T cell receptors targeting an NPM1 neoantigen

Abstract

Provided herein are T cell receptors (TCRs) or antigen-binding fragments thereof, such as those that recognize or bind a NPMlc neoantigen. In particular, the present disclosure relates to TCRs that bind or recognize particular NPMlc peptides in the context of a major histocompatibility complex (MHC) molecule. The present disclosure further relates to nucleic acids encoding such TCRs, engineered cells comprising such TCRs, methods of isolating such TCRs, and uses thereof, for example, in cell therapy.

Description

T Cell Receptors Targeting an NPM1 NeoantigenCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 580,503, filed September 5, 2023, and U.S. Provisional Application No. 63 / 651,653, filed May 24, 2024, each of which is incorporated herein by reference.SEQUENCE LISTING

[0002] The Sequence Listing written in file BSB-0009W001_SeqList.xml is 383 kilobytes in size, was created September 3, 2024, and is hereby incorporated by reference.BACKGROUND

[0003] Allogeneic hematopoietic stem cell transplantation (HSCT) (HLA-matched or HLA haplotype matched) may be used for treatment of diseases or conditions such as hematologic malignancies and other nonmalignant conditions. Some subjects may relapse after alloSCT or are ineligible for alloSCT. Improved treatments are necessary to attain an optimal treatment outcome. Provided are T cell receptors (TCRs) and engineered T cells containing the TCRs that meet such needs.SUMMARY

[0004] Described are anti-NPMlc T cell receptors (anti-AIQDLCLAV TCRs and anti- CLAVEEVSL TCRs), or antigen-binding fragments thereof, that recognize or bind an NPMlc neoantigen (i.e., AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5)). In some embodiments, the described anti-NPMlc TCRs bind to or recognize the peptide AIQDLCLAV (SEQ ID NO: 1). In some embodiments, the described anti-NPMlc TCRs bind to or recognize the peptide CLAVEEVSL (SEQ ID NO:5). In some embodiments, the described anti-NPMlc TCRs bind to or recognize the peptide AIQDLCLAV (SEQ ID NO: 1) in the context of a major histocompatibility complex (MHC) molecule. In some embodiments, the described anti- NPMlc TCRs bind to or recognize the peptide CLAVEEVSL (SEQ ID NO: 5) in the context of a major histocompatibility complex (MHC) molecule. The present disclosure further relates to polynucleotides encoding such anti-NPMlc TCRs, purified anti-NPMlc TCRs, purified anti-NPMlc TCR-anti-CD3 fusion proteins, engineered cells comprising or expression such anti-NPMlc TCRs, methods of isolating such anti-NPMlc TCRs and uses thereof, for example, in T cell therapy, e.g., engineered autologous T cell therapy.

[0005] Provided herein are anti-NPMlc TCRs or antigen-binding fragment thereof,comprising: an a chain comprising a variable a (Va) region and a P chain comprising a variable P (VP) region; or a y chain comprising a variable y (Vy) region and a 5 chain comprising a variable 5 (V5) region; wherein:(a) the Va or Vy region comprises a complementarity determining region 3 (CDR-3) comprising SEQ ID NO: 13, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:21;(b) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:31, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:39;(c) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:49, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:57;(d) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:67, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:75;(e) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:85, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:93;(f) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 103, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 111;(g) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 121, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 129;(h) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 139, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 147;(i) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 157, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 165;(j) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 175, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 183;(k) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 193, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:201;(l) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:211, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:219;(m) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:229, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:237;(n) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:247, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:255;(o) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:265, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:273; or(р) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:283, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:291.Anti-NPMlc TRCs corresponding any of (a)-(c) as described above recognize the AIQDLCLAV (SEQ ID NO: 1) peptide. Anti-NPMlc TRCs corresponding any of (d)-(p) as described above recognize the CLAVEEVSL (SEQ ID NO:5) peptide.

[0006] Also provided herein are anti-NPMlc TCRs or antigen-binding fragment thereof, comprising: an a chain comprising a Va region and a P chain comprising a VP region; or a y chain comprising a Vy region and a 5 chain comprising a V5 region; wherein:(a) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 11, a CDR-2 comprising SEQ ID NO: 12, and a CDR-3 comprising SEQ ID NO: 13, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 19, a CDR-2 comprising SEQ ID NO:20, and a CDR-3 comprising SEQ ID NO:21;(b) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:29, a CDR-2 comprising SEQ ID NO:30, and a CDR-3 comprising SEQ ID NO:31, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:37, a CDR-2 comprising SEQ ID NO:38, and a CDR-3 comprising SEQ ID NO:39;(с) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:47, a CDR-2 comprising SEQ ID NO:48, and a CDR-3 comprising SEQ ID NO:49, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:55, a CDR-2 comprising SEQ ID NO:56, and a CDR-3 comprising SEQ ID NO:57;(d) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:65, a CDR-2 comprising SEQ ID NO:66, and a CDR-3 comprising SEQ ID NO:67, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:73, a CDR-2 comprising SEQ ID NO:74, and a CDR-3 comprising SEQ ID NO:75;(e) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:83, a CDR-2 comprising SEQ ID NO:84, and a CDR-3 comprising SEQ ID NO:85, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:91, a CDR-2 comprising SEQ ID NO:92, and a CDR-3 comprising SEQ ID NO:93;(f) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR-2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111;(g) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and the VP or V5region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129;(h) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 137, a CDR-2 comprising SEQ ID NO: 138, and a CDR-3 comprising SEQ ID NO: 139, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 145, a CDR-2 comprising SEQ ID NO: 146, and a CDR-3 comprising SEQ ID NO: 147;(i) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 155, a CDR-2 comprising SEQ ID NO: 156, and a CDR-3 comprising SEQ ID NO: 157, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 163, a CDR-2 comprising SEQ ID NO: 164, and a CDR-3 comprising SEQ ID NO: 165;(j) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 173, a CDR-2 comprising SEQ ID NO: 174, and a CDR-3 comprising SEQ ID NO: 175, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 181, a CDR-2 comprising SEQ ID NO: 182, and a CDR-3 comprising SEQ ID NO: 183;(k) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ IDNO:200, and a CDR-3 comprising SEQ ID NO:201;(l) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:209, a CDR-2 comprising SEQ ID NO:210, and a CDR-3 comprising SEQ ID NO:211, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO :217, a CDR-2 comprising SEQ IDNO:218, and a CDR-3 comprising SEQ ID NO:219;(m) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:227, a CDR-2 comprising SEQ ID NO:228, and a CDR-3 comprising SEQ ID NO:229, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:235, a CDR-2 comprising SEQ IDNO:236, and a CDR-3 comprising SEQ ID NO:237;(n) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:245, a CDR-2 comprising SEQ ID NO:246, and a CDR-3 comprising SEQ ID NO:247, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:253, a CDR-2 comprising SEQ IDNO:254, and a CDR-3 comprising SEQ ID NO:255;(o) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:263, a CDR-2 comprising SEQ ID NO:264, and a CDR-3 comprising SEQ ID NO:265, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:271, a CDR-2 comprising SEQ IDNO:272, and a CDR-3 comprising SEQ ID NO:273; or(р) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:281, a CDR-2 comprising SEQ ID NO:282, and a CDR-3 comprising SEQ ID NO:283, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:289, a CDR-2 comprising SEQ IDNO:290, and a CDR-3 comprising SEQ ID NO:291.

[0007] Also provided herein are anti-NPMlc TCRs or antigen-binding fragment thereof, comprising: an a chain comprising a Va region and a P chain comprising a VP region; or a y chain comprising a Vy region and a 5 chain comprising a V5 region; wherein:(a) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:22; (b) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:40;(с) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:50, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:58;(d) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:68, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:76;(e) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:86, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:94;(f) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 104, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO: 112;(g) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 122, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO: 130;(h) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 140, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO: 148;(i) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 158, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO: 166;(j) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 176, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO: 184;(k) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 194, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:202;(l) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:212, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:220;(m) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:230, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:238;(n) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:248, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:256;(o) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:266, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:274; or(p) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:284, and the VP or V5 region comprises a CDR-3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:292.

