Lysosomal Biogenesis and Implications for Hydroxychloroquine Disposition

Collins, Keagan P.; Witta, Sandra; Coy, Jonathan; Pang, Yi; Gustafson, Daniel L.

doi:10.1124/jpet.120.000309

Cited by 15 publications

(17 citation statements)

References 35 publications

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“…autophagy dependent MDA-MB-231 and MDA-MB-468 tumors compared to autophagy independent MCF7 tumors (Maycotte et al, 2014) (Figure 4A), suggesting that autophagy dependent tumors sequester more HCQ over time. This could be due to an increase in lysosomes in autophagy dependent tumors compared to autophagy independent tumors (Collins et al, 2021). There are important implications of this when considering dual treatment with HCQ and other drugs, especially chemotherapies that sequester in lysosomes.…”

Section: Discussionmentioning

confidence: 99%

“…Given that AUC0-inf are not dose proportional from 40 to 80 mg/kg and clearance is concurrently increasing, this signifies non-linear PK with less drug in the blood at higher HCQ concentrations. A potential explanation is an increase in cellular lysosomal volume associated with lysosomal biogenesis triggered by HCQ (Collins et al, 2021) leading to an increase in tissue uptake and subsequent lower blood levels. This explanation is supported by the relationship between dose and AUC for blood vs. tissues where whole blood shows a 1.80 and 3.07fold increase in AUC when doses are increased 2 and 4-fold whereas tissues (brain, gut, kidney and liver) all show fold increases greater than 2 and 4 with averages of 2.47± 0.28 and 4.78 ± 0.71.…”

Section: Hcq Exposure Is Dose Dependent In Vivomentioning

confidence: 99%

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Pharmacokinetic and Pharmacodynamic Assessment of Hydroxychloroquine in Breast Cancer

Eaton

Gustafson

2021

J Pharmacol Exp Ther

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Hydroxychloroquine (HCQ) is being tested in a number of human clinical trials to determine the role of autophagy in response to standard anticancer therapies. However, HCQ pharmacodynamic responses are difficult to assess in patients and preclinical studies in mouse models are equivocal with regard to HCQ exposure and inhibition of autophagy. Here, pharmacokinetic (PK) assessment of HCQ in non-tumor bearing mice following intraperitoneal (IP) dosing established 60 mg/kg as the human equivalent dose of HCQ in mice. Autophagy inhibition, cell proliferation, and cell death were assessed in 2D cell culture and 3D tumor organoids in breast cancer. Mice challenged with breast cancer xenografts were then treated with 60 mg/kg HCQ via IP dosing and subsequent PK and pharmacodynamic (PD) responses were assessed. Although autophagic flux was significantly inhibited in cells irrespective of autophagy dependency status, autophagy dependent tumors had decreased cell proliferation and increased cell death at earlier time points compared to autophagy independent tumors. Overall, this study shows that 2D cell culture, 3D tumor organoids, and in vivo studies produce similar results and in vitro studies can be used as surrogates to recapitulate in vivo antitumor responses of HCQ.

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Section: Discussionmentioning

confidence: 99%

Section: Hcq Exposure Is Dose Dependent In Vivomentioning

confidence: 99%

Pharmacokinetic and Pharmacodynamic Assessment of Hydroxychloroquine in Breast Cancer

Eaton

Gustafson

2021

J Pharmacol Exp Ther

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“…Cell lines such as Vero (ATCC® CRL-1586™)—a monkey kidney epithelial cell line with a cell diameter of 17 μm—are usually cultured in a two-dimensional environment by completely soaking the cells in a cell culture medium containing the compound of interest. To simulate the kinetics of compound uptake, we assumed the Vero cell lysosomal volume to be 1% of its cellular volume and in the same order of magnitude as that in other cell lines [ 25 , 49 ]. The extracellular CQ and HCQ concentrations in the Vero cell line simulations were set to be constant, because bulk concentration changes were assumed to be minimal as well as in order to eliminate the dependence on differences in intracellular lysosomal volume (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…4A and D ). For example, in cells lines, the lysosomal volumes are 0.5% of the cell volume in MCF7, 0.8% in MDA-MB-231, 1.4% in T47D, and 3.7% in MDA-MB-468 cells [ 49 ]. The human respiratory tract is lined with different cell types—such as goblet cells, club cells, fibroblasts, epithelial, and macrophages—and their expression varies in different regions of the lungs.…”

