Generation and Characterization of a Breast Cancer Resistance Protein Humanized Mouse Model

Dallas, Shannon; Salphati, Laurent; Gómez-Zepeda, David; Wanek, Thomas; Chen, Liangfu; Chu, Xiaoyan; Kunta, Jeevan; Mezler, Mario; Menet, Marie-Claude; Chasseigneaux, Stéphanie; Declèves, Xavier; Langer, Oliver; Pierre, Esaie; DiLoreto, Karen; Hoft, Carolin; Laplanche, Loı̈c; Pang, Jodie; Pereira, Tony; Andonian, Clara; Simic, Damir; Rode, Anja; Yabut, Jocelyn; Zhang, Xiaolin; Scheer, Nico

doi:10.1124/mol.115.102079

Cited by 23 publications

(20 citation statements)

References 41 publications

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“…However, several lines of evidence indicate that there are species differences. To overcome this problem for modeling, mice humanized for BBB transporters have been generated and characterized recently, such as a mouse line humanized for P-glycoprotein and breast cancer resistance protein (Sadiq et al, 2015;Dallas et al, 2016). However, compared with those humanized for efflux transporters that do exist, to our knowledge, no mice humanized for uptake transporters have existed until now.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Organic Anion Transporter 1A2-knock-in Mice as a Model of Human Blood-brain Barrier

Sano

Mizuno

Mochizuki

et al. 2018

Drug Metabolism and Disposition

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The present study aimed to establish a humanized mouse model with which to explore OATP1A2-mediated transcellular transport of drug substrates across the blood-brain barrier (BBB) and to evaluate the usefulness of the humanized mice in preclinical studies. Sulpiride, amisulpride, sultopride, and triptans were used as probes to discriminate OATP1A2 and Oatp1a4. We generated a mouse line humanized for OATP1A2 by introducing the coding region downstream of the Oatp1a4 promoter using the CRISPR/Cas9 technique. In the mice generated, OATP1A2 mRNA in the brain was increased corresponding to disappearance of Oatp1a4. OATP1A2 was localized on both the luminal and abluminal sides of the BBB. Unfortunately, study in vivo employing sulpiride, sumatriptan, and zolmitriptan as probes did not indicate any difference in their brain-to-plasma ratio between the control and humanized mice. Quantitative targeted absolute proteomic analysis of the BBB fraction from the humanized mice revealed that almost all analyzed transporters and membrane proteins were expressed at similar levels to those in control mice. The quantitative levels of OATP1A2 differed depending on the peptide quantified, which suggests that incomplete translation or posttranslational modification may occur. The blood-to-brain transport of zolmitriptan, determined by brain perfusion in situ, was 1.6-fold higher in the humanized mice than in the controls, whereas that of sulpiride was not significantly changed. To our knowledge, we established a mouse line humanized for a BBB uptake transporter for the first time. Unfortunately, because of limited impact, there is still room for improvement of the model system.

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Section: Discussionmentioning

confidence: 99%

Evaluation of Organic Anion Transporter 1A2-knock-in Mice as a Model of Human Blood-brain Barrier

Sano

Mizuno

Mochizuki

et al. 2018

Drug Metabolism and Disposition

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“…Among other applications, TEP has been used to asses P-gp and BCRP function and inhibition in mouse [238,253,270], rat [261,271], primate [250,264] and human [251,254] models; and it could be used for precision medicine through the evaluation of individual variability in response to CNS drugs [272]. In addition, diverse strategies have been developed to study for instance drug-drug interactions of unlabeled molecules in combination with already available tracers [246].…”

Section: Imaging Methodsmentioning

confidence: 99%

“…In addition, diverse strategies have been developed to study for instance drug-drug interactions of unlabeled molecules in combination with already available tracers [246]. PET was recently employed to validate the permeability and transporter function of a human iPSCs BBB model [172] and the evaluation of ABC transporter-humanized mice models [238,270]. Importantly, it has been used to evaluate drug delivery strategies, such as inhibition [250,251] or focused ultrasounds [273] (see Section 6.5 for details).…”

Section: Imaging Methodsmentioning

confidence: 99%

“…It is important to remember that there can be molecular and physiological differences between species, such as expression levels, substrate specificity, and transport efficiency in the case of ABC transporters (see Section 3.6). Therefore, models with humanized ABC transporters, as P-gp and BCRP, have been developed to better represent drug transport in humans [236][237][238][239]. Diverse techniques are used to evaluate the permeability of molecules into the brain and their clearance and obtain important ADME, toxicity, and PK-PD parameters (reviewed in [5,6,8,133,240].…”