[0008] In some embodiments,(a) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:22;(b) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:40;(a) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:22;(b) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:40;(c) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:50, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:58;(d) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:68, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:76;(e) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:86, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:94;(f) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 104, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 112;(g) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 122, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 130;(h) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 140, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 148;(i) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 158, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 166;(j) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 176, andthe VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 184;(k) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 194, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:202;(l) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:212, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:220;(m) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:230, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:238;(n) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:248, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:256;(o) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:266, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:274; or(p) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:284, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:292.

[0009] Also provided are anti-NPMlc TCRs or antigen-binding fragment thereof, comprising: an a chain comprising a Va region and a P chain comprising a VP region; or a y chain comprising a Vy region and a 5 chain comprising a V5 region; wherein:(a) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:22;(b) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:40;(c) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:50, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:58;(d) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:68, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 76;(e) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:86, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 94;(f) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 104, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 112;(g) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 122, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 130;(h) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 140, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequenceof SEQ ID NO: 148;(i) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 158, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 166;(j) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 176, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO: 184;(k) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 194, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:202;(l) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:212, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:220;(m) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:230, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:238;(n) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:248, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:256;(o) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:266, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprisinga CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:274; or(р) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR- 1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:284, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:292.

[0010] Also provided are anti-NPMlc TCRs or antigen-binding fragments thereof, comprising: an a chain comprising a Va region and a P chain comprising a VP region, or a y chain comprising a Vy region and a 5 chain comprising a V5 region, wherein:(a) the Va or Vy region comprises SEQ ID NO: 14 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:22 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 11, a CDR-2 comprising SEQ ID NO: 12, and a CDR-3 comprising SEQ ID NO: 13, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 19, a CDR-2 comprising SEQ ID NO:20, and a CDR-3 comprising SEQ ID NO:21;(b) the Va or Vy region comprises SEQ ID NO:32 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:40 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:29, a CDR-2 comprising SEQ ID NO:30, and a CDR-3 comprising SEQ ID NO:31, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:37, a CDR-2 comprising SEQ ID NO:38, and a CDR-3 comprising SEQ ID NO:38;(с) the Va or Vy region comprises SEQ ID NO:50 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:58 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:47, a CDR-2 comprising SEQ ID NO:48, and a CDR-3 comprising SEQ ID NO:49, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:55, a CDR-2 comprising SEQ ID NO:56, and a CDR-3 comprising SEQ ID NO:57;(d) the Va or Vy region comprises SEQ ID NO:68 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:76 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:65, a CDR-2 comprising SEQ ID NO:66, and a CDR-3 comprising SEQ ID NO:67, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:73, a CDR-2 comprising SEQ ID NO:74, and a CDR-3 comprising SEQ ID NO:75;(e) the Va or Vy region comprises SEQ ID NO:86 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:94 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:83, a CDR-2 comprising SEQ ID NO:84, and a CDR-3 comprising SEQ ID NO:85, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:91, a CDR-2 comprising SEQ ID NO:92, and a CDR-3 comprising SEQ ID NO:93;(f) the Va or Vy region comprises SEQ ID NO: 104 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 112 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR-2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111;(g) the Va or Vy region comprises SEQ ID NO: 122 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 130 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129;(h) the Va or Vy region comprises SEQ ID NO: 140 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 148 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 137, a CDR-2 comprising SEQ ID NO: 138, and a CDR-3 comprising SEQ ID NO: 139, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 145, a CDR-2 comprising SEQ ID NO: 146, and a CDR-3 comprising SEQ ID NO: 147;(i) the Va or Vy region comprises SEQ ID NO: 158 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 166 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 155, a CDR-2 comprising SEQ ID NO: 156, and a CDR-3 comprising SEQ ID NO: 157, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 163, a CDR-2 comprising SEQ ID NO: 164, and a CDR-3 comprising SEQ ID NO: 165;(j) the Va or Vy region comprises SEQ ID NO: 176 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 184 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 173, a CDR-2 comprising SEQ ID NO: 174, and a CDR-3 comprisingSEQ ID NO: 175, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 181, a CDR-2 comprising SEQ ID NO: 182, and a CDR-3 comprising SEQ ID NO: 183;(k) the Va or Vy region comprises SEQ ID NO: 194 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:202 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ ID NO:200, and a CDR-3 comprising SEQ ID NO:201;(l) the Va or Vy region comprises SEQ ID NO:212 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:220 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:209, a CDR-2 comprising SEQ IDNO:210, and a CDR-3 comprising SEQ ID NO:211, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:217, a CDR-2 comprising SEQ ID NO:218, and a CDR-3 comprising SEQ ID NO:219;(m) the Va or Vy region comprises SEQ ID NO:230 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:238 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:227, a CDR-2 comprising SEQ ID NO:228, and a CDR-3 comprising SEQ ID NO:229, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:235, a CDR-2 comprising SEQ ID NO:236, and a CDR-3 comprising SEQ ID NO:23;(n) the Va or Vy region comprises SEQ ID NO:248 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:256 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:245, a CDR-2 comprising SEQ ID NO:246, and a CDR-3 comprising SEQ ID NO:247, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:253, a CDR-2 comprising SEQ ID NO:254, and a CDR-3 comprising SEQ ID NO:255;(o) the Va or Vy region comprises SEQ ID NO:266 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:274 or a sequence that has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:263, a CDR-2 comprising SEQ ID NO:264, and a CDR-3 comprising SEQ ID NO:265, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:271, a CDR-2 comprising SEQ ID NO:272, and a CDR-3 comprising SEQ ID NO:273; or(p) the Va or Vy region comprises SEQ ID NO:284 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:292 or a sequencethat has at least 90% sequence identity thereto, wherein the Va or Vy region comprises a CDR- 1 comprising SEQ ID NO:281, a CDR-2 comprising SEQ IDNO:282, and a CDR-3 comprising SEQ ID NO:283, and wherein the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:289, a CDR-2 comprising SEQ ID NO:290, and a CDR-3 comprising SEQ ID NO:291.

[0011] In some embodiments,(a) the Va or Vy region comprises SEQ ID NO: 14, and the VP or V5 region comprises SEQ ID NO:22;(b) the Va or Vy region comprises SEQ ID NO:32, and the VP or V5 region comprisesSEQ ID NO:40;(c) the Va or Vy region comprises SEQ ID NO:50, and the VP or V5 region comprises SEQ ID NO:58;(d) the Va or Vy region comprises SEQ ID NO:68, and the VP or V5 region comprisesSEQ ID NO: 76;(e) the Va or Vy region comprises SEQ ID NO:86, and the VP or V5 region comprises SEQ ID NO: 94;(f) the Va or Vy region comprises SEQ ID NO: 104, and the VP or V5 region comprises SEQ ID NO: 112;(g) the Va or Vy region comprises SEQ ID NO: 122, and the VP or V5 region comprises SEQ ID NO: 130;(h) the Va or Vy region comprises SEQ ID NO: 140, and the VP or V5 region comprises SEQ ID NO: 148;(i) the Va or Vy region comprises SEQ ID NO: 158, and the VP or V5 region comprises SEQ ID NO: 166;(j) the Va or Vy region comprises SEQ ID NO: 176, and the VP or V5 region comprises SEQ ID NO: 184;(k) the Va or Vy region comprises SEQ ID NO: 194, and the VP or V5 region comprises SEQ ID NO:202;(l) the Va or Vy region comprises SEQ ID NO:212, and the VP or V5 region comprises SEQ ID NO:220;(m) the Va or Vy region comprises SEQ ID NO:230, and the VP or V5 region comprises SEQ ID NO:238;(n) the Va or Vy region comprises SEQ ID NO:248, and the VP or V5 region comprises SEQ ID NO:256;(o) the Va or Vy region comprises SEQ ID NO:266, and the VP or V5 regioncomprises SEQ ID NO:274; or(р) the Va or Vy region comprises SEQ ID NO:284, and the VP or V5 region comprises SEQ ID NO:292.

