2019Joint Usage and Research Report
Title of Research Project

Role of AKT and the lysosome regulation in the induction of lymphoma in
Sjogren’s syndrome patients

ApplicantInstitution

National Institute of Dental and Craniofacial Research,

Job title

Senior Investigatorand name
Visiting

NameJohn Chiorini

researcher
Purpose of the Research Project(approx. 250 words)

Development of the Research
Project and Results(approx.. 850 words)

A major problem facing medicine today is defining the mechanism
associated with the onset and symptoms of disease and developing
treatment for them. With the reported increase in the prevalence andincidence of autoimmune diseases this group of diseases represents amajor health concern. Sjögren’s syndrome is the second most common
autoimmune disease and is estimated to affect over 35 million individuals
worldwide. This disease is characterized by a poor quality of life due tosecretory epithelial cell dysfunction with dry mouth and dry eyes,
lymphocytic infiltration within the affected salivary gland tissue,
development of autoantibodies, and a 30 fold increased risk of developing
lymphoma. Several viruses have been associated with inducing a
Sjögren’s syndrome like condition including HTLV-1, HIV, and HCV.
Currently, no effective treatment exists for Sjögren’s syndrome and ourunderstanding of the mechanism associated with the increased risk oflymphoma is not clear. The purpose of this research project is toinvestigate the molecule changes that occur in Sjögren’s associatedlymphomas and identify target therapies for intervention.
Our approach to understand the underlying mechanism of SS has been tofirst, dissect the transcriptome of minor salivary gland (MSG) frompatients (e.g., SS and non-SS) that are routinely collected for SS clinicalevaluation. We have reported that altered expression of the cytokine bonemorphogenetic protein (BMP6) is likely associated with the mostcommon symptom of primary SS (pSS), the loss of salivary gland
function in some patients. These results have suggested a novel therapy torestore gland function in these individuals.
Through a collaboration with Professor Atsumi in the Hokkaido
medical school, we have expanded our analyzes of the transcriptomes
from a diverse group of SS patients that present with different sets ofsymptoms. We are presently examining network changes that may explain
characteristics of this disease, primarily pathways associated withendoplasmic reticulum (ER) stress and constitutive activation of theunfolded protein response (UPR). This proposal will focus on one markerof this pathway, LAMP3, which is upregulated in the MSG of pSS
patients and in our preliminary experiments can induce a change in Aktand mTOR phosphorylation suggestive of a change in cellular state (e.g.,induction of apoptosis and autophagy).
The goal of this grant proposal is to: 1) Characterize LAMP3
expression in the salivary glands of pSS and non-pSS patients andunderstand the correlation between LAMP3 expression and the patientcharacteristics, 2) study the relationship between LAMP3 expression andER stress response, and 3) role of LAMP3 in salivary gland exocytosis.
Correspondingly Dr. Noguchi’ s laboratory has discovered by means ofyeast two hybrid screening using full-length Akt as a bait and RNA
aptamer technology, a connection with critical proteins and regulatory

RNA molecules in the activation of the lysosome and autophagy. Dr
Noguchi’s lab recently identified RNA aptamer specifically recognize
PI3P, which inhibits autophagy. The collaborative work with Dr.
Noguchi’s laboratory should clarify the role of Akt and autophagy in theinduction of lymphoma in Sjogren’s syndrome patients.
In this research proposal, we will use recombinant DNA experiments aswell as animal studies.
This research is part of an international effort to understand thetranscriptome of Sjogren’s syndrome and how it results in the complex setof phenotypes associated with Sjogren’s syndrome and lymphoma. The
role of LAMP3 in lysosomal function has not been as well studied as thatof other LAMP protein and its increased expression in Sjogren’ssyndrome suggests an important role in autoimmunity and the cell deathassociated with the SS. The work by Drs. Chiorini and Noguchi provides
new insight into what this role might be as a regulator of cellular state bymodulating Akt and mTOR activity. The proposed project will expand on
this as well as explore LAMP3’s role in autoantigen presentation anddeveloping animal models of altered LAMP3 expression. It is likely thatthese new concepts will influence global research on other forms ofautoimmunity. Over the past 25 years, our understanding of thebiochemical mechanisms underlying cell death has expanded greatly.
However, the connections between the signal for cell death, survival, andthe initiation of disease are not clear. LAMP3 and lysosomal function has
not been previously understood as a possible key regulator of this activity.
The innovation in our project is to link autophagy/apoptosis and thelysosome with autoimmunity.
Publication*Enter the names of conferenceor journal and its vol. No. where
the above work was presented.

【Conference, symposium, workshop etc.】An abstract on this joint project will be presented at the upcoming
International Sjogren’s syndrome meeting
Dr. Chiorini and Prof. Noguchi both presented seminars at Chiba
University and agreed to collaborate with Prof. Nakajima on the role ofautophagy in autoimmune disease.
【Journals】
A manuscript is in revision for Science Reports ( Tanaka et al. Lamp3
induces Stall autophagy and apoptosis in Sjogren’s Syndrome Patients).
Prof Atsumi and Noguchi (Hokkaido University) and Tsutomu Tanaka(former Post Doc from Prof Noguchi’s lab , and Toshio Odani (formerPost Doc from Prof Atsumi’s lab) both from Hokkaido University areco-investigators on this project.
A review is accepted for Cell Death and Disease (Noguchi et al.
Autophagy as a modulator for the cell death machinery). This is acollaboration that came about from the visit with Prof. Nakajima at Chiba
University.
A review article was accepted for Modern Rheumatology (Odani et al.
Targeting primary Sjögren's syndrome). Odani ( former Post Doc fromProf Atsumi’s lab) is from Hokkaido University and co-investigators onthis project.
A manuscript was accepted for Biochem Biophys Res Commun. (Donia etal. Identification of RNA aptamer which specifically interacts withPtdIns(3)P)). Prof Noguchi (Hokkaido University) and Tsutomu Tanaka(former Post Doc from Prof Noguchi’s lab is from Hokkaido University
and co-investigators on this project.