Abstract
The lack of animal models for some human diseases precludes our understanding of disease mechanisms and our ability to test prospective therapies in vivo. Generation of kidney organoids from Tuberous Sclerosis Complex (TSC) patient-derived-hiPSCs allows us to recapitulate a rare kidney tumor called angiomyolipoma (AML). Organoids derived from TSC2−/− hiPSCs but not from isogenic TSC2+/− or TSC2+/+ hiPSCs share a common transcriptional signature and a myomelanocytic cell phenotype with kidney AMLs, and develop epithelial cysts, replicating two major TSC-associated kidney lesions driven by genetic mechanisms that cannot be consistently recapitulated with transgenic mice. Transplantation of multiple TSC2−/− renal organoids into the kidneys of immunodeficient rats allows us to model AML in vivo for the study of tumor mechanisms, and to test the efficacy of rapamycin-loaded nanoparticles as an approach to rapidly ablate AMLs. Collectively, our experimental approaches represent an innovative and scalable tissue-bioengineering strategy for modeling rare kidney disease in vivo.
Highlights
The lack of animal models for some human diseases precludes our understanding of disease mechanisms and our ability to test prospective therapies in vivo
We used a set of three isogenic hiPSC lines that included a line derived from an individual carrying a heterozygous 9-bp deletion in the TSC2 locus (TSC2+/−), a second isogenic TSC2−/− hiPSC line that was generated by introducing a TALEN-engineered second deletion in the wild type (WT) TSC2 allele of the patient-derived hiPSC line[19], and lastly a TSC2+/+ hiPSC line in which the original TSC2 deletion present in the patient-derived hiPSC line was corrected using CRISPR-Cas[9] (Fig. 1a)[19]
No differences in phospho-S6 levels were observed between the TSC2+/− and TSC2+/+ hiPSC lines, a finding that was consistent with the idea that one functional TSC2 allele is sufficient to preserve mTOR activity under these conditions (Fig. 1b)
Summary
The lack of animal models for some human diseases precludes our understanding of disease mechanisms and our ability to test prospective therapies in vivo. The lack of an in vivo model represents a limitation for the elucidation of disease mechanisms, and for testing prospective therapies on a preclinical level One such disease is renal angiomyolipoma (AML), a tumor found in 80% of patients with Tuberous Sclerosis Complex (TSC). Limited success was achieved using an in vivo approach that involved low-dose doxycycline-induced DNA recombination events to ablate Tsc[1] stochastically in mice carrying a ubiquitously expressed Cre transgenic allele[18] Using this strategy, small kidney lesions with characteristics of AMLs could be detected, but the resulting number of mice carrying lesions was small, reducing the suitability of this model for either mechanistic or drug-testing studies[18]. The methodology presented here is broadly applicable for the study of other rare kidney diseases for which no in vivo experimental model currently exists
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