[0012] In some embodiments, the a chain further comprises an a constant (Ca) region and the P chain further comprises a P constant (CP) region; or the y chain further comprises an y constant (Cy) region and the 5 chain further comprises a 5 constant (C5) region. In some embodiments, the Ca comprises SEQ ID NO:3, and the CP comprises SEQ ID NO:7.

[0013] In some embodiments,(a) the a or y chain comprises SEQ ID NO: 17, and the P or 5 chain comprises SEQ ID NO:25;(b) the a or y chain comprises SEQ ID NO:35, and the P or 5 chain comprises SEQ ID NO:43;(с) the a or y chain comprises SEQ ID NO:53, and the P or 5 chain comprises SEQ ID NO:61;(d) the a or y chain comprises SEQ ID NO:71, and the P or 5 chain comprises SEQ ID NO: 79;(e) the a or y chain comprises SEQ ID NO:89, and the P or 5 chain comprises SEQ ID NO: 97;(f) the a or y chain comprises SEQ ID NO: 107, and the P or 5 chain comprises SEQ ID NO: 115;(g) the a or y chain comprises SEQ ID NO: 125, and the P or 5 chain comprises SEQ ID NO: 133;(h) the a or y chain comprises SEQ ID NO: 143, and the P or 5 chain comprises SEQ ID NO: 151;(i) the a or y chain comprises SEQ ID NO: 161, and the P or 5 chain comprises SEQ ID NO: 169;(j) the a or y chain comprises SEQ ID NO: 179, and the P or 5 chain comprises SEQ ID NO: 187;(k) the a or y chain comprises SEQ ID NO: 197, and the P or 5 chain comprises SEQ ID NO:205;(l) the a or y chain comprises SEQ ID NO:215, and the P or 5 chain comprises SEQ ID NO:223;(m) the a or y chain comprises SEQ ID NO:233, and the P or 5 chain comprises SEQ ID NO:241;(n) the a or y chain comprises SEQ ID NO:251, and the P or 5 chain comprises SEQ ID NO:259;(o) the a or y chain comprises SEQ ID NO:269, and the P or 5 chain comprises SEQ ID NO:277; or(p) the a or y chain comprises SEQ ID NO:287, and the P or 5 chain comprises SEQ ID NO:295.

[0014] In some embodiments, the anti-NPMlc TCR or antigen-binding fragment thereof recognizes a NPMlc neoantigen in the context of an MHC molecule. In some embodiments, the MHC molecule is a human leukocyte antigen (HLA)-A molecule. In some embodiments, the HLA-A molecule is of serotype HLA-A*02:01. In some embodiments, the HLA-A molecule is of serotype HLA-A*02:06. In some embodiments, the HLA-A molecule is of serotype HLA-A*02:03. In some embodiments, the neoantigen of NPMlc is set forth in SEQ ID NO: 1 or 5.

[0015] Also provided are nucleotide sequences (e.g., polynucleotides or nucleic acids) encoding any of the anti-NPMlc TCRs or antigen-binding fragment thereof described herein, or a Va, a Vy, a VP, a V5, an a chain, a P chain, a y chain, or a 5 chain thereof. The nucleotide sequences can be provided as a pair of nucleotide sequences comprising a first nucleotide sequence encoding a Va region or an a chain and a second nucleotide sequence encoding a VP region or a P chain; or a first nucleotide sequence encoding a Vy region or a y chain and a second nucleotide sequence encoding a V5 region or a 5 chain. The first and second nucleotide sequences can be provided on separate expression cassettes, plasmids, or vectors e.g., viral vectors) or on a single expression cassette, plasmid, or vector. If provided on a single expression cassette, plasmid, or vector, the first and second nucleotide sequences can be linked by a 2A element (e.g., a P2A element) or an internal ribosome entry sequence (IRES) and expressed from a single promoter.

[0016] In some embodiments, nucleotide sequences encoding an anti-NPMlc TCR, or an antigen binding fragment thereof, comprise a first nucleotide sequence encoding the Va region and a second nucleotide sequence encoding the VP region; or a first nucleotide sequence encoding the Vy region and a second nucleotide sequence encoding the V5 region; wherein:(a) the first nucleotide sequence encodes SEQ ID NO: 14 and the second nucleotide sequence encodes SEQ ID NO:22;(b) the first nucleotide sequence encodes SEQ ID NO:32 and the second nucleotide sequence encodes SEQ ID NO:40;(c) the first nucleotide sequence encodes SEQ ID NO:50 and the second nucleotidesequence encodes SEQ ID NO:58;(d) the first nucleotide sequence encodes SEQ ID NO: 68 and the second nucleotide sequence encodes SEQ ID NO:76;(e) the first nucleotide sequence encodes SEQ ID NO: 86 and the second nucleotide sequence encodes SEQ ID NO: 94;(f) the first nucleotide sequence encodes SEQ ID NO: 104 and the second nucleotide sequence encodes SEQ ID NO: 112;(g) the first nucleotide sequence encodes SEQ ID NO: 122 and the second nucleotide sequence encodes SEQ ID NO: 130;(h) the first nucleotide sequence encodes SEQ ID NO: 140 and the second nucleotide sequence encodes SEQ ID NO: 148;(i) the first nucleotide sequence encodes SEQ ID NO: 158 and the second nucleotide sequence encodes SEQ ID NO: 166;(j) the first nucleotide sequence encodes SEQ ID NO: 176 and the second nucleotide sequence encodes SEQ ID NO: 184;(k) the first nucleotide sequence encodes SEQ ID NO: 194 and the second nucleotide sequence encodes SEQ ID NO:202;(l) the first nucleotide sequence encodes SEQ ID NO:212 and the second nucleotide sequence encodes SEQ ID NO:220;(m) the first nucleotide sequence encodes SEQ ID NO:230 and the second nucleotide sequence encodes SEQ ID NO:238;(n) the first nucleotide sequence encodes SEQ ID NO:248 and the second nucleotide sequence encodes SEQ ID NO:256;(o) the first nucleotide sequence encodes SEQ ID NO:266 and the second nucleotide sequence encodes SEQ ID NO:274; or(p) the first nucleotide sequence encodes SEQ ID NO:284 and the second nucleotide sequence encodes SEQ ID NO:292.Anti-NPMlc TRCs corresponding any of (a)-(c) as described above recognize the AIQDLCLAV (SEQ ID NO: 1) peptide. Anti-NPMlc TRCs corresponding any of (d)-(p) as described above recognize the CLAVEEVSL (SEQ ID NO: 5) peptide.

