Project Summary
A broad range of inflammatory gastrointestinal conditions impact millions of people in the United States and
abroad. The etiology of the disease is multifactorial and heterogeneous, but the common underlying feature is
an overactive immune system. Genetics, environmental factors, age, sex, race and even the composition of
bacteria that reside in the gut lumen are associated with causation of gut inflammation. While some therapeutic
advances have been made, a large number of patients do not respond to existing pharmaceutical interventions
leaving many to have life-long morbidity. This provides strong rationale to improve existing therapies and
discover alternative and novel approaches to deal with inflammatory conditions of the gut. Preclinical models
used for drug research and discovery have been historically poor. They are comprised cancer cells that have low
physiological relevance and do not possess the immune cell compartment, which is a missing pivotal component
to achieve an accurate model of human inflammatory conditions. For these reasons, there is a need in the
therapeutics marketplace for an in vitro intestinal platform that accurately recapitulates luminal-epithelial-
immune cell axis of the intestines. To meet this need, Altis Biosystems Inc., an early-stage biotechnology
company, has collaborated with scientists at academic laboratories to develop a novel, primary-stem cells-based,
in vitro intestinal model (termed RepliGut). We have finished an SBIR Phase I program focused on transgenesis
and gene editing of intestinal stem cells (ISCs) cultured on RepliGut. We developed reporter-gene ISCs that
generate differentiated epithelium and sense and report in real-time key features of gut inflammation, 1) barrier
function, and 2) NF-kB activation. Here we expand the scope to generate a first-in-kind co-culture model called,
InflammaGutTM. InflammaGut is a tripartite and flexible system that uses fundamental technology of RepliGut and
superimposes inflammation reporter genes and co-culture of human epithelium with allogeneic gut-associated
lymphocytes (GAL) to recreate the LEI-axis. Four products are envisioned: 1) InflammaGut:ZO1 and
InflammaGut:NF-kB reporter gene epithelium, 2) InflammaGut:Co-Culture: an epithelial/gut-associate lymphocyte
(GAL) co-culture system, 3) commercializable human gut-associated lymphocytes, and 4) a new contract
research service (CRS) using InflammaGut. There is strong engineering and biological method innovation,
including, 1) development of Transwell insert enabling scalable culture of RepliGut differentiated human
epithelium over a hydrogel encapsulated lymphoid-cell compartment, and 2) isolation, culture, and cryobanking
of GAL from human organ donors. InflammaGut will be built, validated, and tested by academic collaborators
(founders of the RepliGut technology), Altis Biosystems, and two industry collaborators.
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