# Glycan engineering via exoplasmic Golgi shuttle of glycosylation building blocks and modulators

> **NIH NIH R21** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2020 · $175,839

## Abstract

Project Summary
Glycans play key roles in all aspects of biology. While harboring untapped potential as a target for biomedical
research, the glycome is still poorly characterized with respect to composition and function. In the absence of a
molecular template for glycan assembly, genetic, enzymatic and metabolic methods for the engineering of cell
surface glycan displays have been instrumental in establishing our current understanding of the physiological
and pathophysiological functions of glycans. While powerful, the current glycan engineering tools have not yet
yielded full control over the composition and structure of glycans that can be installed on living cells. A
significant challenge in glycan engineering is the delivery of the nucleotide sugar building blocks of glycans and
various modulators of glycosylation (e.g., inhibitors of glycosylation enzymes) into the organelles, where the
glycosylation machinery of cells is localized (i.e., the Endoplasmic Reticulum and the Golgi compartment).
These chemical agents are often polar or charged, and are unable to cross the multiple cellular membranes
separating the extracellular space from the secretory compartments.
 This proposal describes the development of a method for delivery of cell-impermeable nucleotide sugars
and glycosylation inhibitors directly from the culture medium into the Golgi, bypassing the cytosolic
compartment. The new method capitalizes on the intracellular trafficking of lipids between the plasma
membrane and various cellular organelles. The proposal identifies lipids at the outer leaflet of the plasma
membrane as potential carriers to shuttle cargo into the lumen of the Golgi compartment. The proposed work
will establish 1) structure-activity relationships for lipid modifications that maximize the delivery of chemical
cargo from the cell surface into the Golgi lumen, 2) optimal bioconjugation chemistries for the loading of
nucleotide sugars and glycosyl transferase inhibitors and their release in the Golgi lumen environment, and 3)
high-payload macromolecular scaffolds for the delivery and release of glycosylation modulators into the Golgi.
 A key feature of the proposed method will be the ability to alter the composition of cell surface glycans
without relying on endogenous biosynthetic and salvage pathways for the generation of nucleotide sugars as
well as transporters required for their translocation from the cytosol into the Golgi. Therefore, the proposed
method will overcome current limits on glycan structures accessible using metabolic oligosaccharide
engineering. It is also poised to offer a general mechanism for the treatment of congenital disorders of
glycosylation caused by defects in nucleotide sugar biosynthesis and transport, an approach for addressing
various pathophysiologies associated with aberrant glycosylation, and an improved method for tuning
glycosylation profiles of biologics and tissue replacements produced in non-human organisms.

## Key facts

- **NIH application ID:** 9959446
- **Project number:** 5R21GM134315-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** Kamil Godula
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $175,839
- **Award type:** 5
- **Project period:** 2019-07-01 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9959446

## Citation

> US National Institutes of Health, RePORTER application 9959446, Glycan engineering via exoplasmic Golgi shuttle of glycosylation building blocks and modulators (5R21GM134315-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/9959446. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*