Abstract 4886: PEGylated recombinant hyaluronidase PH20 (PEGPH20) enhances checkpoint inhibitor efficacy in syngeneic mouse models of cancer
Authors: Rosengren Sanna , Clift Renee , Zimmerman Susan J. , Souratha Jennifer , Thompson Benjamin J. , Blouw Barbara , Li Xiaoming , Zhao Qiping , Shepard Michael , Maneval Dan C. , Thanos Christopher D. , Thompson Curtis B. 
Published: July 2016

Abstract
Hyaluronan (HA), a major extracellular matrix component in many solid tumors, has been proposed to contribute to tumor progression, and to play a complex role in T lymphocyte biology. Its depletion by intravenous PEGylated recombinant human hyaluronidase PH20 (PEGPH20) remodels the tumor stroma, reduces intratumoral pressure, decompresses tumor blood vessels, and facilitates tumor drug delivery. However, the impact of HA removal on intra-tumoral immune responses and the efficacy of immune checkpoint inhibitors is unknown. To evaluate checkpoint blockade efficacy with PEGPH20, two mouse tumor cell lines, CT26 (colon) and MH194 (pancreatic, derived from spontaneous tumors in KrasLSL-G12D/+Trp53LSL-R172H/+Cre mice) were transduced with hyaluronan synthase-3 (HAS3) to generate syngeneic HA-high tumor models. For anti-CTLA4 studies, parental CT26 and CT26/HAS3 cells were implanted peritibially in Balb/C mice. While treatment with anti-mouse-CTLA4 alone (clone 9D9) inhibited tumor growth in CT26 tumors (37%), PEGPH20 alone did not significantly inhibit tumor growth or increase anti-CTLA4 efficacy. In contrast, tumor growth of CT26/HAS3 tumors was inhibited to a greater extent by the combination of PEGPH20 and anti-CTLA4 (79%) (PEGPH20 treatment 24h prior to anti-CTLA4 treatment), compared to anti-CTLA4 alone (60%, p = 0.002) or PEGPH20 alone (43%, p = 0.0001). Furthermore, gene expression of markers associated with immune suppression, such as IL10 and FoxP3, was higher in CT26/HAS3 than in CT26 tumors; suggesting an association between HA content and immune suppression. To evaluate the effect of PEGPH20 on tumor growth inhibition by PD-1 blockade, MH194/HAS3 cells were implanted peritibially in C57BL/6 mice along with immortalized pancreatic stellate cells. Growth of MH194/HAS3 tumors was significantly inhibited (33%, p = 0.049) by anti-mouse-PD-L1 antibody (clone 10F.9G2), and the addition of PEGPH20 (24h prior to anti-PD-L1) to anti-PD-L1 further enhanced tumor growth inhibition (79%, p &lt;0.0001 to both anti-PD-L1 alone and PEGPH20 alone). Similar findings were obtained with anti-mouse-PD-1 (clone RMP1-14), where the tumor growth inhibition by anti-PD-1 (33%) was further enhanced by PEGPH20 (56%, p = 0.020 and 0.017, respectively, to anti-PD-1 alone and PEGPH20 alone). At 24h following injection, the dose of PEGPH20 used (37.5 μg/kg, the human equivalent dose) removed approximately 50% of HA from tumors as shown by immunohistochemistry and HA ELISA on tumor lysates. Finally, in separate studies, PEGPH20 enhanced labelled intratumoral anti-PD-L1 accumulation (2.6 fold, p = 0.006) in a SKOV3 ovarian xenograft model engineered to express HAS2. In conclusion, in HA-high tumors, PEGPH20 reduced HA, increased anti-PD-L1 accumulation, and significantly enhanced tumor growth inhibition induced by anti-CTLA4, anti-PD-L1, and anti-PD-1 antibodies.
Citation Format: Sanna Rosengren, Renee Clift, Susan J. Zimmerman, Jennifer Souratha, Benjamin J. Thompson, Barbara Blouw, Xiaoming Li, Qiping Zhao, Michael Shepard, Dan C. Maneval, Christopher D. Thanos, Curtis B. Thompson. PEGylated recombinant hyaluronidase PH20 (PEGPH20) enhances checkpoint inhibitor efficacy in syngeneic mouse models of cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4886.