# Project 1 - Spleen

> **NIH NIH U54** · UNIVERSITY OF FLORIDA · 2020 · $580,212

## Abstract

The spleen has been considered an enigmatic and mysterious organ since its original discovery in the ancient
era. The spleen specifically controls phagocytic removal of abnormal red blood cells (RBCs) and particulate
matter, iron storage from RBCs, initial immune response to circulating antigens, and fetal hematopoiesis. Blood
filtration is anatomically associated with the red pulp whilst detection of infectious particles and initiation of an
immune response occurs via formation of germinal centers in the white pulp. The microanatomy of the human
spleen, however, has been primarily extrapolated from extensive studies in rodents. Recent studies by Dr.
Birte Steiniger (University of Marburg, Germany), have in fact indicated that as one example, the well-
delineated B-cell compartment, known as the marginal zone, between white and red pulp is distinctly absent in
the human spleen. The generation of a three dimensional (3D) tissue map of normal human spleen is therefore
timely from both the scientific and pathologic perspectives. One of our obvious strengths is extensive
experience with procurement and handling of transplant quality human organs using high level quality control
measures (AIM I). Our proposed AIMs are designed to first obtain an understanding of the overall uniformity
and macro-anatomy of the human spleen using MRI (AIM IIA). We have then designed interactive tissue
handling (formalin-fixed paraffin embedded [FFPE], optimal cutting temperature [OCT] compound embedded,
tissue clearing, and expansion) and optical microscopy (stochastic optical reconstruction microscopy [STORM],
confocal, multiphoton, light sheet fluorescence microscopy [LSFM]) pipelines, which will resolve the
microanatomy of the spleen from nanometer to millimeter in resolution (AIM IIB-D), superimposed with known
spleen biomarkers. Isolated/dispersed splenocytes will be compared to peripheral blood mononuclear cells
(PBMCs) from the same patient's blood (AIM III), and isolated immune cell subsets will be subjected to RNA-
Seq (AIM IVB). Imaging mass cytometry (IMC) will provide the ultimate co-registration of
biomolecules/biomarkers to individual cells (AIM IVA). Similarly, we will use multiplexed small molecule
fluorescence in situ hybridization (FISH) and ultimately high throughput multiplexed error-robust FISH
(MERFISH) to co-register mRNA expression to the cognate cell types (AIM IVB). Data from RNA-Seq of
individual cells will further provide new splenic biomarkers to feed back to AIMs IIB-D and IVA. The ability to
reconstruct an overall 3D spleen tissue map from all proposed pipelines is based on the common file format
used for all optical microscopy, IMC and FISH applications as delineated in the Data Core. With all of these
individual and cooperative strengths, we are poised to complete a 3D tissue map of the normal human spleen
that can be shared with all HuBMAP, HIVE and TMC investigators and ultimately, the entire scientific
community.

## Key facts

- **NIH application ID:** 9970195
- **Project number:** 5U54AI142766-03
- **Recipient organization:** UNIVERSITY OF FLORIDA
- **Principal Investigator:** MARK A. ATKINSON
- **Activity code:** U54 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $580,212
- **Award type:** 5
- **Project period:** 2018-09-14 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9970195

## Citation

> US National Institutes of Health, RePORTER application 9970195, Project 1 - Spleen (5U54AI142766-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9970195. Licensed CC0.

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