Modeling radiation injury andcountermeasure drug responses in a humanIntestine Chip
Sasan Jalili-Firoozinezhad
11.16.18(Lab of Don Ingber, MD,PhD; Wyss Institute at Harvard University)

Organ-on-Chip Design Goals• Recreate Tissue-Tissue Interface that define Organ Structure• Provide Mechanical Cues necessary for relevant physiology• Precisely Orient Cells for High-Resolution Real-Time Imaging• Control Fluid Flow through Microfluidic Channels• Incorporate Endothelium-Lined Vascular Channels- To enable physiological vascular coupling between different organ chips- Permits real-time analysis of inflammation (recruitment of circulating immune cells)

Ingber, Development, 2018

Dose-dependent radiation sensitivities acrossdifferent human organs
Lung
2 Gy

6 Gy•
•
•
•

Edema
Cell barrier degeneration
Alveolitis/pneumonitis
Pulmonary fibrosis

Gut•
•
•
•

Villi blunting
Tight junctions disruption
Increased apoptosis
Mucosal barrier breakdown

12 Gy

Bone Marrow••••

Most radiosensitive organ
2 Gy dose clinically significant
Life-threatening cytopenias
Few effective interventions

Human Intestine-on-a-Chip
Project Goal: Determine whether Intestine Chip can serve as in vitro alternatives to in vivo
screening of Medical Countermeasures (MCMs) for Acute Radiation Syndrome.

Radiation-induced Destruction ofEpithelial Junctions is Mitigated by DMOGConRadRadRad

DMOG+RadDMOG

DMOG

ZO1
DAPI

100 mmVE-Cad
DAPI

20 mm

Jalili et al., Cell Death & Disease, 2018

DMOG Prevents Radiation-Induced Villus Blunting
and Loss of Intestinal Microvilli
Villin DAPI

100 mm

50 mm

1 mm

10 mm

Jalili et al., Cell Death & Disease, 2018

Gut endothelium mediates radiation damage- Endothelium+ Endothelium

RadConZO1
MUC2DAPI

50 mm

Jalili et al., Cell Death & Disease, 2018

Our primary human Intestine Chip may mimic invivo intestinal physiology better than Gut Chip
• Caco-2 cell line can recapitulate many features of the humanileum, however, since it is a tumor cell line it might differ insensitivity to radiation.

• This could provide us with the opportunity to evaluatepatient-specific responses to radiation as well.
DIC

100 mmMUC5AC/Factin/DAPI

MUC5AC/Factin/DAPI

100 mm

50 mm

Ki67/Factin/DAPI

50 mm

Kasendra et al., Sci Rep, 2018Jalili-Firoozinezhad et al., unpublished data

Villus blunting and reduced barrier function atlower radiation exposure
Ctrl

4 Gy

8 Gy

12 Gy

***

*Primary intestinal epithelium is more radio-sensitive compared to Caco-2
epithelium, as we did not detect any damage at 4 Gy in Caco-2 cells

4 Gy

8 Gy

12 Gy

Epithelium

Ctrl

Endothelium

50 mm

50 mm

MUC5AC, VE-Cadherin, Factin, DAPI• Reduced MUC5AC+ cells in irradiated samples• Villus blunting and loss of junctions in irradiated samples
Jalili-Firoozinezhad et al., unpublished data

Does the Gut Microbiota DetermineRadiosensitivity of Host?

Cui et al., EMBO, 2017

Cho et al., Nature, 2012

Donaldson et al., Nature, 2016•

Establish complex microbiota anaerobic co-culture in the Intestine Chip tostudy how the gut microbiome contributes to radiation response•

We have developed methods to culture complex communities of commensal
microbes that comprise the human gut microbiome in our Gut Chip

Complex Human Gut Microbiome Cultured In An
Anaerobic Human Gut-On-A-Chip

O2

Epithelium

Endothelium

1

BTBPDMSBT
150mm

TSensorEpithelium
Endothelium
Sensor

234PDMS

Jalili-Firoozinezhad et al., under review

Emulating Intestinal Villi Height Gradients On Chip
Epithelium

Aerobic

Villin DAPI

Anaerobic*Endothelium

ZO1
DAPI

CD144
DAPI

100 mm

20 mm

100 mmLongitudinal sections of small intestine of adult male rats in five locations

Wang et al., CMGH, 2018

Altmann et al., Am J Anat, 1970

Jalili-Firoozinezhad et al., under review

Co-culture of human intestinal epithelium andobligate anaerobe, Bacteroides fragilis, on-chipJalili-Firoozinezhad et al., under review

Anaerobic gut chip sustains a complex humanintestinal microbiome in vitroCo-culture of fresh human patient-derived gut
microbiome with primary human ileal Intestine Chipab
Merge

VillinFactincd
Ileum sample (day 5)

Observedrichness

Shannondiversity

36 wk infant

122

0.32

36 wk infant

118

0.34

36 wk infant

121

0.31

30 wk infant

135

1.10

Jalili-Firoozinezhad et al., under review

DAPI

Summary• The Human Gut Chip Recapitulates Acute Radiation
Injury and pre-treatment with the MCM drug, DMOG,
significantly reduces radiation-induced injury on chip.
• We developed methods to culture complex human gut
microbiome in an anaerobic human Intestine Chip.
• Future experiments will be focused on:• Using Human Intestine Chip in combination with multiomics and bioinformatics to discover new MCMs.
• Studying the role of gut microbiome in response toradiation in Human Intestine Chip.

Don Ingber

Acknowledgement

Rachelle Prantil-Baun
Elizabeth Calamari
Oren Levy
Amy WenAmanda Jiang
David Chou
Vik Frismantas
Yuka Milton
Amir Bein
Cicely Fadel
Diogo CamachoOlivier Henry
Ratnakar Potla
Bret Nestor
Francesca Gazzaniga
Alessio Tovaglieri
James Weaver
Tom Ferrante

Our Collaborators:
Dennis Kasper (HMS)David Breault (BCH)
Joaquim Cabral (IST, Portugal)