# A developmental engineering toolbox for large-scale tissue engineering

> **NIH NIH R35** · UNIVERSITY OF PENNSYLVANIA · 2020 · $176,042

## Abstract

PROJECT SUMMARY
Many diseases in complex, hierarchically organized tissues such as the breast, lung, and prostate have been
difficult to address, because they are a product of complex multicellular dynamics. For example, congenital
diseases of the kidney are staggeringly common. Around a third of all birth defects are associated with
problems in kidney and urinary tract development, but researchers have few options for capturing the full
functional complexity of this organ system outside of the body. This is because current kidney models are
either 2D, single cell-type approximations, or are organoid models with more cellular diversity, but with little of
the long-range spatial structure that is crucial for kidney function.
 The Hughes lab aims to solve two critical engineering barriers to the development of better in
vitro human tissue models. First, we aim to standardize and vastly increase the throughput of organoid-
based phenotypic screens related to human disease. Second, we aim to bring an entirely new philosophy to
tissue engineering, in which tissue scaffolds are not built in final form, but rather as immature “seeds” that are
guided through developmental transitions in structure that mimic those of their target tissue. These transitions
morph flat tissue scaffolds into final tissue forms that achieve defined shapes, cell distributions, and ECM
compaction and alignment patterns in 3D that establish a new way of building hierarchical tissues like the
kidney.
 To the first aim, we propose to re-engineer our cell DNA “velcro” cell and organoid patterning
technology. This technology allows us to precisely pattern multiple cell types with single-cell resolution at the
interface with organotypic gel layers, yet its throughput is currently limited. We will apply a photopatterning
approach in which cell-adhesive ssDNA strands can be patterned in millions of locations simultaneously, a key
requisite for whole-genome organoid screens. Secondly, we propose high-throughput pluripotent stem cell
patterning and culture technologies that reduce inter-organoid variation, to enable whole genome CRISPR-
based screening for genetic risk factors of disease, using kidney organoids as a prototypical system. To the
second aim, we build upon our recent description of dynamic tissue scaffolds to position organoids in
3D using autonomously folding gels that couple their niches through tracts of dynamically remodeled ECM.
Using these centimeter-scale, 3D organoid patterning capabilities, we envision an analogy between the
branching pattern of the kidney collecting duct network and the edge networks of “flasher” origami patterns. By
controlling the morphogenesis of these patterns, we seek to engineer the progressive formation of a
contiguous collecting duct network between locally self-organizing tissue niches. Rather than directly
building tissues in a final, yet immature form, we believe that building hierarchical tissues by guided
morphogenesis presents a transf...

## Key facts

- **NIH application ID:** 10149089
- **Project number:** 3R35GM133380-02S1
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** Alex Hughes
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $176,042
- **Award type:** 3
- **Project period:** 2019-08-01 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10149089

## Citation

> US National Institutes of Health, RePORTER application 10149089, A developmental engineering toolbox for large-scale tissue engineering (3R35GM133380-02S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10149089. Licensed CC0.

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