[0017] In some embodiments,(a) the first nucleotide sequence comprises SEQ ID NO: 15 or a sequence that encodes SEQ ID NO: 14 and has at least 90% sequence identity to SEQ ID NO: 15, and the second nucleotide sequence comprises SEQ ID NO:23 or a sequence that encodes SEQ ID NO:22 andhas at least 90% sequence identity to SEQ ID NO:23;(b) the first nucleotide sequence comprises SEQ ID NO:33 or a sequence that encodes SEQ ID NO:32 and has at least 90% sequence identity to SEQ ID NO:33, and the second nucleotide sequence comprises SEQ ID NO:41 or a sequence that encodes SEQ ID NO:40 and has at least 90% sequence identity to SEQ ID NO:41.(c) the first nucleotide sequence comprises SEQ ID NO:51 or a sequence that encodes SEQ ID NO:50 and has at least 90% sequence identity to SEQ ID NO:51, and the second nucleotide sequence comprises SEQ ID NO:59 or a sequence that encodes SEQ ID NO:58 and has at least 90% sequence identity to SEQ ID NO:59;(d) the first nucleotide sequence comprises SEQ ID NO:69 or a sequence that encodes SEQ ID NO:68 and has at least 90% sequence identity to SEQ ID NO:69, and the second nucleotide sequence comprises SEQ ID NO:77 or a sequence that encodes SEQ ID NO:76 and has at least 90% sequence identity to SEQ ID NO:77;(e) the first nucleotide sequence comprises SEQ ID NO:87 or a sequence that encodes SEQ ID NO:86 and has at least 90% sequence identity to SEQ ID NO:87, and the second nucleotide sequence comprises SEQ ID NO:95 or a sequence that encodes SEQ ID NO:94 and has at least 90% sequence identity to SEQ ID NO: 95;(f) the first nucleotide sequence comprises SEQ ID NO: 105 or a sequence that encodes SEQ ID NO: 104 and has at least 90% sequence identity to SEQ ID NO: 105, and the second nucleotide sequence comprises SEQ ID NO: 113 or a sequence that encodes SEQ ID NO: 112 and has at least 90% sequence identity to SEQ ID NO: 113;(g) the first nucleotide sequence comprises SEQ ID NO: 123 or a sequence that encodes SEQ ID NO: 122 and has at least 90% sequence identity to SEQ ID NO: 123, and the second nucleotide sequence comprises SEQ ID NO: 131 or a sequence that encodes SEQ ID NO: 1130 and has at least 90% sequence identity to SEQ ID NO: 131;(h) the first nucleotide sequence comprises SEQ ID NO: 141 or a sequence that encodes SEQ ID NO: 140 and has at least 90% sequence identity to SEQ ID NO: 141, and the second nucleotide sequence comprises SEQ ID NO: 149 or a sequence that encodes SEQ ID NO: 148 and has at least 90% sequence identity to SEQ ID NO: 149;(i) the first nucleotide sequence comprises SEQ ID NO: 159 or a sequence that encodes SEQ ID NO: 158 and has at least 90% sequence identity to SEQ ID NO: 159, and the second nucleotide sequence comprises SEQ ID NO: 167 or a sequence that encodes SEQ ID NO: 166 and has at least 90% sequence identity to SEQ ID NO: 167;(j) the first nucleotide sequence comprises SEQ ID NO: 177 or a sequence thatencodes SEQ ID NO: 176 and has at least 90% sequence identity to SEQ ID NO: 177, and the second nucleotide sequence comprises SEQ ID NO: 185 or a sequence that encodes SEQ ID NO: 184 and has at least 90% sequence identity to SEQ ID NO: 185;(k) the first nucleotide sequence comprises SEQ ID NO: 195 or a sequence that encodes SEQ ID NO: 194 and has at least 90% sequence identity to SEQ ID NO: 195, and the second nucleotide sequence comprises SEQ ID NO:203 or a sequence that encodes SEQ ID NO:202 and has at least 90% sequence identity to SEQ ID NO:203;(l) the first nucleotide sequence comprises SEQ ID NO:213 or a sequence that encodes SEQ ID NO:212 and has at least 90% sequence identity to SEQ ID NO:213, and the second nucleotide sequence comprises SEQ ID NO:221 or a sequence that encodes SEQ ID NO:220 and has at least 90% sequence identity to SEQ ID NO:221;(m) the first nucleotide sequence comprises SEQ ID NO:231 or a sequence that encodes SEQ ID NO:230 and has at least 90% sequence identity to SEQ ID NO:231, and the second nucleotide sequence comprises SEQ ID NO:239 or a sequence that encodes SEQ ID NO:238 and has at least 90% sequence identity to SEQ ID NO:239;(n) the first nucleotide sequence comprises SEQ ID NO:249 or a sequence that encodes SEQ ID NO:248 and has at least 90% sequence identity to SEQ ID NO:249, and the second nucleotide sequence comprises SEQ ID NO:257 or a sequence that encodes SEQ ID NO:256 and has at least 90% sequence identity to SEQ ID NO:257;(o) the first nucleotide sequence comprises SEQ ID NO:267 or a sequence that encodes SEQ ID NO:266 and has at least 90% sequence identity to SEQ ID NO:267, and the second nucleotide sequence comprises SEQ ID NO:275 or a sequence that encodes SEQ ID NO:274 and has at least 90% sequence identity to SEQ ID NO:275; or(p) the first nucleotide sequence comprises SEQ ID NO:285 or a sequence that encodes SEQ ID NO:284 and has at least 90% sequence identity to SEQ ID NO:285, and the second nucleotide sequence comprises SEQ ID NO:293 or a sequence that encodes SEQ ID NO:292 and has at least 90% sequence identity to SEQ ID NO:293.

[0018] In some embodiments, the one of more of the nucleic acid sequences encoding the Va, Vy, VP, V5, a constant, y constant, P constant, or 5 constant chains is codon optimized. In some embodiments,(a) the first nucleotide sequence comprises SEQ ID NO: 16, and the second nucleotide sequence comprises SEQ ID NO:24;(b) the first nucleotide sequence comprises SEQ ID NO:34, and the second nucleotide sequence comprises SEQ ID NO:42;(c) the first nucleotide sequence comprises SEQ ID NO:52, and the second nucleotide sequence comprises SEQ ID NO:60(d) the first nucleotide sequence comprises SEQ ID NO:70, and the second nucleotide sequence comprises SEQ ID NO:78;(e) the first nucleotide sequence comprises SEQ ID NO:88, and the second nucleotide sequence comprises SEQ ID NO:96;(f) the first nucleotide sequence comprises SEQ ID NO: 106, and the second nucleotide sequence comprises SEQ ID NO: 114;(g) the first nucleotide sequence comprises SEQ ID NO: 124, and the second nucleotide sequence comprises SEQ ID NO: 132;(h) the first nucleotide sequence comprises SEQ ID NO: 142, and the second nucleotide sequence comprises SEQ ID NO: 150;(i) the first nucleotide sequence comprises SEQ ID NO: 160, and the second nucleotide sequence comprises SEQ ID NO: 168;(j) the first nucleotide sequence comprises SEQ ID NO: 178, and the second nucleotide sequence comprises SEQ ID NO: 186;(k) the first nucleotide sequence comprises SEQ ID NO: 196, and the second nucleotide sequence comprises SEQ ID NO:204;(l) the first nucleotide sequence comprises SEQ ID NO:214, and the second nucleotide sequence comprises SEQ ID NO:222;(m) the first nucleotide sequence comprises SEQ ID NO:232, and the second nucleotide sequence comprises SEQ ID NO:240;(n) the first nucleotide sequence comprises SEQ ID NO:250, and the second nucleotide sequence comprises SEQ ID NO:258;(o) the first nucleotide sequence comprises SEQ ID NO:268, and the second nucleotide sequence comprises SEQ ID NO:276; or(p) the first nucleotide sequence comprises SEQ ID NO:286, and the second nucleotide sequence comprises SEQ ID NO:294.

