Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion

一种人工抗原、细胞的技术，应用在受体/细胞表面抗原/细胞表面决定因子、抗肿瘤药、动物细胞等方向

Inactive Publication Date: 2019-10-08
IOVANCE BIOTHERAPEUTICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

EM-3 cells and the closely related EM-2 cell line have not been previously reported as useful aAPCs for cell expansion for tumor immunotherapy applications

Method used

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  • Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion
  • Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion
  • Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0607] Example 1 - Variability in Expansion of Tumor Infiltrating Lymphocytes Using PBMC Feeder Cells

[0608] By comparing the results of multiple TIL expansions on the same TIL line obtained from a patient, the variability in TIL expansion obtained by using PBMC feeder cells can be demonstrated. figure 1 Typical results for rapid expansion of TILs using irradiated allogeneic PBMC feeder cells (PBMC feeder cells) are shown. Will be marked as M1015T and M1016T (1.3×10 5 cells) of two TIL lines with 46 different irradiated feeder cell batches (1.3 × 10 7 ), IL-2 (3000IU / mL), recombinant human IL-2 (eg, aldesleukin or equivalent) (CellGenix, Inc., Portsmouth, NH, USA) and OKT-3 (30ng / mL, MACS GMP CD3 pure, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) were co-cultured in T25 flasks for 7 days. Fold expansion values ​​of TILs were calculated on day 7. The graph shows the fold expansion values ​​for two TIL lines in separate stimulation experiments. For each TIL line, 4...

Embodiment 2

[0609] Example 2 - Selection of bone marrow cells for aAPC development

[0610] Phenotypic characterization of various myeloid lineage cell lines to identify potential candidates for further modification of aAPCs for TIL expansion. The results are summarized in Table 5. The MOLM-14 cell line exhibited endogenous expression of CD64 and was selected for further development. The EM-3 cell line was chosen based on the observed endogenous expression of ICOS-L (which was not observed in the EM-2 cell line although taken from the same patient).

[0611] Table 5. Summary of co-stimulatory molecules endogenously expressed on candidate cell lines of aAPC. CML refers to chronic myeloid leukemia and AML refers to acute myeloid leukemia. "Population" refers to the population of cells observed to express a marker (1 / 2 population = 50%).

[0612]

Embodiment 3

[0613] Embodiment 3-preparation of MOLM-14 artificial antigen-presenting cells (aMOLM14 aAPC)

[0614] MOLM-14 cells were obtained from Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH. To develop MOLM-14-based aAPCs, MOLM-14 cells were engineered with costimulatory molecules CD86 and 4-1BBL (CD137L). The human CD86 (hCD86) and human 4-1BBL (h4-1BBL) genes were cloned into commercially available PLV430G and transduced using the lentiviral transduction method with the PDONR221 vector (Invitrogen / ThermoFisher Scientific, Carlsbad, CA). , USA) co-transfection. The hCD86 and hCD137L genes were cloned into PLV430G and PDONR221 vectors using the Gateway cloning method (as described by Katzen, Expert Opin. Drug Disc. 2007, 4, 571-589). The 293T cell line (human embryonic kidney cells transformed with the large T antigen) was used for lentivirus production, transduced into MOLM-14 cells. Transfected cells were sorted (S3e Cell Sorter, Bio-Rad, H...

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Abstract

In some embodiments, compositions and methods relating to isolated artificial antigen presenting cells (aAPCs) are disclosed, including aAPCs comprising a myeloid cell transduced with one or more viral vectors, such as a MOLM-14 or a EM-3 myeloid cell, wherein the myeloid cell endogenously expresses HLA-A / B / C, ICOS-L, and CD58, and wherein the one or more viral vectors comprise a nucleic acid encoding CD86 and a nucleic acid encoding 4-lBBL and / or OX40L and transduce the myeloid cell to express CD86 and 4-lBBL and / or OX40L proteins. In some embodiments, methods of expanding tumor infiltratinglymphocytes (TILs) with aAPCs and methods of treating cancers using TILs after expansion with aAPCs are also disclosed.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Application No. 62 / 481,831 filed April 5, 2017, U.S. Provisional Application No. 62 / 475,053 filed March 22, 2017, and U.S. Provisional Application No. 62 / 475,053 filed December 23, 2016. 62 / 438,600 and priority to U.S. Provisional Application No. 62 / 415,274, filed October 31, 2016, the entire contents of which are incorporated herein by reference. technical field [0003] Engineered artificial antigen presenting cells (aAPCs) for the expansion of tumor infiltrating lymphocytes are disclosed. Background technique [0004] Treatment of bulky, refractory cancers using adoptive autologous transfer of tumor-infiltrating lymphocytes (TILs) represents an effective approach for treating patients with poor prognosis. See Gattinoni et al., Nat. Rev. Immunol. 2006, 6, 383-393. Successful immunotherapy requires large numbers of TILs, and commercialization requires robust and reliable meth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/078C12N5/0783A61K39/00
CPCC12N5/0634C12N5/0636C12N15/86C07K14/70532C07K14/70575A61K35/17A61P35/00C12N2502/11C12N2501/2302C12N2501/515C12N2510/00C12N2740/15043C12N5/0639C12N5/0635A61K39/4611A61K39/4644C12N5/065C12N2501/06C12N2501/51C12N2502/99C07K16/4283C12N2501/11C12N2501/25C12N2501/998A61K2039/515A61K2039/5158A61K39/0011C07K2317/622
Inventor A·维普塔朗A·戈克尔达斯B·拉比诺维奇M·T·洛策
Owner IOVANCE BIOTHERAPEUTICS INC
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