Analytical validation and initial clinical testing of quantitative microscopic evaluation for PD-L1 and HLA I expression on circulating tumor cells from patients with non-small cell lung cancer.

INTRODUCTION: PD-L1 expression in non-small cell lung cancer (NSCLC) predicts response to immune checkpoint blockade, however is an imperfect biomarker given tumor heterogeneity, and the antigen presentation pathway requiring other components including HLA I expression. HLA I downregulation may contribute to resistance, warranting its evaluation in attempts to guide patient selection. In addition, earlier detection of acquired resistance could prompt earlier change in treatment and prolong patient survival. Analysis of circulating tumor cells (CTCs) captures heterogeneity across multiple sites of metastases, enables detection of changes in tumor burden that precede radiographic response, and can be obtained in serial fashion. METHODS: To quantify the expression of both PD-L1 and HLA I on CTCs, we developed exclusion-based sample preparation technology, achieving high-yield with gentle magnetic movement of antibody-labeled cells through virtual barriers of surface tension. To achieve clinical-grade quantification of rare cells, we employ high quality fluorescence microscopy image acquisition and automated image analysis together termed quantitative microscopy. RESULTS: In preparation for clinical laboratory implementation, we demonstrate high precision and accuracy of these methodologies using a diverse set of control materials. Preliminary testing of CTCs isolated from patients with NSCLC demonstrate heterogeneity in PD-L1 and HLA I expression and promising clinical value in predicting PFS in response to PD-L1 targeted therapies. CONCLUSIONS: By confirming high performance, we ensure compatibility for clinical laboratory implementation and future application to better predict and detect resistance to PD-L1 targeted therapy in patients with NSCLC.

Evaluation of additive manufacturing processes in the production of oculo-palpebral prosthesis.

Background: Prosthetic restorations are made to adapt or attach missing human parts in order to restore function and appearance. Maxillofacial defects connote a greater impact on patients, since the face cannot be concealed, and all the senses of the human body are expressed in it. Therefore, in order to restore the patient's quality of life, they are the ones that require the best possible adaptation to the characteristics of the patients. Methods: For the maxillofacial prostheses to fit patients, they must be personalized for each patient. The NGO "Mais Identidade" is a multidisciplinary team that specializes in the rehabilitation of patients with maxillofacial trauma. They use digital manufacturing as a tool to manufacture personalized maxillofacial prostheses for patients. With the help of the NGO, the following research is conducted with the purpose of evaluating different methods of additive manufacturing, 3D printing, in order to select the equipment that suits the needs of the method used in the manufacture of maxillofacial prostheses. To this end, eyelid models will be manufactured in different additive manufacturing equipment, and these will be evaluated according to their economic, physical, and aesthetic characteristics.