[0019] In some embodiments, nucleotide sequences encoding an anti-NPMlc TCR, or an antigen binding fragment thereof, comprise a first nucleotide sequence encoding the a chain and a second nucleotide sequence encoding a P chain; or a first nucleotide sequence encoding an y chain and a second nucleotide sequence encoding a 5 chain; wherein:(a) the first nucleotide sequence encodes SEQ ID NO: 17, and the second nucleotide sequence encodes SEQ ID NO:25;(b) the first nucleotide sequence encodes SEQ ID NO:35, and the second nucleotide sequence encodes SEQ ID NO:43;(c) the first nucleotide sequence encodes SEQ ID NO:53, and the second nucleotide sequence encodes SEQ ID NO:61;(d) the first nucleotide sequence encodes SEQ ID NO:71, and the second nucleotide sequence encodes SEQ ID NO:79;(e) the first nucleotide sequence encodes SEQ ID NO:89, and the second nucleotide sequence encodes SEQ ID NO: 97;(f) the first nucleotide sequence encodes SEQ ID NO: 107, and the second nucleotide sequence encodes SEQ ID NO: 115;(g) the first nucleotide sequence encodes SEQ ID NO: 125, and the second nucleotide sequence encodes SEQ ID NO: 133;(h) the first nucleotide sequence encodes SEQ ID NO: 143, and the second nucleotide sequence encodes SEQ ID NO: 151;(i) the first nucleotide sequence encodes SEQ ID NO: 161, and the second nucleotide sequence encodes SEQ ID NO: 169;(j) the first nucleotide sequence encodes SEQ ID NO: 179, and the second nucleotide sequence encodes SEQ ID NO: 187;(k) the first nucleotide sequence encodes SEQ ID NO: 197, and the second nucleotide sequence encodes SEQ ID NO:205;(l) the first nucleotide sequence encodes SEQ ID NO:215, and the second nucleotide sequence encodes SEQ ID NO:223;(m) the first nucleotide sequence encodes SEQ ID NO:233, and the second nucleotide sequence encodes SEQ ID NO:241;(n) the first nucleotide sequence encodes SEQ ID NO:251, and the second nucleotide sequence encodes SEQ ID NO:259;(o) the first nucleotide sequence encodes SEQ ID NO:269, and the second nucleotide sequence encodes SEQ ID NO:277; or(p) the first nucleotide sequence encodes SEQ ID NO:287, and the second nucleotide sequence encodes SEQ ID NO:295.

[0020] In some embodiments,(a) the first nucleotide sequence comprises SEQ ID NO: 18 or a sequence that encodes SEQ ID NO: 17 and has at least 90% sequence identity to SEQ ID NO: 18, and the second nucleotide sequence comprises SEQ ID NO:26 or a sequence that encodes SEQ ID NO:25 andhas at least 90% sequence identity to SEQ ID NO:26;(b) the first nucleotide sequence comprises SEQ ID NO:36 or a sequence that encodes SEQ ID NO:35 and has at least 90% sequence identity to SEQ ID NO:36, and the second nucleotide sequence comprises SEQ ID NO:44 or a sequence that encodes SEQ ID NO:43 and has at least 90% sequence identity to SEQ ID NO: 44;(c) the first nucleotide sequence comprises SEQ ID NO:54 or a sequence that encodes SEQ ID NO:53 and has at least 90% sequence identity to SEQ ID NO:54, and the second nucleotide sequence comprises SEQ ID NO:62 or a sequence that encodes SEQ ID NO:61 and has at least 90% sequence identity to SEQ ID NO: 62;(d) the first nucleotide sequence comprises SEQ ID NO:72 or a sequence that encodes SEQ ID NO:71 and has at least 90% sequence identity to SEQ ID NO:72, and the second nucleotide sequence comprises SEQ ID NO:80 or a sequence that encodes SEQ ID NO:79 and has at least 90% sequence identity to SEQ ID NO: 80;(e) the first nucleotide sequence comprises SEQ ID NO:90 or a sequence that encodes SEQ ID NO:89 and has at least 90% sequence identity to SEQ ID NO:90, and the second nucleotide sequence comprises SEQ ID NO:98 or a sequence that encodes SEQ ID NO:97 and has at least 90% sequence identity to SEQ ID NO:98;(f) the first nucleotide sequence comprises SEQ ID NO: 108 or a sequence that encodes SEQ ID NO: 107 and has at least 90% sequence identity to SEQ ID NO: 108, and the second nucleotide sequence comprises SEQ ID NO: 116 or a sequence that encodes SEQ ID NO: 115 and has at least 90% sequence identity to SEQ ID NO: 116;(g) the first nucleotide sequence comprises SEQ ID NO: 126 or a sequence that encodes SEQ ID NO: 125 and has at least 90% sequence identity to SEQ ID NO: 126, and the second nucleotide sequence comprises SEQ ID NO: 134 or a sequence that encodes SEQ ID NO: 113 and has at least 90% sequence identity to SEQ ID NO: 134;(h) the first nucleotide sequence comprises SEQ ID NO: 144 or a sequence that encodes SEQ ID NO: 143 and has at least 90% sequence identity to SEQ ID NO: 144, and the second nucleotide sequence comprises SEQ ID NO: 152 or a sequence that encodes SEQ ID NO: 151 and has at least 90% sequence identity to SEQ ID NO: 152;(i) the first nucleotide sequence comprises SEQ ID NO: 162 or a sequence that encodes SEQ ID NO: 161 and has at least 90% sequence identity to SEQ ID NO: 162, and the second nucleotide sequence comprises SEQ ID NO: 170 or a sequence that encodes SEQ ID NO: 169 and has at least 90% sequence identity to SEQ ID NO: 170;(j) the first nucleotide sequence comprises SEQ ID NO: 180 or a sequence thatencodes SEQ ID NO: 179 and has at least 90% sequence identity to SEQ ID NO: 180, and the second nucleotide sequence comprises SEQ ID NO: 188 or a sequence that encodes SEQ ID NO: 187 and has at least 90% sequence identity to SEQ ID NO: 188;(k) the first nucleotide sequence comprises SEQ ID NO: 198 or a sequence that encodes SEQ ID NO: 197 and has at least 90% sequence identity to SEQ ID NO: 198, and the second nucleotide sequence comprises SEQ ID NO:206 or a sequence that encodes SEQ ID NO:205 and has at least 90% sequence identity to SEQ ID NO:206;(l) the first nucleotide sequence comprises SEQ ID NO:2165 or a sequence that encodes SEQ ID NO:215 and has at least 90% sequence identity to SEQ ID NO:216, and the second nucleotide sequence comprises SEQ ID NO:224 or a sequence that encodes SEQ ID NO:223 and has at least 90% sequence identity to SEQ ID NO:224;(m) the first nucleotide sequence comprises SEQ ID NO:234 or a sequence that encodes SEQ ID NO:233 and has at least 90% sequence identity to SEQ ID NO:234, and the second nucleotide sequence comprises SEQ ID NO:242 or a sequence that encodes SEQ ID NO:241 and has at least 90% sequence identity to SEQ ID NO:242;(n) the first nucleotide sequence comprises SEQ ID NO:252 or a sequence that encodes SEQ ID NO:251 and has at least 90% sequence identity to SEQ ID NO:252, and the second nucleotide sequence comprises SEQ ID NO:260 or a sequence that encodes SEQ ID NO:259 and has at least 90% sequence identity to SEQ ID NO:260;(o) the first nucleotide sequence comprises SEQ ID NO:270 or a sequence that encodes SEQ ID NO:269 and has at least 90% sequence identity to SEQ ID NO:270, and the second nucleotide sequence comprises SEQ ID NO:278 or a sequence that encodes SEQ ID NO:277 and has at least 90% sequence identity to SEQ ID NO:278; or(p) the first nucleotide sequence comprises SEQ ID NO:288 or a sequence that encodes SEQ ID NO:287 and has at least 90% sequence identity to SEQ ID NO:288, and the second nucleotide sequence comprises SEQ ID NO:296 or a sequence that encodes SEQ ID NO:295 and has at least 90% sequence identity to SEQ ID NO:296.

[0021] In some embodiments, the nucleotide sequence encoding the a chain and the nucleotide sequence encoding the P chain are present on a single nucleic acid or vector. In some embodiments, the nucleotide sequence encoding the a chain and the nucleotide sequence encoding the P chain are present on a single nucleic acid or vector and expressed from a single promoter. In some embodiments, the nucleotide sequence encoding the a chain and the nucleotide sequence encoding the P chain are present on a single nucleic acid or vector, separated by a peptide sequence that causes ribosome skipping or self-cleavage, and expressedfrom a single promoter. The peptide that causes ribosome skipping can be, but is not limited to, a 2A peptide. The 2A peptide can be, but is not limited to, a P2A peptide. In some embodiments, the P2A peptide comprises SEQ ID NO:301.

