Identification and local delivery of vasodilators for the reduction of ureteral contractions

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Abstract

Kidney stones and ureteral stents can cause ureteral colic and pain. By decreasing contractions in the ureter, clinically prescribed oral vasodilators may improve spontaneous stone passage rates and reduce the pain caused by ureteral stenting. We hypothesized that ureteral relaxation can be improved via the local administration of vasodilators and other smooth muscle relaxants. Here, by examining 18 candidate small molecules in an automated screening assay to determine the extent of ureteral relaxation, we show that the calcium channel blocker nifedipine and the Rho-kinase inhibitor ROCKi significantly relax human ureteral smooth muscle cells. We also show, by using ex vivo porcine ureter segments and sedated pigs that, with respect to the administration of a placebo, the local delivery of a clinically deployable formulation of the two drugs reduced ureteral contraction amplitude and frequency by 90% and 50%, respectively. Finally, we show that standard oral vasodilator therapy reduced contraction amplitude by only 50% and had a minimal effect on contraction frequency. Locally delivered ureteral relaxants therefore may improve ureter-related conditions.

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Fig. 1: Establishing an in vitro model compatible with automated screening.
Fig. 2: Automated screening and single-cell analysis of hUSMC relaxation in vitro.
Fig. 3: In vitro hUSMC relaxation of screened candidates of interest.
Fig. 4: In vitro hUSMC relaxation synergies from combination doses of lead drug candidates, nifedipine and ROCKi.
Fig. 5: Ex vivo validation using porcine ureteral segments.
Fig. 6: In vivo validation of nifedipine and ROCKi.

Data availability

The data supporting the results in this study are available within the paper and its Supplementary Information. The raw datasets generated during the study are available from the corresponding author on reasonable request.

Code availability

The CellProfiler algorithms used for image analysis and for non-parametric drug synergy algorithms are included in the Supplementary Information.

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Acknowledgements

We thank A. LaRochelle and K. Wood of CBSET, Inc. for providing discarded porcine ureters used for experimentation; F. McGovern, A. Feldman and D. Dahl of Massachusetts General Hospital Department of Urology for providing human ureteral tissue samples intraoperatively; C. Beale, L. Reyelt, C. Bogins and M. Hull of Tufts Surgical Research for providing discarded porcine ureters and assisting with in vivo surgical validation and protocol design; N. Enzer, K. Cormier and G. Ekchian of the Koch Institute for primary cell line, histopathological consultations and lubricant mixing equipment, respectively; C. Lewis of the Whitehead Institute Metabolomics Core for LCMS validation; L. Richey of Tufts Comparative Pathology Services for histopathological verification and consultation related to in vivo studies; D. Logan and M. Bray of the Broad Institute for CellProfiler assistance; N. Hawes (Nicola Hawes Design) for all figure schematics. We acknowledge funding support from M. Cima, the MIT Institute of Medical Engineering and Science Broshy Fellowship (C.X.L.) and the MIT Deshpande Center for Technological Innovation (C.X.L., M.J.C. and B.H.E.). This work was also supported in part by the Koch Institute Support (core) Grant P30-CA14051 from the National Cancer Institute and the National Institutes of Health (R35 GM122547 to A.E.C.).

Author information

C.X.L., M.J.C. and B.H.E. conceived the overall study and participated in manuscript writing. C.X.L. participated in and led all experimental work and manuscript writing. J.H.C. conducted automated screening experimentation and assay design, UV-absorbance quantification and assisted with manuscript writing. B.H.E. performed surgical in vivo validation, coordination of human tissue donors and assisted with manuscript writing. C.K.S. designed the screening automation. H.D. and C.A.W. assisted with all in vitro and in vivo data analysis. K.K. and A.E.C. assisted with CellProfiler data analysis and algorithm design. A.A.B. assisted with deriving hUSMC cultures from human donors. K.S.S. assisted with study design and manuscript editing.

Correspondence to Michael J. Cima.

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Supplementary information

Supplementary Information

Supplementary Figures.

Reporting Summary

Supplementary code

CellProfiler script.

Supplementary code

CellProfiler image-quality parameters.

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Lee, C.X., Cheah, J.H., Soule, C.K. et al. Identification and local delivery of vasodilators for the reduction of ureteral contractions. Nat Biomed Eng (2019) doi:10.1038/s41551-019-0482-4

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