Section: Discussionmentioning

confidence: 99%

Translational Modeling of Chloroquine and Hydroxychloroquine Dosimetry in Human Airways for Treating Viral Respiratory Infections

2022

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Purpose Chloroquine and hydroxychloroquine are effective against respiratory viruses in vitro. However, they lack antiviral efficacy upon oral administration. Translation of in vitro to in vivo exposure is necessary for understanding the disconnect between the two to develop effective therapeutic strategies. Methods We employed an in vitro ion-trapping kinetic model to predict the changes in the cytosolic and lysosomal concentrations of chloroquine and hydroxychloroquine in cell lines and primary human airway cultures. A physiologically based pharmacokinetic model with detailed respiratory physiology was used to predict regional airway exposure and optimize dosing regimens. Results At their reported in vitro effective concentrations in cell lines, chloroquine and hydroxychloroquine cause a significant increase in their cytosolic and lysosomal concentrations by altering the lysosomal pH. Higher concentrations of the compounds are required to achieve similar levels of cytosolic and lysosomal changes in primary human airway cells in vitro. The predicted cellular and lysosomal concentrations in the respiratory tract for in vivo oral doses are lower than the in vitro effective levels. Pulmonary administration of aerosolized chloroquine or hydroxychloroquine is predicted to achieve high bound in vitro-effective concentrations in the respiratory tract, with low systemic exposure. Achieving effective cytosolic concentrations for activating immunomodulatory effects and adequate lysosomal levels for inhibiting viral replication could be key drivers for treating viral respiratory infections. Conclusion Our analysis provides a framework for extrapolating in vitro effective concentrations of chloroquine and hydroxychloroquine to in vivo dosing regimens for treating viral respiratory infections. Graphical abstract

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“…Interestingly, the anti-infective, immunosuppressive agent hydroxychloroquine, sometimes used to counter treatment induced rashes, is reported to have lysosomotropic activity and thus might reduce treatment efficacy when co-administered with any basic drug, including CDK4/6 inhibitors. 60 Recent studies of lysosomotropism-mediated doxorubicin resistance in angiosarcoma cells suggest that this potential resistance mechanism can be countered by treating with the beta adrenergic receptor (beta-AR) antagonist propranolol, which acts through a beta-AR-independent mechanism to increase cytoplasmic doxorubicin concentrations and decrease lysosomal accumulation. 61…”

Section: Discussionmentioning

confidence: 99%

An Omic and Multidimensional Spatial Atlas from Serial Biopsies of an Evolving Metastatic Breast Cancer

Johnson

Creason

Stommel

et al. 2020

Preprint

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SummaryMetastatic cancers often respond to treatment initially but almost universally escape therapeutic control through molecular mechanisms intrinsic to tumor cells, as well as extrinsic influences from immune cells, stroma, and structural microenvironments. We explore the extent to which we can learn these mechanisms and associated therapeutic vulnerabilities by linking comprehensive molecular and multiscale imaging analyses of tumor biopsies to detailed clinical information for a patient with metastatic breast cancer. Our analyses of three serial biopsies include DNA, RNA, and protein profiling, plus quantification of tumor microenvironment composition using multiplex immunostaining and electron microscopy. Features from these analyses are linked to treatments and treatment-response measurements, including CT and PET images, circulating tumor DNA and tumor protein levels in blood, and treatment-limiting readouts of blood counts and liver function. Importantly, the measurement modalities unique to this study complemented routine diagnostics to suggest actionable mechanisms of response and resistance for each treatment phase.

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Lysosomal Biogenesis and Implications for Hydroxychloroquine Disposition

Cited by 15 publications

References 35 publications

Pharmacokinetic and Pharmacodynamic Assessment of Hydroxychloroquine in Breast Cancer

Pharmacokinetic and Pharmacodynamic Assessment of Hydroxychloroquine in Breast Cancer

Translational Modeling of Chloroquine and Hydroxychloroquine Dosimetry in Human Airways for Treating Viral Respiratory Infections

An Omic and Multidimensional Spatial Atlas from Serial Biopsies of an Evolving Metastatic Breast Cancer

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