Conditional Reprogramming for Patient-Derived Cancer Models and Next-Generation Living Biobanks.

Traditional cancer models including cell lines and animal models have limited applications in both basic and clinical cancer research. Genomics-based precision oncology only help 2-20% patients with solid cancer. Functional diagnostics and patient-derived cancer models are needed for precision cancer biology. In this review, we will summarize applications of conditional cell reprogramming (CR) in cancer research and next generation living biobanks (NGLB). Together with organoids, CR has been cited in two NCI (National Cancer Institute, USA) programs (PDMR: patient-derived cancer model repository; HCMI: human cancer model initiatives. HCMI will be distributed through ATCC). Briefly, the CR method is a simple co-culture technology with a Rho kinase inhibitor, Y-27632, in combination with fibroblast feeder cells, which allows us to rapidly expand both normal and malignant epithelial cells from diverse anatomic sites and mammalian species and does not require transfection with exogenous viral or cellular genes. Establishment of CR cells from both normal and tumor tissue is highly efficient. The robust nature of the technique is exemplified by the ability to produce 2 × 106 cells in five days from a core biopsy of tumor tissue. Normal CR cell cultures retain a normal karyotype and differentiation potential and CR cells derived from tumors retain their tumorigenic phenotype. CR also allows us to enrich cancer cells from urine (for bladder cancer), blood (for prostate cancer), and pleural effusion (for non-small cell lung carcinoma). The ability to produce inexhaustible cell populations using CR technology from small biopsies and cryopreserved specimens has the potential to transform biobanking repositories (NGLB: next-generation living biobank) and current pathology practice by enabling genetic, biochemical, metabolomic, proteomic, and biological assays, including chemosensitivity testing as a functional diagnostics tool for precision cancer medicine. We discussed analyses of patient-derived matched normal and tumor models using a case with tongue squamous cell carcinoma as an example. Last, we summarized applications in cancer research, disease modeling, drug discovery, and regenerative medicine of CR-based NGLB.

Intratumor genetic heterogeneity in squamous cell carcinoma of the oral cavity.

BACKGROUND: We sought to evaluate intratumor heterogeneity in squamous cell carcinoma of the oral cavity (OCC) and specifically determine the effect of physical separation and histologic differentiation within the same tumor. METHODS: We performed whole exome sequencing on five biopsy sites-two from well-differentiated, two from poorly differentiated regions, and one from normal parenchyma-from five primary OCC specimens. RESULTS: We found high levels of intratumor heterogeneity and, in four primary tumors, identified only 0 to 2 identical mutations in all subsites. We found that the heterogeneity inversely correlated with physical separation and that pairs of well-differentiated samples were more similar to each other than analogous poorly differentiated specimens. Only TP53 mutations, but not other purported "driver mutations" in head and neck squamous cell carcinoma, were found in multiple biopsy sites. CONCLUSION: These data highlight the challenges to characterization of the mutational landscape of OCC with single site biopsy and have implications for personalized medicine.

Assessment of a Novel Computer Algorithm for Printing a 3-Dimensional Nasal Prosthetic.

Importance: The introduction and evaluation of a novel technique to create nasal prostheses with 3-dimensional (3-D) imaging software may circumvent the need for an anaplastologist. Objectives: To describe a novel computer algorithm for the creation of a 3-D model of a nose and to evaluate the similarity of appearance of the nasal prosthesis with that of the individual's nose. Design, Setting, and Participants: A prospective pilot study with a cross-sectional survey was conducted from August 1 to October 31, 2016, at a tertiary care academic center. Five volunteers were used for creation of the nasal prostheses, and 36 survey respondents with a medical background were involved in evaluating the nasal prostheses. Exposures: A computer algorithm using a 3-D animation software (Blender; Blender Foundation) and Adobe Photoshop CS6 (Adobe Systems) were used to create a 3-D model of a nose. Photographs of 5 volunteers were processed with the computer algorithm. The model was then printed using a desktop 3-D printer. Attending physicians, residents, and medical students completed a survey and were asked to rate the similarity between the individuals' photographs and their 3-D printed nose on a Likert-type scale. Main Outcomes and Measures: The similarity between 3-D printed nasal models and photographs of the volunteers' noses based on survey data. Results: Thirty-six survey respondents evaluated 4 views for each of the 5 modeled noses (from 4 women and 1 man; mean [SD] age, 26.6 [5.7] years). The mean (SD) score for the overall similarity between the photographs and the 3-D models was 8.42 (1.34). The mean scores for each nasal comparison ranged from 7.97 to 8.62. According to the survey, respondents were able to match the correct 3-D nose to the corresponding volunteers' photographs in 171 of 175 photographs (97.7%). All surveyed clinicians indicated that they would consider using this tool to create a temporary prosthesis instead of referring to a prosthodontist. Conclusions and Relevance: This algorithm can be used to model and print a 3-D prosthesis of a human nose. The printed models closely depicted the photographs of each volunteer's nose and can potentially be used to create a temporary prosthesis to fill external nasal defects. The appropriate clinical application of this technique is yet to be determined.

Ear Scaffold Reconstruction Using Ultrasonic Aspirator for Cauliflower Ear.

Untreated auricular hematomas from ear trauma can result in an ear deformation known as cauliflower ear, secondary to fibrosis and new cartilage overgrowth. Cauliflower ear reconstruction has traditionally utilized tools such as a drill or a scalpel in order to improve auricular cosmesis. We present a case report utilizing an ultrasonic aspirator to recontour the fibrosed cartilage of a cauliflower ear. The ultrasonic aspirator has advantages over traditional tools in its ability to provide finely controlled bone removal without damage to surrounding soft tissue. The patient in this case report underwent multistage reconstruction using the ultrasonic aspirator with excellent cosmetic result and patient satisfaction.

CD40 mediates downregulation of CD32B on specific memory B cell populations in rheumatoid arthritis.

Altered B cell function is important in the pathogenesis of rheumatoid arthritis (RA). In this report, we show that patients with active RA have an increased frequency of CD32B low/neg cells in the CD27(+)IgD(-) memory B cell subset and that these changes are associated with phenotypic and functional B cell activation. Studies using PBMCs from healthy donors revealed that downregulation of CD32B on B cells is mediated by CD40-CD40L interactions and is potentiated by IL-4 and inhibited by both IL-10 and IL-21. These findings appear physiologically relevant because CD4 T cell expression of CD40L correlated with the frequency of CD32B low/neg cells in the CD27(+)IgD(-) memory B subset in patients with RA. Our data support a model in which high levels of CD40L, present on circulating T cells in patients with RA, causes B cell activation and CD32B downregulation, resulting in secondary protection of memory B cells from CD32B-mediated cell death.