis. All DOCI images were acquired intraoperatively and in real-time (10 s per channel), with an operatively relevant wide field of view. DOCI values distinguishing cancer from adjacent healthy tissue types were statistically significant (P < 0.05). DOCI imaging was also able to detect perineural invasion with 100% accuracy compared to control (P < 0.05). DOCI allows for intraoperative, real-time visualization of malignant and healthy tissue margins and perineural invasion to help guide tumor resection.

Ex vivo hypercellular parathyroid gland differentiation using dynamic optical contrast imaging (DOCI).

Primary hyperparathyroidism, often caused by a single adenoma (80-85%) or four-gland hyperplasia (10-15%), can lead to elevated parathyroid hormone (PTH) levels and resultant hypercalcemia. Surgical excision of offending lesions is the standard of care, as the removal of pathologic adenomas reduces PTH and calcium values to baseline. The small size, variable location, and indistinct external features of parathyroid glands can make their identification quite challenging intraoperatively. Our group has developed the dynamic optical contrast imaging (DOCI) technique, a novel realization of dynamic temporally dependent measurements of tissue autofluorescence. In this study, we evaluated the efficacy of using the DOCI technique and normalized steady-state fluorescence intensity data for differentiating types of human parathyroid and thyroid tissues. We demonstrate that the DOCI technique has the capability to distinguish normal parathyroid tissue from diseased parathyroid glands as well as from adjacent healthy thyroid and adipose tissue across 8 different spectral channels between 405nm-600nm (p<0.05). Patient tissue DOCI data was further analyzed with a logistic regression classifier trained across the 8 spectral channels. After computer training, the computer-aided identification was able to accurately locate hypercellular parathyroid tissue with 100% sensitivity and 98.8% specificity within the captured DOCI image.

A Tool to Locate Parathyroid Glands Using Dynamic Optical Contrast Imaging.

Identification of parathyroid glands and adjacent tissues intraoperatively can be quite challenging because of their small size, variable locations, and indistinct external features. The objective of this study is to test the efficacy of the dynamic optical contrast imaging (DOCI) technique as a tool in specifically differentiating parathyroid tissue and adjacent structures, facilitating efficient and reliable tissue differentiation. Prospective study. Both animal and human tissues were included in this study. Fresh specimens were imaged with DOCI and subsequently processed for hematoxylin and eosin (H&E) stain. The DOCI images were analyzed and compared to the H&E results as ground truth. In both animal and human experiments, significant DOCI contrast was observed between parathyroid glands and adjacent tissue of all types. Region of interest analysis revealed most distinct DOCI values for each tissue when using 494 and 572 nm-specific band pass filter for signal detection (P < .005 for porcine tissues, and P=.02 for human specimens). Linear discriminant classifier for tissue type prediction based on DOCI also matched the underlying histology. We demonstrate that the DOCI technique reliably facilitates specific parathyroid gland localization. The DOCI technique constitutes important groundwork for in vivo precision endocrine surgery. 4 Laryngoscope, 131:2391-2397, 2021.

Abstract A01: Clinical detection of melanoma via endogenous fluorophore lifetime imaging

Abstract Objective: The 5-year survival for cutaneous malignant head and neck melanoma is very poor (17%). As proof of principle, we detect variations in endogenous fluorophore lifetimes by illuminating tissue with pulsed ultraviolet (UV) light. We have previously shown that dynamic optical contrast imaging is capable of delineating tumor margins. Our aim herein was to acquire clinical and intraoperative multispectral images that permit selection of regions of interest (ROI) to reliably and noninvasively perform a rapid optical biopsy in the patient. Methods: Patient cases of cutaneous melanoma were acquired with pulsed UV illumination utilizing a 6-diode in-series system. Emitted fluorescence was detected with a nanosecond gated CCD detector. The acquired signal is transformed into a relative difference value representing tissue fluorophore lifetime. Results: From (n=4) patient cases we acquired DOCI images both pre- and post-resection leading to a database of in vivo relative lifetime values that align to histology sections from corresponding locations via pathology. The spectral features of our images enabled classification of clinician-determined regions of interest in DOCI images in order to establish measures of internal and external validity of our novel device. Conclusion: Our device augments native tissue contrast via wide-field imaging to produce spatially resolved and pathology-specific maps of tissue. Our database of histology-matched real-time images is aimed to permit intraoperative guidance for tumor margin delineation in dermatologic surgery. Citation Format: Peter A. Pellionisz, Yong Hu, Jenny Kim, Warren Grundfest, Maie A. St. John. Clinical detection of melanoma via endogenous fluorophore lifetime imaging [abstract]. In: Proceedings of the AACR Special Conference on Melanoma: From Biology to Target; 2019 Jan 15-18; Houston, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(19 Suppl):Abstract nr A01.

Preoperative and Intraoperative Methods of Parathyroid Gland Localization and the Diagnosis of Parathyroid Adenomas.

