Methods, compositions, and implantable elements comprising active cells

An engineered active cell, wherein the engineered active cell is an engineered human retinal pigment epithelial (RPE) cell or an engineered cell derived from a human RPE cell, and wherein the engineered active cell comprises an exogenous nucleic acid encoding a polypeptide, wherein the exogenous nucleic acid comprises one or more of the following nucleotide sequences:

• (a) a promoter sequence which consists essentially of, or consists of (i) SEQ ID NO:23 or (ii) a sequence having at least 95%, 96%, 97%, 98%, 99% or greater sequence identity with SEQ ID NO:23;
• (b) a coding sequence encoding a Factor VIII-BDD (FVIII-BDD) polypeptide, wherein the FVIII-BDD polypeptide comprises, consists essentially of, or consists of SEQ ID NO:
• 1;
• SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:7 with an alanine instead of arginine at each of positions 787 and 790;
• (c) a coding sequence encoding a Factor IX (FIX) polypeptide, wherein the FIX polypeptide comprises, consists essentially of, or consists of SEQ ID NO:2;
• (d) a coding sequence encoding an interleukin-2 polypeptide (IL-2), wherein the IL-2 polypeptide comprises, consists essentially of, or consists of SEQ ID NO:29;
• (e) a coding sequence encoding a parathyroid hormone (PTH) polypeptide, wherein the PTH polypeptide comprises, consists essentially of, or consists of SEQ ID NO:30 or 31;
• (f) a coding sequence encoding a von Willebrand Factor (vWF) polypeptide, wherein the vWF polypeptide comprises, consists essentially of, or consists of SEQ ID NO:32 or 33;
• (g) a coding sequence encoding a conservatively substituted variant of an amino acid sequence in (b), (c), (d), (e) or (f);
• and
• (h) a coding sequence encoding an amino acid sequence that has as least 90%, 95%, 96%, 97%, 98%, 99% or greater sequence identity with the amino acid sequence in (b), (c), (d), (f) or (g).

An implantable element comprising an engineered active cell that produces or releases a therapeutic agent,

• wherein the engineered active cell has one or more of the following characteristics:
• (a) it comprises a human retinal pigment epithelial cell (RPE) or a cell derived therefrom;
• (b) it comprises a cell that has been obtained from a less differentiated cell;
• and
• (c) it comprises a cell that has one or more of the following properties:
• (i) it expresses one or more of the biomarkers CRALBP, RPE-65, RLBP, BEST1, or αB-crystallin;
• (ii) it does not express one or more of the biomarkers CRALBP, RPE-65, RLBP, BEST1, or αB-crystallin;
• (iii) it is naturally found in the retina and forms a monolayer above the choroidal blood vessels in the Bruch's membrane;
• and
• (iv) it is responsible for epithelial transport, light absorption, secretion, and/or immune modulation in the retina, and
• wherein the implantable element or an enclosing component thereof is modified with a compound of Formula (I):
• or a salt thereof, wherein:
• A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, —O—, —C(O)O—, —C(O)—, —OC(O)—, —N(RC)—, —N(RC)C(O)—, —C(O)N(RC)—, —N(RC)C(O)(C1-C6-alkylene)-, —N(RC)C(O)(C1-C6-alkenylene)-, —N(RC)N(RD)—, —NCN—, —C(═N(RC)(RD))O—, —S—, —S(O)x—, —OS(O)x, —N(RC)S(O)x, —S(O)xN(RC)—, —P(RF)y—, —Si(ORA)2—, —Si(RG)(ORA)—, —B(ORA)—, or a metal, wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is linked to an attachment group (e.g., an attachment group defined herein) and is optionally substituted by one or more R1;
• each of L1 and L3 is independently a bond, alkyl, or heteroalkyl, wherein each alkyl and heteroalkyl is optionally substituted by one or more R2;
• L2 is a bond;
• M is absent, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted by one or more R3;
• P is absent, cycloalkyl, heterocycyl, or heteroaryl each of which is optionally substituted by one or more R4;
• Z is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, —ORA, —C(O)RA, —C(O)ORA, —C(O)N(RC)(RD), —N(RC)C(O)RA, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted by one or more R5;
• each RA, RB, RC, RD, RE, RF, and RG is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with one or more R6;
• or RC and RD, taken together with the nitrogen atom to which they are attached, form a ring (e.g., a 5-7 membered ring), optionally substituted with one or more R6;
• each R1, R2, R3, R4, R5, and R6 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, —ORA1, —C(O)ORA1, —C(O)RB1, —OC(O)RB1, —N(RC1)(RD1), —N(RC1)C(O)RB1, —C(O)N(RC1), SRE1, S(O)xRE1, —OS(O)xRE1, —N(RC1)S(O)xRE1, —S(O)xN(RC1)(RD1), —P(RF1)y, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted by one or more R7;
• each RA1, RB1, RC1, RD1, RE1, and RF1 is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by one or more R7;
• each R7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxyl, cycloalkyl, or heterocyclyl;
• x is 1 or 2;
• and
• y is 2, 3, or 4.