Revealing membrane alteration in cells overexpressing CA IX and EGFR by Surface-Enhanced Raman Scattering

Rusciano, Giulia; Sasso, Emanuele; Capaccio, Angela; Zambrano, Nicola; Sasso, Antonio

doi:10.1038/s41598-018-37997-3

Cited by 12 publications

(12 citation statements)

References 27 publications

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“…FACS analysis was performed according to the procedure previously described [ 89 ]. Briefly, 1 × 10 6 OVCAR3 cells were detached by using PBS EDTA 5 mM and washed 2 times with PBS.…”

Section: Methodsmentioning

confidence: 99%

Generation of a Novel Mesothelin-Targeted Oncolytic Herpes Virus and Implemented Strategies for Manufacturing

Froechlich

Gentile

Infante

et al. 2021

IJMS

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Background: HER2-based retargeted viruses are in advanced phases of preclinical development of breast cancer models. Mesothelin (MSLN) is a cell-surface tumor antigen expressed in different subtypes of breast and non-breast cancer. Its recent identification as a marker of some triple-negative breast tumors renders it an attractive target, presently investigated in clinical trials employing antibody drug conjugates and CAR-T cells. The availability of MSLN-retargeted oncolytic viruses may complement the current immunotherapeutic panel of biological drugs against HER2-negative breast and non-breast tumors. Methods: A fully virulent, tumor-targeted oncolytic Herpes simplex virus-1 (MSLN-THV) with a selectivity for mesothelin-expressing cancer cells was generated. Recombineering technology was used to replace an essential moiety of the viral glycoprotein D with antibody fragments derived from clinically validated MSLN monoclonal antibodies, and to allow IL12 cargo expression in infected cells. Panels of breast and female reproductive system cell lines were used to verify the oncolytic potential of the viral constructs. A platform for production of the retargeted viruses was developed in HEK 293 cells, providing stable expression of a suitable chimeric receptor. Results: We demonstrated the selectivity of viral infection and cytotoxicity by MSLN-retargeted viruses in a panel of mesothelin-positive cancer cells, originating from breast and female reproductive system tumors. We also developed a second-generation oncolytic MSLN-THV, encoding IL12, to enhance the immunotherapeutic potential of the viral backbone. A non-tumor cell line expressing a chimeric MSLN/Nectin-1 receptor, de-sensitized from antiviral responses by genetic inactivation of the Stimulator of Interferon Genes (STING)-dependent pathway was engineered, to optimize viral yields. Conclusions: Our proof-of-concept study proposes MSLN-retargeted herpesviruses as potential cancer immunotherapeutics for assessments in preclinical models of MSLN-positive tumors, complementing the available panel of oncolytic viruses to HER2-negative breast tumors.

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“…FACS analysis was performed according to the procedure previously described [ 89 ]. Briefly, 1 × 10 6 OVCAR3 cells were detached by using PBS EDTA 5 mM and washed 2 times with PBS.…”

Section: Methodsmentioning

confidence: 99%

Generation of a Novel Mesothelin-Targeted Oncolytic Herpes Virus and Implemented Strategies for Manufacturing

Froechlich

Gentile

Infante

et al. 2021

IJMS

Self Cite

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“…Negative selection steps can, of course, remove, at least in part, these non-specific phages. In the case of an antibody phage display screening performed on living cells, it is quite hard to identify an ideal substrate for negative selection, unless two background-matched, isogenic cell lines differing only in target expression are available (e.g., use of target overexpression in a target-negative cell line or knock out cell clones) [ 39 , 76 , 77 , 78 ]. Removal of phages reactive for molecular baits as well as solid-phase components is also challenging.…”

Section: Implementation Of Next Generation Sequencing To Phage Displa...mentioning

confidence: 99%

High-Throughput Monoclonal Antibody Discovery from Phage Libraries: Challenging the Current Preclinical Pipeline to Keep the Pace with the Increasing mAb Demand

Zambrano

Froechlich

Lazarevic

et al. 2022

Cancers

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Monoclonal antibodies are among the most powerful therapeutics in modern medicine. Since the approval of the first therapeutic antibody in 1986, monoclonal antibodies keep holding great expectations for application in a range of clinical indications, highlighting the need to provide timely and sustainable access to powerful screening options. However, their application in the past has been limited by time-consuming and expensive steps of discovery and production. The screening of antibody repertoires is a laborious step; however, the implementation of next-generation sequencing-guided screening of single-chain antibody fragments has now largely overcome this issue. This review provides a detailed overview of the current strategies for the identification of monoclonal antibodies from phage display-based libraries. We also discuss the challenges and the possible solutions to improve the limiting selection and screening steps, in order to keep pace with the increasing demand for monoclonal antibodies.

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“…For instance, electric cell-impedance sensing (ECIS) methods can easily reveal the formation of cell–cell junctions and cell-substrate contacts, and are extensively used to follow epithelial maturation ( Ngok et al, 2013 ; Gamal et al, 2017 ; Van Der Stoel et al, 2020 ). Surface enhanced Raman spectroscopy (SERS) is ideal to obtain information about biochemical changes inside cells in the proximity of a nanostructured surface ( El-Said et al, 2015 ; Haldavnekar et al, 2018 ; Rusciano et al, 2019 ).…”

Section: Relevance Of Biosensors In Cell Adhesion Studiesmentioning

confidence: 99%

Biosensors for Studies on Adhesion-Mediated Cellular Responses to Their Microenvironment

Saffioti

Cavalcanti‐Adam

Pallarola

2020

Front. Bioeng. Biotechnol.

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Cells interact with their microenvironment by constantly sensing mechanical and chemical cues converting them into biochemical signals. These processes allow cells to respond and adapt to changes in their environment, and are crucial for most cellular functions. Understanding the mechanism underlying this complex interplay at the cell-matrix interface is of fundamental value to decipher key biochemical and mechanical factors regulating cell fate. The combination of material science and surface chemistry aided in the creation of controllable environments to study cell mechanosensing and mechanotransduction. Biologically inspired materials tailored with specific bioactive molecules, desired physical properties and tunable topography have emerged as suitable tools to study cell behavior. Among these materials, synthetic cell interfaces with built-in sensing capabilities are highly advantageous to measure biophysical and biochemical interaction between cells and their environment. In this review, we discuss the design of micro and nanostructured biomaterials engineered not only to mimic the structure, properties, and function of the cellular microenvironment, but also to obtain quantitative information on how cells sense and probe specific adhesive cues from the extracellular domain. This type of responsive biointerfaces provides a readout of mechanics, biochemistry, and electrical activity in real time allowing observation of cellular processes with molecular specificity. Specifically designed sensors based on advanced optical and electrochemical readout are discussed. We further provide an insight into the emerging role of multifunctional micro and nanosensors to control and monitor cell functions by means of material design.

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Revealing membrane alteration in cells overexpressing CA IX and EGFR by Surface-Enhanced Raman Scattering

Cited by 12 publications

References 27 publications

Generation of a Novel Mesothelin-Targeted Oncolytic Herpes Virus and Implemented Strategies for Manufacturing

Generation of a Novel Mesothelin-Targeted Oncolytic Herpes Virus and Implemented Strategies for Manufacturing

High-Throughput Monoclonal Antibody Discovery from Phage Libraries: Challenging the Current Preclinical Pipeline to Keep the Pace with the Increasing mAb Demand

Biosensors for Studies on Adhesion-Mediated Cellular Responses to Their Microenvironment

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