Accurate pre-operative determination of parathyroid glands localization is critical in the selection of minimally invasive parathyroidectomy as a surgical treatment approach in patients with primary hyperparathyroidism (PHPT). Its importance cannot be overemphasized as it helps to minimize the harmful side effects associated with damage to the parathyroid glands such as in hypocalcemia, severe hemorrhage or recurrent laryngeal nerve dysfunction. Preoperative and intraoperative methods decrease the incidence of mistakenly injuring the parathyroid glands and allow for the timely diagnosis of various abnormalities, including parathyroid adenomas. This article reviews 139 studies conducted between 1970 and 2020 (49 years). Studies that were reviewed focused on several techniques including application of carbon nanoparticles, carbon nanoparticles with technetium sestamibi (99m Tc-MIBI), Raman spectroscopy, near-infrared autofluorescence, dynamic optical contrast imaging, laser speckle contrast imaging, shear wave elastography, and indocyanine green to test their potential in providing proper parathyroid glands’ localization. Apart from reviewing the aforementioned techniques, this study focused on the applications that helped in the detection of parathyroid adenomas. Results suggest that applying all the reviewed techniques significantly improves the possibility of providing proper localization of parathyroid glands, and the application of indocyanine green has proven to be the ‘ideal’ approach for the diagnosis of parathyroid adenomas.

Method for motion artifact compensation in dynamic optical contrast imaging

It has always been challenging for surgeons to localize offending lesions because of their small size, variable locations and indistinct external features while surgical interventions. Traditional methods include palpating edges and following analysis of biopsied tissues, which are limited in experienced hands or time consuming. Our group was able to innovate an intraoperative imaging system which generates wide-field tissue contrast images using relative differences in autofluorescence lifetimes at the speed of a few seconds per frame. In our technique, due to the relatively long gating time of each frame and intervals between frames, human movements are inevitably reflected in the images, yielded low success rate and required multiple re-runs to achieve satisfying results. This paper presents a way to minimize the influence of movements.

Clinical Translation and Optimization of Dynamic Optical Contrast Imaging for Intraoperative Surgical Margin Assessment

Clinical Translation and Optimization of Dynamic Optical Contrast Imaging for Intraoperative Surgical Margin Assessment

Intraoperative Adjunct Methods for Localization in Primary Hyperparathyroidism.

Primary hyperparathyroidism (pHPT) is a frequently seen endocrine disease, and its main treatment is surgery. In the majority of pHPT, the disease involves only a single gland, and the majority of the pathological glands can be determined by preoperative localization methods.In addition to preoperative localization studies in parathyroidectomy, the use of adjunct methods to improve intraoperative localization in order to increase success of surgery is becoming widespread. These methods include different approaches, mainly intraoperative parathyroid hormone (PTH) measurement, followed by intraoperative gamma probe application, intraoperative ultrasonography, parathyroid imaging with methylene blue, and frozen section examination. Recently, especially promising new imaging methods have been described in the literature with various optical technologies to increase the localization of the parathyroid glands and to evaluate their viability. These methods include parathyroid imaging with autofluorescence, indocyanine green imaging with autofluorescence, autofluorescence imaging with methylene blue, autofluorescence imaging with 5-aminolevulinic acid, optical coherence tomography, laser speckle contrast imaging, dynamic optical contrast imaging, and Raman spectroscopy. Currently, minimally invasive parathyroidectomy has become the standard treatment for selected pHPT patients with the aid of preoperative imaging and intraoperative auxiliary methods . The aim of the present study was to evaluate the routinely used new promising intraoperative adjunct methods in pHPT.

Abstract C83: Dynamic Optical Contrast Imaging (DOCI): Oral screening in the underserved

Abstract Objective: Oral and head and neck squamous cell carcinoma (OSCC) is the sixth most common cancer and disproportionately affects racial minorities via increased incidence and poorer rate of survival. Stage at presentation is one of the strongest prognostic indicators in new diagnoses; patients who present with advanced stage have 5-year survival rates of 42%, compared to patients who present at stage I with 5-year survival rates of 79%. Because five-year survival is directly related to stage at diagnosis, early detection of precancerous OSCC lesions could greatly reduce morbidity and improve patient survival. We aim to demonstrate the utility of a novel tool (Dynamic Optical Contrast Imaging; DOCI) in reliably and accurately delineating tumor tissue from surrounding normal tissues for earlier patient screening. Methods: OSCC specimens and surrounding tissues from the surgical bed were collected; fluorescence decay images were acquired using our DOCI system. Samples (55 patients) were subsequently processed and histologically assessed by head and neck pathologists. Mean relative fluorescence decay signatures were calculated for tumor, fat, muscle and collagen tissues. Statistical analyses were performed using the Wilcoxon signed rank test. Results: Statistical significance (P&amp;lt;.05) between muscle and tumor was established for 10 emission wavelengths, collagen and tumor for 8 emission wavelengths, and between fat and tumor for 2 wavelengths. Conclusion: We demonstrate the utility of DOCI in distinguishing OSCC from surrounding normal tissue. This novel screening tool would permit noninvasive tissue identification in underserved populations by reducing the stage of tumor presentation at diagnoses and subsequently improving patient outcomes. Citation Format: Peter A. Pellionisz, Cheng Harrison, Zachary D. Taylor, Warren Grundfest, Maie A. St. John. Dynamic Optical Contrast Imaging (DOCI): Oral screening in the underserved [abstract]. In: Proceedings of the Tenth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2017 Sep 25-28; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2018;27(7 Suppl):Abstract nr C83.

