A Platform for High-Resolution Phenotyping of Cells for Functional Characterization, Enrichment and Targeting

Background

The ability to identify and enrich distinct cell types from a heterogeneous population is an extremely powerful method for developing disease interventions, diagnostics, and drug delivery tools. It is also a critical component of basic scientific research to dissect the mechanistic features of complex biological processes. Current cell characterization methods, such as flow cytometry, are limited by the properties of cluster-based differentiation (CD) markers, and do not reveal the extensive functional and cellular heterogeneity reported by nucleic acid-based methods (e.g., single-cell sequencing). However, nucleic acid methods destroy the cells, thereby precluding the isolation of highly purified cells for downstream applications, analyses, or the development of therapeutic interventions.

The current invention overcomes these limitations through its unprecedented resolution, maintenance of cell integrity, and orthogonality with existing cell characterization methods.

Technology Overview

Drs. Hannes Buelow, Ulrich Steidl, and Steven Almo have developed a novel method of characterizing cells at higher granularity than possible with existing technologies.

Cell populations differ in the heterogeneity of heparan sulfate (HS) glycan modifications presented on the plasma membrane, resulting in unique glycan signatures (glycotypes). Unlike the CD marker-based system, which relies on a limited set of markers, the new strategy enables a much more granular dissection of subsets within a heterogeneous population as it leverages the extraordinary molecular diversity of HS chains. A panel of anti-HS single-chain variable fragment antibodies (scFVs) has been developed that is capable of detecting glycan signatures unique to cell types ().

In proof-of-concept studies, the team successfully separated human and mouse hematopoietic progenitor cells, previously defined as a homogeneous population on the basis of existing CD markers, into distinct subpopulations of immunophenotypically pure megakaryocyte-erythrocyte progenitors with distinct proliferative properties (Piszczatowski et al. 2022). Unpublished data applying the same strategy to other cell types and tissues can be made available under CDA.

The utility of this technology has been demonstrated in the hematopoietic lineage and will be expanded to many other cell lineages and systems, enabling the discrimination of cell types in normal versus disease contexts, and across other cell types. In addition, the technology has immense potential to be further developed into a platform for precision drug delivery, diagnostics, and immunotherapeutics targeting specific cell populations. Importantly, the technology is non-destructive to the cells under study, allowing their propagation for downstream applications following isolation.

Further Details:

• Piszczatowski et al (J. Exp. Med. 2022)
• Attreed et al., Glycobiology (2016)
• Attreed et al., Nat Methods (2012)

Stage of Development

• Proof-of-concept established using human and mouse hematopoietic cell lineages
• Studies in other tissue types and (patho)physiological conditions are ongoing and can be made available under CDA[US1].

Benefits

• High resolution compared to conventional cell separation and targeting methods ()
• Easily integrated into current workflows for analysis and sorting using flow cytometry
• Ability to combine with CD-based separation as an orthogonal tool
• Not limited by knowledge of surface markers
• Non-destructive, allowing live cells to be propagated for further applications
• The library/panel size is scalable and can, in principle, be substantially expanded

Applications

• Functional characterization and enrichment of cells from heterogeneous cell populations
• Discrimination and phenotyping of cell types under healthy and disease states for use as diagnostic tools
• Targeting distinct cell populations for therapeutic intervention and drug delivery

Opportunity

• Licensing
• Collaboration
• Developmental partner