Diabetes Research and Clinical Practice - Volume 120 - Supplement 1

immunosuppression followed by autologous nonmyelo-

ablative hematopoietic stem cell transplantation (AHST) in

new-onset T1DM, which resulted in exogenous insulin inde-

pendence for adequate glycemic control. However, patients

with previous diabetic ketoacidosis (DKA), which is frequently

occurred in Chinese T1DM patients, were excluded in this

study. Besides, mechanisms underlying AHST-induced recon-

stitution of immune system was not clear. Thus, in 2006, we

initiated the first trial in China to examine the therapeutic

effect and mechanisms of AHST in Chinese T1DM.

From 2006 to 2008, 13 new-onset T1DM, 10 of them with DKA,

were subjected to nonmyeloablative AHST. Eleven patients

required significantly reduced doses of insulin, accompanied

by decreased HbA1c and increased C-peptide concentrations.

Three patients achieved insulin independence for 7

–

months, two of whom presented DKA at onset. The immune

system was reconstituted after AHST as characterized by the

decreased numbers of different subsets of lymphocytes within

–

6 months, and the gradually increased numbers thereafter.

Moreover, pro-inflammatory cytokines were significantly

reduced, while the levels of IL-4, IL-10 and TGF-

remained

unchanged or increased, indicating a reconstituted anti-

inflammatory environment. In parallel,

-cell antigen-specific

humoral responses was attenuated after AHST, as proved by

the decreased levels of serum islet autoantibodies. To

further determine the effect of DKA on the efficacy of AHST

in T1DM, 28 patients (11 presented DKA) who received AHST

in our and the other Chinese center were analyzed, we

finally proved that AHST achieved a greater efficacy in patients

without DKA. In 2014, D

’

Addio et al. made a multi-center

analysis by pooling data of 65 patients together from two

Chinese centers and one Polish center. It showed that 59%

achieved insulin independence within 6 months after AHST,

32% remained insulin independent till 48 months, but 52% of

treated individuals experienced adverse effects.

Nonmyeloablative AHST represents an effective treatment for

selected T1DM patients. Potential mechanisms include elim-

ination of a pro-inflammatory immune system, generation of a

new adaptive immune system, and stimulation of the regen-

eration of islet

-cells. However, the long-term effects and

safety of AHST remains to be validated, and safer HSC-based

therapeutic options are required.

S22-4

Amelioration of type 1 diabetes using direct hepatocyte

reprogramming approaches

Chia-Ning SHEN

1,4

, Fang-Pei CHANG

1,2

, Chia-Rui SHEN

Huey-Kang SYTWU

Genomics Research Center, Academia Sinica,

Graduate Institute of Life Sciences, National Defense Medical Center,

Department of Medical Biotechnology and Laboratory Science,

Chang-Gung University, Taoyuan,

Institute of Biotechnology in

Medicine, National Yang-Ming University, Taipei, Taiwan

Type 1 diabetes mellitus is characterized by complete loss of

-cells due to T-cell mediated autoimmune attack leading to a

deficiency of insulin. Transplantation of islets isolated from

the cadaveric donor pancreas has proved to be an effective

treatment for patients with type 1 diabetes mellitus. However,

this transplantation therapy is severely limited by the lack

of islet donors. Moreover, patients received the allogeneic

islet transplantationwere still suffering fromside effects of the

immunosuppressive medications. Hence, the possibility of

producing immune-tolerable

-cells would be a key challenge

for developing cell-based therapeutics for type 1 diabetic

patients. We and others had previously demonstrated hepa-

tocytes can be transdifferentiated to insulin-producing

-cells

after introducing Pdx1. Our recent findings further revealed

PDGF facilitates direct lineage reprogramming of hepatocytes

to functional

-like cells induced by Pdx1 and Ngn3. However,

it is unclear whether these transdifferentiated

-cells are

glucose responsive and immune-tolerable in autoimmune

diabetic status.

The liver not only has remarkable capacities to regenerate after

injury, but also an immune privileged organ. We therefore

hypothesize

-cells derived from hepatocyte reprograming

may possess characteristics for avoiding attack from auto-

reactive immune cells. In current work, we performed direct

conversion approaches by introducing three transcriptional

factors to primary hepatocytes isolated from Non-obese dia-

betic (NOD) mice which spontaneously develop autoimmune

diabetes. We demonstrated that simultaneously expressing

Pdx1, Ngn3, and PDGFR

could induce direct reprogramming of

hepatocytes of NOD mice to insulin-producing cells display

characteristics of pancreatic

-cells including expression of

mafa

nkx2.2

rfx6

kir6.2

glut2

and

proprotein convertase 1/3

and

possessing the capability to secrete insulin responding to

stimulatory levels of glucose. Autologous transplantation

of hepatocyte-derived

-like cells to diabetic NOD mice

significantly improved hyperglycemic status without needs

of tolerogenic treatments. Further characterization demon-

strated that

-like cells derived from hepatocyte reprogram-

ming displayed reduced levels of MHC class I molecules and

autoantigens such as

gad65

and

iapp

and also expressed PD-L1

and PD-L2. The results explain why hepatocyte-derived

-like

cells were immune-tolerable. The findings from the present

work raises the possibility of developing cell therapeutic stra-

tegy for patients with type 1 diabetes via autologous hepato-

cyte reprogramming.

Islet Imaging

S26-1

Non-invasive pancreatic beta-cell imaging using radiolabeled

exendin probe

Nobuya INAGAKI

, Hiroyuki FUJIMOTO

, Hiroyuki KIMURA

Naotaka FUJITA

, Keita HAMAMATSU

, Hideo SAJI

Department of Diabetes, Endocrinology and Nutrition, Graduate

School of Medicine, Kyoto University,

Institute for Advancement of

Clinical and Translational Science, Kyoto University Hospital,

Department of Analytical & Bioinorganic Chemistry, Kyoto

Pharmaceutical University,

Department of Patho-Functional

Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto

University, Kyoto, Japan

Impaired insulin secretion due to beta-cell dysfunction

or death as well as insulin resistance are main factors in the

pathophysiology of diabetes. Measurement of beta-cell mass

(BCM) is currently possible only by histological examination

using pancreas tissue specimens and no non-invasive method

is available for determining BCM

in vivo

. Accurate measure-

ment of BCM could provide important information on the

pathophysiology of the disease and lead to novel classifica-

tions and treatment. We have been trying to develop a non-

invasive probe to enable evaluation of BCM using radiolabeled

exendin targeting glucagon like peptide-1 receptors (GLP-1R)

expressed on beta-cells.

In this this study, we investigated whether the BCM can be

quantified non-invasively by positron emission tomography

(PET) or single-photon emission computed tomography

(SPECT) targeting GLP-1R. We synthesized [

F]-labeled, [

123

I]-

labeled, and [

111

In]-labeled exendin for PET and SPECT

imaging. These probes were evaluated by binding affinity

assay, autoradiography, biodistribution, and PET or SPECT

imaging. The probes were accumulated in murine islets and

bound to GLP-1R specifically. PET or SPECT imaging of mice

injected with these probes revealed remarkable accumulation

of radioactivity in pancreas. We then used diabetic model

mice such as streptozotosin-injected mice and NOD mice to

Speech Abstracts / Diabetes Research and Clinical Practice 120S1 (2016) S1

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