Detection of surgical margins in oral cavity cancer: the role of dynamic optical contrast imaging.

The quantity of tissue removed during an oncologic surgical procedure is not standardized and there are numerous reports of local recurrence despite histologically adequate resection margins. The oral cavity is one of the sites in the head and neck with high chances of recurrence following negative margins. To address this need, this article reviews the recent applications of Dynamic Optical Contrast Imaging (DOCI) towards both oral screening and the intraoperative evaluation of tumor margins in head and neck surgery. Human ex-vivo and in-vivo trials suggest DOCI is well tolerated, low-cost, and sensitive for differentiating cancerous from normal tissues throughout the head and neck, in addition to the oral cavity. Ex-vivo imaging of OSCC specimens generated histologically verified image contrast. Furthermore, in-vivo intraoperative results demonstrate significant potential for image-guided detection and resection of oral cavity squamous cell carcinoma (OSCC). DOCI augments tissue contrast and may enable surgeons to clinically screen patients for oral cancer, make histologic evaluations in vivo with fewer unnecessary biopsies, delineate clinical margins for tumor resection, provide guidance in the choice of biopsy sites, and preserve healthy tissue to increase the postoperative functionality and quality of life of the patient.

Dynamic Optical Contrast Imaging.

The variable location and indistinct features of parathyroid glands can make their intraoperative identification challenging. Currently, there exists no routine use of localization methods during surgery. Dynamic optical contrast imaging (DOCI) leverages a novel realization of temporally dependent measurements of tissue autofluorescence that allows the acquisition of specific tissue properties. A prospective series of patients with primary hyperparathyroidism was examined. Parathyroid lesions and surrounding tissues were collected; fluorescence decay images were acquired via DOCI. Ex vivo samples (81 patients) were processed for histologic assessment. DOCI extracts relative fluorescence decay information in a surgically relevant field of view with a clinically accessible acquisition time <2 minutes. Analysis of DOCI revealed microscopic characterization sufficient for tissue type identification consistent with histology ( P < .05). DOCI is capable of efficiently distinguishing parathyroid tissue from adjacent tissues. Such an intraoperative tool would be transformative, helping surgeons to identify lesions, preserve healthy tissue, and improve patient outcomes.

Dynamic optical contrast imaging as a novel modality for rapidly distinguishing head and neck squamous cell carcinoma from surrounding normal tissue.

Head and neck squamous cell carcinomas (HNSCCs) are debilitating diseases for which a patient's prognosis depends heavily on complete tumor resection. Currently, the surgeon's fingers determine the location of tissue margins. This study evaluated the diagnostic utility of a novel imaging modality, dynamic optical contrast imaging (DOCI), in the detection of HNSCC. This system generates contrast by illuminating the tissue with pulsed light and detecting variations in endogenous fluorophore lifetimes. A total of 47 fresh ex vivo samples from 15 patients were imaged with the DOCI system immediately after surgical resection. DOCI maps were analyzed to determine the statistical significance of contrast between tumors and adjacent nonmalignant tissue. Pilot intraoperative clinical data were also acquired. Statistical significance (P < .05) between muscle and tumor was established for 10 of 10 emission wavelengths, between collagen and tumor for 8 of 10 emission wavelengths, and between fat and tumor for 2 of 10 wavelengths. The system extracted relative fluorescence decay information in a surgically relevant field of view in <2 minutes. This study demonstrates the feasibility of using DOCI to rapidly and accurately distinguish HNSCC from surrounding normal tissue. An analysis of DOCI images revealed microscopic characterization sufficient for tissue-type identification consistent with histology. Such an intraoperative tool would be transformative by allowing the rapid delineation of tumor tissue from nontumor tissue and thus maximizing the efficacy of resection and improving patient outcomes. Cancer 2017;123:879-86. © 2016 American Cancer Society.

Dynamic Optical Contrast Imaging as a Novel Modality to Rapidly Distinguish Oral Squamous Cell Carcinoma From Surrounding Normal Tissue

Dynamic Optical Contrast Imaging as a Novel Modality to Rapidly Distinguish Oral Squamous Cell Carcinoma From Surrounding Normal Tissue