Section: In Vivo Models and Assaysmentioning

confidence: 99%

“…The results of this absolute quantification can be used to study transporter expression modulation due to diseases or drug exposure [223], interspecies [93], or even human interindividual variability [305]; besides, results have been used to calculate the relative expression factors (REF) for IVIVE [126,295] and PBPK modeling (see [306] for recommendations). In addition, the specificity of these methods has been exploited to characterize and validate murine models with humanized ABC transporters [236][237][238]270].…”

Section: Recent Molecular Characterization Techniquesmentioning

confidence: 99%

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ABC Transporters at the Blood–Brain Interfaces, Their Study Models, and Drug Delivery Implications in Gliomas

et al. 2019

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Drug delivery into the brain is regulated by the blood–brain interfaces. The blood–brain barrier (BBB), the blood–cerebrospinal fluid barrier (BCSFB), and the blood–arachnoid barrier (BAB) regulate the exchange of substances between the blood and brain parenchyma. These selective barriers present a high impermeability to most substances, with the selective transport of nutrients and transporters preventing the entry and accumulation of possibly toxic molecules, comprising many therapeutic drugs. Transporters of the ATP-binding cassette (ABC) superfamily have an important role in drug delivery, because they extrude a broad molecular diversity of xenobiotics, including several anticancer drugs, preventing their entry into the brain. Gliomas are the most common primary tumors diagnosed in adults, which are often characterized by a poor prognosis, notably in the case of high-grade gliomas. Therapeutic treatments frequently fail due to the difficulty of delivering drugs through the brain barriers, adding to diverse mechanisms developed by the cancer, including the overexpression or expression de novo of ABC transporters in tumoral cells and/or in the endothelial cells forming the blood–brain tumor barrier (BBTB). Many models have been developed to study the phenotype, molecular characteristics, and function of the blood–brain interfaces as well as to evaluate drug permeability into the brain. These include in vitro, in vivo, and in silico models, which together can help us to better understand their implication in drug resistance and to develop new therapeutics or delivery strategies to improve the treatment of pathologies of the central nervous system (CNS). In this review, we present the principal characteristics of the blood–brain interfaces; then, we focus on the ABC transporters present on them and their implication in drug delivery; next, we present some of the most important models used for the study of drug transport; finally, we summarize the implication of ABC transporters in glioma and the BBTB in drug resistance and the strategies to improve the delivery of CNS anticancer drugs.

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Breast Organotypic Cancer Models

Carranza-Rosales

Guzmán-Delgado

Carranza-Torres

et al. 2018

Current Topics in Microbiology and Immunology

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Breast cancer is the most common cancer type diagnosed in women, it represents a critical public health problem worldwide, with 1,671,149 estimated new cases and nearly 571,000 related deaths. Research on breast cancer has mainly been conducted using two-dimensional (2D) cell cultures and animal models. The usefulness of these models is reflected in the vast knowledge accumulated over the past decades. However, considering that animal models are three-dimensional (3D) in nature, the validity of the studies using 2D cell cultures has recently been questioned. Although animal models are important in cancer research, ethical questions arise about their use and usefulness as there is no clear predictivity of human disease outcome and they are very expensive and take too much time to obtain results. The poor performance or failure of most cancer drugs suggests that preclinical research on cancer has been based on an over-dependence on inadequate animal models. For these reasons, in the last few years development of alternative models has been prioritized to study human breast cancer behavior, while maintaining a 3D microenvironment, and to reduce the number of experiments conducted in animals. One way to achieve this is using organotypic cultures, which are being more frequently explored in cancer research because they mimic tissue architecture in vivo. These characteristics make organotypic cultures a valuable tool in cancer research as an alternative to replace animal models and for predicting risk assessment in humans. This chapter describes the cultures of multicellular spheroids, organoids, 3D bioreactors, and tumor slices, which are the most widely used organotypic models in breast cancer research.

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Generation and Characterization of a Breast Cancer Resistance Protein Humanized Mouse Model

Cited by 23 publications

References 41 publications

Evaluation of Organic Anion Transporter 1A2-knock-in Mice as a Model of Human Blood-brain Barrier

Evaluation of Organic Anion Transporter 1A2-knock-in Mice as a Model of Human Blood-brain Barrier

ABC Transporters at the Blood–Brain Interfaces, Their Study Models, and Drug Delivery Implications in Gliomas

Breast Organotypic Cancer Models

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