Engineered Artificial Antigen Presenting Cells for Tumor Infiltrating Lymphocyte Expansion

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 4-1BBL and/or OX40L and transduce the myeloid cell to express CD86 and 4-1BBL and/or OX40L proteins. In some embodiments, methods of expanding tumor infiltrating lymphocytes (TILs) with aAPCs and methods of treating cancers using TILs after expansion with aAPCs are also disclosed.

1 .- 99 . (canceled) 
     
     
         100 . A method of treating a subject having cancer with a population of lymphocytes, the method comprising:
 (a) obtaining a first population of lymphocytes from a tumor resected from a patient;   (b) performing an initial expansion of the first population of lymphocytes in a first cell culture medium to obtain a second population of lymphocytes, wherein the second population of lymphocytes is at least 5-fold greater in number than the first population of lymphocytes, and wherein the first cell culture medium comprises IL-2;   (c) performing a rapid expansion of the second population of lymphocytes in a second cell culture medium to obtain a third population of lymphocytes, wherein the third population of lymphocytes is at least 50-fold greater in number than the second population of lymphocytes after about 7 days from the start of the rapid expansion; and wherein the second cell culture medium comprises IL-2 or OKT-3,   (d) transducing one of the first, the second, or the third population of lymphocytes with one or more viral vectors comprising a nucleic acid encoding a cell surface binding molecule, and/or one or more nucleic acids encoding one or more costimulatory molecules, wherein the first, the second, or the third population of lymphocytes expresses the cell surface binding molecule and the one or more costimulatory molecules; and   (e) administering a therapeutically effective portion of the third population of lymphocytes to a subject with the cancer.   
     
     
         101 . The method of  claim 100 , wherein the lymphocytes comprise tumor-infiltrating lymphocytes (TILs). 
     
     
         102 . The method of  claim 100 , wherein the one or more costimulatory molecules are independently selected from the group consisting of 4-1BB (CD137), OX40 (CD134), CD1a, CD1b, CD1c, CD1d, CD2, CD3γ, CD3δ, CD3∈, CD4, CD5, CD6, CD7, CD8α, CD8β, CD9, CD10, CD11a, CD11b, CD11c, CDw12, CD13, CD14, CD15, CD15s, CD16a, CD16b, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD33, CD34, CD35, CD3δ, CD37, CD38, CD39, CD40, CD41, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD45R, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CDw60, CD61, CD62E, CD62L, CD62P, CD63, CD64, CD65, CD66a, CD66b, CD66c, CD66d, CD66e, CD66f, CD67, CD68, CD69, CDw70, CD71, CD72, CD73, CD74, CDw75, CDw76, CD77, CD79α, CD79β, CD80, CD81, CD82, CD83, CD84, CD85, CD86, CD87, CD88, CD89, CD90, CD91, CDw92, CD93, CD94, CD95, CD96, CD97, CD98, CD99, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107a, CD107b, CDw108, CDw109, CD114, CD115, CD116, CD117, CD118, CD119, CD120a, CD120b, CD121a, CD121b, CD122, CD123, CDw124, CD125, CD126, CDw127, CDw128a, CDw128b, CDw130, CDw131, CD132, CD133, CD135, CD136, CD138, CD139, CD140a, CD140b, CD141, CD142, CD143, CD144, CDw145, CD146, CD147, CD148, CDw149, CD150, CD151, CD152, CD153, CD154, CD155, CD156, CD157, CD158a, CD158b, CD161, CD162, CD163, CD164, CD165, CD166, and TCRξ. 
     
     
         103 . The method of  claim 102 , wherein the one or more costimulatory molecules are independently selected from the group consisting of CD28, 4-1BB (CD137), and OX40 (CD134). 
     
     
         104 . The method of  claim 100 , wherein the cell culture medium comprises IL-2. 
     
     
         105 . The method of  claim 104 , wherein the IL-2 is at an initial concentration of about 3000 IU/mL. 
     
     
         106 . The method of  claim 100 , wherein the cell culture medium comprises OKT-3 antibody. 
     
     
         107 . The method of  claim 106 , wherein the OKT-3 antibody is at an initial concentration of about 30 ng/mL. 
     
     
         108 . The method of  claim 100 , wherein the rapid expansion is performed over a period not greater than 14 days. 
     
     
         109 . The method of  claim 100 , wherein one or both of the initial expansion and the rapid expansion is performed using a gas permeable container. 
     
     
         110 . The method of  claim 100 , wherein the cancer is selected from the group consisting of melanoma, ovarian cancer, cervical cancer, non-small-cell lung cancer (NSCLC), lung cancer, bladder cancer, breast cancer, cancer caused by human papilloma virus, head and neck cancer, renal cancer, renal cell carcinoma, pancreatic cancer, and glioblastoma. 
     
     
         111 . The method of  claim 100 , wherein the one or more viral vectors comprise a lentiviral vector. 
     
     
         112 . The method of  claim 100 , wherein the rapid expansion is performed using a population of antigen presenting cells (APCs). 
     
     
         113 . The method of  claim 112 , wherein the population of APCs expands the population of lymphocytes by at least 50-fold over a period of about 7 days. 
     
     
         114 . The method of  claim 112 , wherein the population of APCs endogenously express HLA-AB/C, ICOS-L, and CD58. 
     
     
         115 . The method of  claim 112 , wherein the population of APCs are transduced to express an anti-OKT-3 antibody scFv binding domain. 
     
     
         116 . The method of  claim 112 , wherein the ratio of the second population of lymphocytes to the population of APCs is between about 1 to 200 and about 1 to 400. 
     
     
         117 . The method of  claim 100 , wherein the population of lymphocytes are cryopreserved. 
     
     
         118 . The method of  claim 101 , wherein the TILs are cryopreserved. 
     
     
         119 . The method of  claim 100 , wherein the cell surface binding molecule comprises a single chain fragment variable (scFv) binding domain.