Collaborative Project #8: Novel Strategies for Immunological Reprogramming 

Collaborating Investigator: Dino Di Carlo, PhD 
Affiliation: Cleveland Clinic
Director, Lerner Research Institute, Cleveland Clinic 

Funding Source: NSF
Grant Number: 2149551  
Project Period: 09/15/22-08/31/25

Significance

The Di Carlo Lab leverages microfluidics, microfabrication, and nanotechnologies to develop interfaces with cells and molecules for applications in disease diagnosis, therapeutic discovery, tissue regeneration, and directed evolution. Instead of scaling down macroscale concepts, the Di Carlo lab takes advantage of unique physics at small scales to process and analyze cells and manufacture smart biomaterials structured at sizes smaller than a human hair. They aim to engineer solutions that utilize commonly available laboratory equipment and consumer electronic devices so that they can be readily adopted by researchers, doctors, and patients. These tools not only excel at the manipulation and analysis of single cells and molecules, but also are cost-effective and easily accessible – thus democratizing advanced biotechnology capabilities to solve medical problems.

Various single-cell functional profiling platforms have emerged over the past several years, but their widespread adoption has been limited due to low assay throughput, high costs, or the need for skilled operators and expensive customized instrumentation. Our collaborators are developing broadly accessible platforms to study immune activation at single-cell resolution by isolating individual T cells into uniformly sized nano-droplets, each formed by a microparticle with a structured cavity (termed a nanovial). This technology will allow researchers to separate individual cells based on their molecular secretion profiles.

Approach

Aim:  The Di Carlo lab has developed methods to fabricate hydrogel-based chemically functionalized microcontainers, called nanovials, and demonstrated their use for sorting single viable cells based on their secreted products at high-throughput using only commonly accessible laboratory infrastructure. These nanovials act as solid supports that facilitate attachment of a variety of adherent and suspension cell types, partition uniform aqueous compartments, and capture secreted proteins. This CP will use NP-based and HMP-based aAPC to validate the nanovial platform for assessment of immune function cells.

Push-Pull relationship

We will integrate aAPC technologies from TR&D1 and immunocytokine (IC) technologies from TR&D3 into Dr. the Di Carlo lab novel “lab on a particle” nanovial platform. Analysis of aAPC and IC technologies will illustrate the utility and versatility of nanovials, while also providing insight into which cellular readouts are most important to evaluate. This will in turn enhance platform design and development. Pull: We will characterize the functional properties of individual cells stimulated by NCBIB technologies using the nanovial platform. This will inform mechanistic understanding of the deliverables developed in TR&D1 and TR&D3 and suggest alternate NP-based and HMP based aAPC.

TRD