[0022] In some embodiments, the nucleotide sequence encodes the amino acid sequence of SEQ ID NO:27, 45, 63, 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, or 297.

[0023] In some embodiments, the nucleotide sequence comprises:(a) SEQ ID NO:28 or a sequence that encodes SEQ ID NO:27 and has at least 90% sequence identity to SEQ ID NO:28;(b) SEQ ID NO:46 or a sequence that encodes SEQ ID NO:45 and has at least 90% sequence identity to SEQ ID NO:46:(c) SEQ ID NO: 64 or a sequence that encodes SEQ ID NO: 63 and has at least 90% sequence identity to SEQ ID NO 64;(d) SEQ ID NO: 82 or a sequence that encodes SEQ ID NO: 81 and has at least 90% sequence identity to SEQ ID NO: 82;(e) SEQ ID NO: 100 or a sequence that encodes SEQ ID NO: 99 and has at least 90% sequence identity to SEQ ID NO: 100;(f) SEQ ID NO: 118 or a sequence that encodes SEQ ID NO: 117 and has at least 90% sequence identity to SEQ ID NO: 118;(g) SEQ ID NO: 136 or a sequence that encodes SEQ ID NO: 135 and has at least 90% sequence identity to SEQ ID NO: 136;(h) SEQ ID NO: 154 or a sequence that encodes SEQ ID NO: 153 and has at least 90% sequence identity to SEQ ID NO: 154;(i) SEQ ID NO: 172 or a sequence that encodes SEQ ID NO: 171 and has at least 90% sequence identity to SEQ ID NO: 172;(j) SEQ ID NO: 190 or a sequence that encodes SEQ ID NO: 189 and has at least 90% sequence identity to SEQ ID NO: 190;(k) SEQ ID NO:208 or a sequence that encodes SEQ ID NO:207 and has at least 90% sequence identity to SEQ ID NO:208;(l) SEQ ID NO:226 or a sequence that encodes SEQ ID NO:225 and has at least 90% sequence identity to SEQ ID NO: 226;(m) SEQ ID NO:244 or a sequence that encodes SEQ ID NO:243 and has at least 90% sequence identity to SEQ ID NO: 244;(n) SEQ ID NO:262 or a sequence that encodes SEQ ID NO:261 and has at least 90% sequence identity to SEQ ID NO: 262;(o) SEQ ID NO:280 or a sequence that encodes SEQ ID NO:279 and has at least 90% sequence identity to SEQ ID NO:280; or(p) SEQ ID NO: 298 or a sequence that encodes SEQ ID NO: 297 and has at least 90% sequence identity to SEQ ID NO:298.

[0024] Also provided herein are vectors comprising any of the polynucleotides provided herein. In some embodiments, the vector is a viral vector. Viral vectors include, but are not limited to, retroviral, adenoviral, and adeno-derived vectors, Retroviral vectors include, but are not limited to, lentivirus. In some embodiments, the viral vector is a lentiviral vector. Additional vectors include, but are not limited to, transposon-based systems, mini-circles, mRNA, and CRISPR constructs.

[0025] Also provided herein are engineered cells comprising any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein. In some embodiments, the anti-NPMlc TCR or antigen-binding fragment thereof is heterologous to the cell. Also provided herein are engineered cells comprising any of the nucleotide sequence provided herein or any of the vectors provided herein. In some embodiments, the nucleotide sequences encoding the anti- NPMlc TCR or antigen-binding fragment thereof are heterologous to the cell. The cell can be, but is not limited to, a T cell, a cell line T cell, a primary T cell, a donor T cell (z.e., a T cell derived from a subject that is not the patient), or an autologous T cell (e.g., a T cell derived from the subject to be treated with the engineered T cells).

[0026] Also provided herein are methods for producing an engineered cell that involve introducing any of the polynucleotides provided herein or any of the vectors provided herein into a cell to form the engineered cell. In some embodiments, the anti-NPMlc TCR or antigenbinding fragment thereof is heterologous to the cell. The cell can be, but is not limited to, a T cell, a cell line T cell, a primary T cell, a donor T cell (z.e., a T cell derived from a subject that is not the patient), or an autologous T cell (i.e., a T cell derived from the subject to be treated with the engineered T cells).

[0027] Also provided are soluble anti-NPMlc TCRs and anti-NPMlc TCR bispecific molecules. Anti-NPMlc TCR bispecific molecules comprise fusion proteins containing an antigen-binding domain from any of the described anti-NPMlc TCRs and a second antigenbinding domain that specifically binds a second antigen wherein the second antigen is not a NPMlc neoantigen. The second antigen can be, but is not limited to, a T cell antigen (e.g., CD3), a macrophage antigen, or a natural killer (NK) cell antigen. In some embodiments, the second antigen is an anti-CD3 binding protein. The anti-CD3 binding protein can be, but is not limited to, an anti-CD3 scFv.

[0028] In some embodiments, a soluble anti-NPMlc TCR or antigen binding fragment thereof is linked to or fused to an immune cell engager. The immune cell engager can be, but is not limited to, T cell engager (e.g., a CD3 binding domain (e.g., an anti-CD3 scFv)), a macrophage engager, or a NK cell engager. In some embodiments, a soluble anti-NPMlc TCR or antigen binding fragment thereof is linked to or fused to a Fc peptide, a modified Fc peptide, or a Tc receptor, or a modified Fc receptor.

[0029] Also provided herein are compositions comprising any of the provided anti-NPMlc TCRs or antigen-binding fragments thereof, any of the polynucleotides provided herein, any of the vectors provided herein, or any of the engineered cells provided herein. In some embodiments, the compositions also include a pharmaceutically acceptable excipient.

[0030] Also provided herein are methods of treatment comprising administering to a subject having or diagnosed with an NPMlc+cancer, any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein. Also provided herein are any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein, for use in the treatment of NPMlc+cancer in a subject. Also provided herein are uses of any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein in the manufacture of a medicament for the treatment of NPMlc+cancer in a subject. In some embodiments, the engineered T cells are engineered autologous T cells. In some embodiments, the NPMlc+cancer is AML.

[0031] In some embodiments, methods of treating a subject having or diagnosed with an NPMlc+cancer are described, the methods comprising administering to a subject any of the engineered T cells provided herein. In some embodiments, any of the engineered T cells described herein are for use in the treatment of NPMlc+cancer in a subject. In some embodiments, any of the engineered T cells provided herein can be used in the manufacture of a medicament for the treatment of NPMlc+cancer in a subject. In some embodiments, the engineered T cells are engineered autologous T cells. In some embodiments, the NPMlc+cancer is AML.

[0032] In some embodiments, methods of treating a subject having or diagnosed with an NPMlc+cancer are described, the methods comprising administering to a subject any of thesoluble anti-NPMlc TCRs (or antigen-binding fragments thereof) or anti-NPMlc TCR bispecific molecules provided herein. In some embodiments, any of the soluble anti-NPMlc TCRs (or antigen-binding fragments thereof) or anti-NPMlc TCR bispecific molecules described herein are for use in the treatment of NPMlc+cancer in a subject. In some embodiments, any of the soluble anti-NPMlc TCRs (or antigen-binding fragments thereof) or anti-NPMlc TCR bispecific molecules provided herein can be used in the manufacture of a medicament for the treatment of NPMlc+cancer in a subject.

