The Application Of Extracorporeal Photochemotherapy To Head And Neck Squamous Cell Carcinoma

Yale University

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Yale Medicine Thesis Digital Library School of Medicine

January 2019

The Application Of Extracorporeal

Photochemotherapy To Head And Neck

Squamous Cell Carcinoma

Alp Yurter

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Recommended Citation

Yurter, Alp, "The Application Of Extracorporeal Photochemotherapy To Head And Neck Squamous Cell Carcinoma" (2019). Yale

Medicine Thesis Digital Library. 3544.

https://elischolar.library.yale.edu/ymtdl/3544

The Application of Extracorporeal Photochemotherapy to

Head and Neck Squamous Cell Carcinoma

A Thesis Submitted to the

Yale University School of Medicine

in Partial Fulfillment of the Requirements for the

Degree of Doctor of Medicine

by

Alp Yurter

Graduating Class of 2019

TABLE OF CONTENTS

INTRODUCTION .......................................................................................................................................1

ECP Discovery ..........................................................................................................................................1

ECP’s Mechanism of Action.....................................................................................................................1

ECP’s Evolution........................................................................................................................................4

Potential Application to Head and Neck Squamous Cell Carcinoma .......................................................5

STATEMENT OF PURPOSE....................................................................................................................8

Specific Aims ............................................................................................................................................8

Hypothesis.................................................................................................................................................8

MATERIALS AND METHODS ................................................................................................................9

HPV16 E7 Antigen Sources......................................................................................................................9

Peripheral Blood Mononuclear Cells (PBMCs)......................................................................................10

CD8 T Cells.............................................................................................................................................10

Transimmunization (TI) procedure .........................................................................................................13

Cell Stimulation Readouts.......................................................................................................................15

Statistics...................................................................................................................................................16

RESULTS ...................................................................................................................................................18

REP generates a large population of CD8 T cells with Desired TCR specificity. ..................................18

CD8 T cells release IFNg upon direct stimulation with SP. ...................................................................18

Co-culture of PBMC with E7(11-20) CD8 T cells and E7 Ags results in non-specific IFNg production. .18

Co-culture of PBMC with E7(11-19) CD8 T cells and E7 Ags results in Ag-specific IFNg production. ..19

TI, platelets, and E7 Antigen sources induce a pro-inflammatory MoDC phenotype. ...........................21

PD-1 can be used as surrogate for T cell stimulation..............................................................................22

DISCUSSION.............................................................................................................................................23

Limitations and Future Directions...........................................................................................................26

REFERENCES ..........................................................................................................................................27

FIGURES....................................................................................................................................................31

ABSTRACT

Extracorporeal Photochemotherapy (ECP) is an FDA-approved immunotherapy that has been treating

cutaneous T cell lymphoma (CTCL) for over three decades. ECP’s antitumoral effect is a consequence of

its generation of functional, physiologic, inflammatory monocyte-derived dendritic cells (MoDCs) and

apoptotic, patient-derived tumor, which collectively, stimulate the adaptive immune system. Thus, in

CTCL, ECP serves as a therapeutic dendritic cell vaccine against patient-specific neoantigens. This

mechanism of action suggests ECP’s potential application to other solid tumors. We tested ECP’s

applicability to head and neck squamous cell carcinoma (HNSCC) using a trackable antigen system

involving the constitutively expressed HPV16 E7 oncoprotein. We hypothesized that ECP would

successfully stimulate anti-epitope CD8 T cells, quantified by IFN-gamma ELISA, following processing

and cross-presentation of HPV16 E7+ peptides and tumor cells by MoDCs. The trackable antigen system

employed a commonly cited epitope, E7(11-19). E7+ short peptide and long peptide generated significant

IFNg (p<0.0001) relative to the null control group. Tumor cell line SCC61 T+ (E7hi) demonstrated

significantly elevated IFNg production relative to SCC61 T- (non-E7 expressing tumor), but only in the

presence of platelets, plate-passage, and overnight incubation (p<0.0001). These results suggest an

antigen-specific CD8 T cell response and reiterate critical ECP components that have previously been

shown to facilitate immunogenic MoDC generation. Immunogenic MoDC phenotype was confirmed with

flow cytometry of inflammatory surface markers and intracellular cytokines, all of which were generally

upregulated following ECP. Overall, we have demonstrated a proof-of-principle for ECP’s therapeutic

vaccination against HNSCC. This is particularly relevant because ECP offers unique synergistic potential

with recently FDA-approved checkpoint inhibitors.