Unlock instant, AI-driven research and patent intelligence for your innovation.

Expansion of tils from cryopreserved tumor samples

A cryopreservation and tumor technology, applied in the field of tumor tissue expansion of lymphocytes, can solve the problems of high cost, limited potential for commercialization, high price, etc.

Active Publication Date: 2021-06-08
IOVANCE BIOTHERAPEUTICS INC
View PDF131 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Current TIL production methods are limited by length, cost, sterility issues, and other factors described herein, severely limiting the potential for such methods to be commercialized
One of the many limitations of current methods is the reliance on fresh tumor tissue to initiate production, which often requires costly and expensive local production facilities due to difficulties in transporting fresh tumors over longer distances

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Expansion of tils from cryopreserved tumor samples
  • Expansion of tils from cryopreserved tumor samples
  • Expansion of tils from cryopreserved tumor samples

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[1130] Adoptive cell transfer (ACT) is a highly effective form of immunotherapy that involves the transfer of immune cells with antitumor activity into cancer patients. ACT is a therapeutic approach that involves in vitro identification of lymphocytes with antitumor activity, massive expansion of these cells in vitro, and infusion into cancer-bearing hosts. Lymphocytes for adoptive transfer can be derived from the stroma of the resected tumor (tumor infiltrating lymphocytes or TIL). TILs for use in ACT can be prepared as described herein in Methods of TIL Preparation, including the use of tumor cryopreservation and thawing steps. In some embodiments, for example according to figure 1 method described in to prepare TIL. If they are genetically engineered to express anti-tumor T cell receptors (TCRs) or chimeric antigen receptors (CARs), enriched in mixed lymphocyte tumor cell cultures (MLTCs), or use autologous antigen-presenting cells and tumor-derived peptides clones, they...

Embodiment 1

[1193] Example 1: Preparation of media for pre-REP and REP processes

[1194] This example describes methods of preparing tissue culture media for use in protocols involving the culture of tumor-infiltrating lymphocytes (TILs) from various tumor types including, but not limited to, metastatic melanoma, head and neck Squamous cell carcinoma (HNSCC), ovarian cancer, triple negative breast cancer and lung adenocarcinoma. This medium can be used to prepare any of the TILs described in this application and in the Examples.

[1195] Preparation of CM1

[1196] The following reagents were removed from the freezer and heated in a 37°C water bath: RPMI1640, human AB serum, 200 mM L-glutamine. CM1 medium was prepared by adding each component to the top of a 0.2 μm filter unit in a volume suitable for filtration according to Table 21 below. Store at 4°C.

[1197] Table 21: Preparation of CM1

[1198] Element Final concentration Final volume 500ml Final volume 1L ...

Embodiment 2

[1209] Example 2: Use of IL-2, IL-15 and IL-21 cytokine cocktail

[1210] This example describes the IL-2, IL-15, and IL-21 cytokines ( acts as an additional T cell growth factor).

[1211] Using the methods described herein, TIL was grown from colorectal cancer tumors, melanoma tumors, cervical cancer tumors, triple negative breast tumors, lung tumors, and kidney tumors in the presence of IL-2 in one experimental arm; and In the other arm, IL-2 was replaced with IL-2, IL-15 and IL-21 at the start of the culture. After completion of pre-REP, cultures were assessed for expansion, phenotype, function (CD107a+ and IFN-γ) and TCRvβ pool. IL-15 and IL-21 are described elsewhere in this paper and in Gruijl et al. IL-21 promotes the expansion of CD27+CD28+ tumor-infiltrating lymphocytes with a high cytotoxic potential and a concomitant low expansion of regulatory T cells , Santegoets, S.J., J Transl Med., 2013, 11:37, available at https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC362...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

Methods of expanding tumor infiltrating lymphocytes (TILs) from cryopreserved tumor tissue and methods of using the expanded TILs in the treatment of human diseases, including cancers, are disclosed. Preparing the tumor tissue for freezing includes fragmenting the tumor tissue, and incubating the fragments in a cryopreservation medium. In an embodiment, the fragments may be incubated in the cryo preservation medium at 2 DEG C to 8 DEG C for about 30 to about 80 minutes. Freezing the fragments may be done by flash freezing using the vapor phase of liquid nitrogen, e.g. using a dry cryoshipper. In some embodiments, compositions of cryopreserved tumor tissues are disclosed.

Description

[0001] Related applications [0002] This International PCT Application claims priority to US Provisional Patent Application No. 62 / 733,937, filed on September 20, 2018, and US Provisional Patent Application No. 62 / 879,881, filed on July 29, 2019, the entire contents of each of which are hereby incorporated by reference. This article. technical field [0003] The invention described herein relates generally to the expansion of lymphocytes, and more specifically, but not exclusively, to the expansion of lymphocytes from cryopreserved tumor tissue. Background technique [0004] The use of adoptively transferred tumor infiltrating lymphocytes (TILs) to treat refractory cancers represents a potentially effective approach to the treatment of patients with poor prognosis. Gattinoni et al., Nat. Rev. Immunol., 2006, 6, 383-393. Large amounts of TILs are required for successful immunotherapy; therefore, robust and reliable methods are required for production and commercialization....

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02A61K35/17C12N5/078
CPCA01N1/0284C12N5/0636C12N2501/2302C12N2501/2315C12N2501/2321A61K39/4644A61K39/4611A61K35/17C12N5/0693A61P35/00C12N2501/515C12N2509/00A61K2039/505A01N1/0221C12N2509/10C12N2523/00
Inventor A·维普塔朗K·奥尼姆斯
Owner IOVANCE BIOTHERAPEUTICS INC