[0033] In some embodiments, the subject is not eligible for standard of care transplant. In some embodiments, the subject is FLT3 positive. In some embodiments, the subject has or has been diagnosed with AML. In some embodiments, the subject has an NPMlc+cancer and one or more co-morbidities or risk factors. In some embodiments, the subject has AML and one or more co-morbidities or risk factors. In some embodiments, the subject has an NPMlc+cancer and is ineligible for high-intensity chemotherapy. In some embodiments, the subject has AML and is ineligible for high-intensity chemotherapy. In some embodiments, the subject has relapsed following high-intensity chemotherapy with or without hematopoietic stem cell transplant (HSCT). In some embodiments, the subject has not responded to, is not responding to, or has failed to achieve complete remission (is refractory to) high-intensity chemotherapy. In some embodiments, the subject has relapsed following a prior hypomethylating agent therapy. In some embodiments, the subject has not responded, is not responding to, or has failed to achieve complete remission (is refractory to) hypomethylating agent therapy. In some embodiments, the subject has relapsed or has not responded to at least two prior therapies. Relapse can occur following, e.g., complete remission (CR), CR with partial hematologic recovery (CRh), or CR with incomplete recovery (CRi). In some embodiments, the subject has relapsed or refractory AML. In some embodiments, the subject has AML and has relapsed following at least one prior therapy. In some embodiments, the subject has AML and has relapsed following at least two prior therapies. In some embodiments, the subject has AML and is refractory to at least one prior therapy. In some embodiments, the subject has AML and is refractory to at least two prior therapies.

[0034] In some embodiments, administration of the engineered cell or the composition induces or enhances death of cells associated with the malignant hematologic disorder, or induces or enhances a graft versus leukemia effect (GVL) in the subject.BRIEF DESCRIPTION OF THE FIGURES

[0035] FIG. 1. Graph illustrating activation of engineered T cells expressing TCR D inresponse to cells pulsed with CLAVEEVSL (SEQ ID NO:5) peptide.

[0036] FIG. 2. Graph illustrating target cell killing of cells pulsed with CLAVEEVSL (SEQ ID NO:5) peptide by engineered T cells expressing TCR D.

[0037] FIG. 3. Graph illustrating target cell killing of cells naturally expressing the CLAVEEVSL (SEQ ID NO:5) peptide by engineered T cells expressing TCR D.

[0038] FIG. 4. Graphs illustrating efficacy of engineered T cells compared to CAR T cells.

[0039] FIG. 5. Graphs illustrating cytotoxicity (top) and EC so (bottom) of engineered T cells compared to CAR T cells.

[0040] FIG. 6. Graph illustrating reactivity of TCR D against cysteinylated and non- cysteinylated target peptide.

[0041] FIG. 7. Graph illustrating recognition of CLAVEEVSL (SEQ ID NO:5) variants by TCR F. Peptide number corresponds to the peptides of Table 11. Each peptide was tested at, in order, 5 pg / mL, 0.16 pg / mL, 0.015 pg / mL, and 0 pg / mL.

[0042] FIG. 8. Graph illustrating recognition of CLAVEEVSL (SEQ ID NO: 5) variants by TCR D. Peptide number corresponds to the peptides of Table 11. Each peptide was tested at, in order, 5 pg / mL, 0.16 pg / mL, 0.015 pg / mL, and 0 pg / mL.DETAILED DESCRIPTION

[0043] Provided herein are anti-NPMlc TCRs, including recombinant anti-NPMlc TCRs, that bind or recognize a neoantigen associated with NPMlc. Among the provided embodiments are approaches useful in the treatment of such diseases and conditions and / or for targeting cell types, such as cancer cells or cells associated with expression of NPMlc. In some embodiments, the provided anti-NPMlc TCRs and antigen-binding fragments thereof, specifically bind or recognize a neoantigen having the sequence AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5). In some embodiments, the provided anti-NPMlc TCRs and antigen-binding fragments thereof, specifically bind or recognize a neoantigen having the sequence AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule. In some embodiments, the provided anti-NPMlc TCRs and antigenbinding fragments thereof, specifically bind or recognize a neoantigen having the sequence: AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an HLA- A*02:01, HLA-A*02:06, or HLA-A*02:03.

[0044] In some embodiments, the described anti -AIQDLCLAV (SEQ ID NO: 1) anti-NPMlc TCRs specifically bind a AIQDLCLAV (SEQ ID NO: 1) neoantigen when presented by HLA subtype A*02:01, HLA subtype A* 02: 06, or HLA subtype A* 02: 03 on a cell (e.g., a cancercell), do not specifically bind the AIQDLCLAV (SEQ ID NO: 1) neoantigen when bound to other HLA subtypes or other non-target antigens presented by HLA subtype A*02:01, HLA subtype A* 02: 06, or HLA subtype A* 02: 03.

[0045] In some embodiments, the described anti-CLAVEEVSL (SEQ ID NO: 5) anti- NPMlc TCRs specifically bind a CLAVEEVSL (SEQ ID NO: 5) neoantigen when presented by HLA subtype A*02:01, HLA subtype A*02:06, or HLA subtype A*02:03 on a cell e.g., a cancer cell), do not specifically bind the CLAVEEVSL (SEQ ID NO: 5) neoantigen when bound to other HLA subtypes or other non-target antigens presented by HLA subtype A*02:01, HLA subtype A* 02: 06, or HLA subtype A* 02: 03.

[0046] Also provided herein are nucleic acid molecules encoding the anti-NPMlc TCRs, engineered cells containing the anti-NPMlc TCRs, compositions containing the anti-NPMlc TCRs or cells, and methods of treatment or uses, such as therapeutic uses, involving administering such anti-NPMlc TCRs, engineered cells, or compositions, and uses of such anti-NPMlc TCRs, cells or compositions. In some aspects, engineered cells that express a provided anti-NPMlc TCR or antigen binding fragment thereof, exhibit cytotoxic activity against target cells expressing the NPMlc neoantigen, such as cancer cells. Also provided herein are methods for identifying an anti-NPMlc TCR targeting a neoantigen having the sequence AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5).

[0047] Cell therapies (including those involving the administration of cells engineered to express recombinant receptors or anti-NPMlc TCRs specific for a disease or condition of interest, such as a recombinant anti-NPMlc TCR and / or other recombinant antigen receptors), as well as other adoptive immune cell and adoptive T cell therapies can be effective in the treatment of diseases and disorders. T cells engineered to express the described anti-NPMlc TCRs (or antigen binding fragments thereof) recognize and bind to a target, e.g. , target antigen, such as peptide epitopes of AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5), within the subject, for example, based on the affinity of the antigen-binding domain of the anti- NPMlc TCR to its target antigen.

[0048] In some embodiments, a T cell engineered to express a heterologous anti-NPMlc TCR, (e.g, an anti-AIQDLCLAV TCR or an anti-CLAVEEVSL TCR), is further modified to knock out the endogenous TCR. Knock out of the endogenous TCR can include knock out of expression of the endogenous TRAC gene and knock out of expression of the endogenous TRBC gene. In some embodiments, knock out of the endogenous TCR (the endogenous TRAC gene and / or the endogenous TRBC gene) can be performed using CRISPR. Knockout of the endogenous TCR includes, but is not limited to, deleting all or a port of the target gene,inserting one or more stop codons into the target gene, disrupting the target gene, or disrupting the promotor or a splicing region of the target gene. Knock out of the endogenous TCR reduces or eliminates mispairing of the heterologous anti-NPMlc TCR (a and / or P chain) with the endogenous TCR (a and / or p chain).

[0049] In some aspects, the provided anti-AIQDLCLAV TCRs or anti-CLAVEEVSL TCRs have improved affinity to the NPMlc neoantigen. In some aspects, engineered T cells expressing any of the described anti-AIQDLCLAV or anti-CLAVEEVSL TCRs have improved activity against cells (e.g., cancer cells) expressing an NPM-lc neoantigen. In some aspects, engineered T cells expressing any of the described anti-AIQDLCLAV or anti- CLAVEEVSL TCRs have improved activity against cells (e.g., cancer cells) expressing an NPM-lc neoantigen at a low effector to target (E:T) ratio.

