Epithelial Production, Analysis, and Innovation Core

The EPAIC prepares and distributes primary well differentiated human bronchial epithelial cultures to C3 investigators, many of whom are studying the effects of infectious agents and the response of the epithelium to these agents. The EPAIC also conducts quantitative analysis of CFTR channel activity and expression, changes in the epithelial barrier and mucociliary clearance, and multiplex analysis of cytokine and chemokine expression/secretion. The EPAIC also provides sample preparation and consulting related to ‘omics’ analysis including bulk and single cell RNAseq, proteomics, and gene editing in primary basal cells.

Immune Core

IC supports projects that seek to understand complex CF host-pathogen interactions and inflammation and thereby develop novel therapeutics that prevent the establishment of chronic infections and tissue destruction.​

Translational and Data Core

TDC was developed to supply C3 researchers with patient specimens and related clinical data to support investigation. 

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Epithelial Production, Analysis, and Innovation

​ Core

Susan Reynolds, PhD
Co-Director

Estelle Cormet-Boyaka, PhD
Co-Director

Mark Peeples, PhD
​Co-Investigator

​Our EPAIC evolved from our CFF-funded “Lung Cell Facility” which has procured and supplied CF lung explants to UNC, Chapel Hill, for the past 10 years. The UNC CF Center isolates airway epithelial progenitor cells and differentiates them into primary human bronchial epithelial (HBE) cultures. HBE cultures closely model the in vivo airway epithelium and are a critical tool for CF research. The UNC staff trained our staff to produce HBE cultures and we used the CFF funds to isolate airway progenitor cells from non-CF donor lungs, differentiate them, and supply HBE cultures to a number of NCH or OSU scientists. Recent advances in the efficiency of progenitor cell expansion, speed of maturation, and level of ciliation (up to 90%) in HBE cultures have greatly enhanced the quality of these cultures. With additional funding from CFF, our EPAIC has been able to provide more HBE cultures to more investigators and to use a small portion of the airways from explanted CF lungs to also produce CF-HBE cultures. More recently, we have begun to provide differentiated nasal epithelial (HNE) cultures from both CF and non-CF patients. We also have the ability to measure multiple chemokines and cytokines in the apical and basolateral secretions. In addition, we provide services and expertise related to ion channel expression and function as well as the assessment of mucociliary function.

Measurements Include:

1. CFTR and ENaC ion channel function using Ussing chambers
   
2. CFTR channel expression using immunoblotting and immunofluorescence
   
3. Mucociliary clearance function including airway surface liquid (ASL) height and composition and ciliary beat frequency (CBF)
   
4. Mucociliary transport (particle tracking)

HBE cultures are provided to investigators with a product sheet describing the sex, age, CFTR genotype and smoking history of the donor, and the CFTR and ENaC ion channel functions of cultures from that donor. We are developing methods to quantify cell types in HBE cultures and will also include that information with the cultures. We have also developed methods to initiate and sustain bacterial colonization of HBE cultures.

Immune Core

Santiago Partida-Sanchez, PhD
Co-Director

Amal Amer, MD, PhD
Co-Director

CF patients are prone to infections by several pathogens including Pseudomonas aeruginosa, Staphylococcus aureus, Haemophilus influenzae, Burkholderia cenocepacia, and non-tuberculosis Mycobacteria. CF patients also tend to exert hyper-inflammatory responses, which are associated with tissue damage. Therefore, it is now recognized that CF immune cells such as macrophages, neutrophils, and T cells are intrinsically dysfunctional. Our services will support projects and C3 RDP members who seek to understand complex CF host-pathogen interactions and inflammation and thereby develop novel therapeutic strategies that prevent the establishment of chronic infections and tissue destruction.

Services Include:

1. Isolation and analysis of fresh blood immune cells [neutrophils, lymphocytes, peripheral blood monocyte-derived macrophages (MDMs)], fresh bronchoalveolar lavage (BAL)-derived immune cells [alveolar macrophages (AMs), neutrophils, lymphocytes] and fresh sputum-derived immune cells (macrophages & neutrophils);
2. Critical assays of immune function including comprehensive cell counts, cytokine/chemokine production, and Enzyme-Linked ImmunoSpot (ELISpot) assays from immune cells at baseline and in response to infections or treatments;
3. Functional analysis including halide efflux and patch-clamp electrophysiology for CFTR and other ion channels within immune cells;
4. Provides expertise and training in use of the Agilent Seahorse for analysis of oxygen rate and proton efflux rate of live immune cells; and
5. Access and assistance with interpretation of state of the art ‘omics assays (single-cell RNA-Seq, mi-RNA-Seq, Methyl-Seq, CyTOF, metabolomics, proteomics) in immune cells.

Translational and Data Core

Karen McCoy, MD
​Co-Director

Nilsa Ramirez, PhD
Co-Director

​Research is integral to understanding disease and developing potential treatments. The C3 Translational and Data Core seeks to aid researchers by establishing a conduit for translational research study design and simultaneous access to patient-derived materials that will be obtained, linked with patient data, and de-identified for distribution to the cores outlined above and to individual CF researchers upon request.  We also maintain a bank of targeted biospecimens (BAL, sputum, bacterial strains) to which our researchers have easy access. Some of the biospecimens will be received as residual specimens from Projects, and others will be processed directly from CF patients as part of normal clinical care and from normal healthy controls.  While a small number of specimens will be processed for long-term storage until distribution for research, the majority of specimens will be in response to requests from investigators.