[0050] In some aspects, therapeutic approaches using the described anti-NPMlc TCRs, for example in adoptive cell therapy with engineered human T cells expressing the described anti- NPMlc TCRs, result target and kill cancer cells expressing an NPMlc neoantigen, leading to Graft vs. Leukemia (GVL) effect.

[0051] All publications, including patent documents, scientific articles and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. If a definition set forth herein is contrary to or otherwise inconsistent with a definition set forth in the patents, applications, published applications and other publications that are herein incorporated by reference, the definition set forth herein prevails over the definition that is incorporated herein by reference.

[0052] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.I. T CELL RECEPTORS TARGETING NPM1 NEOANTIGENS

[0053] Provided herein are anti-NPMlc TCRs, such as those that bind or recognize a peptide epitope associated with a NPMlc, such as a peptide neoantigen expressed on the surface of an immune cell, a cancer cell and / or a cell associated with a hematological ailment, in the context of an MHC molecule. In some embodiments, the described anti-NPMlc TCRs bind or recognize the neoantigen AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5), in the context of an MHC molecule. Such anti-NPMlc TCRs and antigen-binding fragments exhibit antigenic specificity for binding or recognizing such peptide epitopes. Also provided are (a) nucleic acids encoding the anti-NPMlc TCRs (or antigen binding domains thereof), (b)engineered cells expressing the anti-NPMlc TCRs (or antigen-binding domains thereof), (c) compositions comprising the anti-NPMlc TCRs, nucleic acids encoding the anti-NPMlc TCRs, or engineered cells expressing the anti-NPMlc TCRs, and (d) methods of treatment involving administering the described anti-NPMlc TCRs (or antigen-binding domains thereof), nucleic acids, engineered cells, or compositions. In some aspects, engineered cells that express a provided anti-NPMlc TCR or antigen-binding fragment thereof, exhibit cytotoxic activity against target cells expressing the NPMlc neoantigen, such as NPMlc+cancer cells.

[0054] In some embodiments, the described anti-NPMlc TCRs recognize a NPMlc neoantigen (e.g, AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5)) expressed by AML cancer cells.A. NPMlc

[0055] Nucleophosmin (NPM, also known as nucleolar phosphoprotein B23 or numatrin) is encoded in humans by the NPM1 gene. The NPM1 gene is up-regulated and mutated in certain tumor types, including leukemias, acute myelogenous (myeloid) leukemia (AML), and myelodysplastic syndrome. Mutations in NPM1 often consist of 4 bp insertions or duplications between nucleotides 960 and 961. They cause the replacement of the last seven amino acids (WQWRKSL) with 11 different residues and result in cytoplasmic localization of the protein (NPMlc).

[0056] NPMlc neoantigens include, but are not limited to: AIQDLCLAV (SEQ ID NO: 1) and CLAVEEVSL (SEQ ID NO: 5). The corresponding peptide sequence in the non-mutant NPM1 is AIQDLWQWRKSL (SEQ ID NO:2). The NPMlc neoantigens can be presented in the context of Class I MHC molecule, e.g., HLA-A*02:01, HLA-A*02:06, or HLA-A*02:03. The NPMlc neoantigen AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) is typically expressed by leukemia cells (e.g., AML cells), and is not typically expressed by nonleukemia cells.

[0057] In some aspects, the provided anti-NPMlc TCRs, or antigen-binding fragments thereof, recognize or specifically bind to an immunogenic epitope of NPMlc. In some examples, the provided anti-NPMlc TCRs, or antigen-binding fragments thereof, specifically bind to or recognize the NPMlc neoantigen AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5), but do not specifically bind to the corresponding normal NPM1 peptide, AIQDLWQWRKSL (SEQ ID NO:2). In some aspects, the anti-NPMlc TCR (or antigenbinding fragments thereof) recognizes or binds an NPMlc neoantigen (e.g., AIQDLCLAV(SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5)) in the context of an MHC molecule, such as an MHC Class I molecule. In some aspects, the MHC Class I molecule is an HLA-A2 molecule, including any one or more subtypes thereof, e.g., HLA-A*02:01, *02:06, or *02:03.

[0058] In some embodiments, the anti-NPMlc TCR or soluble anti-NPMlc TCR, or antigenbinding fragment thereof, is isolated or purified. In particular embodiments, any of the provided anti-NPMlc TCRs, or antigen-binding fragments thereof, are recombinant. In some aspects, the anti-NPMlc TCR, or antigen-binding fragment thereof, is human. In some aspects, the anti- NPMlc TCR, or antigen-binding fragment thereof, is a single chain. In other embodiments, the anti-NPMlc TCR contains two chains. In some embodiments, the anti-NPMlc TCR, or antigen-binding fragment thereof, is expressed on the surface of a cell (e.g., a T cell such as a T cell designed to lack expression of endogenous TCRs). In some embodiments, a soluble anti- NPMlc TCR is secreted from a cell.

[0059] In some aspects, the provided anti-NPMlc TCRs, or antigen-binding fragment thereof, have one or more specified functional features, such as binding properties, including binding to particular epitopes, and / or particular binding affinities, for example, as described herein. In some aspects, engineered cells, such as engineered T cells, expressing the provided anti-NPMlc TCRs (or antigen-binding fragment thereof) have one or more specified functional features, such as binding properties, including binding to particular epitopes, particular binding affinities, activation or stimulation of cell signaling, such as T cell signaling or TCR signaling, secretion of cytokines, and / or killing of target cells expressing or presenting the antigen, for example, as described herein.

[0060] Provided are anti-NPMlc TCRs, or antigen-binding fragment thereof, that specifically bind an NPMlc neoantigen. A TCR is a molecule that contains an a chain comprising a Va region and a P chain comprising a VP region (also known as TCRa and TCRP, respectively) or a y chain comprising a Vy region and a 5 chain comprising a V5 region (also known as TCRy and TCRS, respectively), or antigen-binding portions thereof, which is capable of specifically binding to an antigen, e.g., a peptide antigen or peptide epitope bound to an MHC molecule. In some embodiments, the anti-NPMlc TCR is in the aP form (e.g., is an aP TCR). In some embodiments, the anti-NPMlc TCR is in the yS form (e.g, is an yS TCR). Typically, TCRs that exist in aP or yS forms are generally structurally similar, but T cells expressing them may have distinct anatomical locations or functions. A TCR can be found on the surface of a cell or in soluble form. Generally, a TCR is found on the surface of T cells where it is generally responsible for recognizing antigens, such as peptides bound to MHCmolecules.

[0061] In some embodiments, an anti-NPMlc TCR provided herein can be an intact or full- length TCR, such as a TCR containing a full length a chain and a full length P chain, or a TCR containing a full length y chain and a full length 5 chain. In some embodiments, an antigenbinding portion of a TCR provided herein can be less than a full-length TCR provided that it binds to a specific peptide bound in an MHC molecule. In some embodiments, an antigenbinding portion or fragment of a TCR can contain only a portion of the structural domains of a full-length or intact TCR, but yet is able to bind the peptide epitope, such as MHC -peptide complex, to which the full-length TCR binds. In some embodiments, an antigen-binding portion contains the variable domains of a TCR, such as a Va region and a VP region of a TCR, or a Vy region and a V5 region of a TCR provided herein provided that that antigen-binding portion is sufficient to form a binding site for binding to a specific MHC -peptide complex.

[0062] The variable domains of the TCR contain complementarity-determining regions (CDRs; CDR-1, CDR-2, and CDR-3), which contribute to antigen recognition and binding capabilities and specificity of the peptide, the MHC molecule, and / or MHC-peptide complex. In some embodiments, one or more CDRs of a TCR form all or substantially all of the antigenbinding site of a given TCR molecule. The various CDRs within a variable region of a TCR chain generally are separated by framework regions (FRs), which generally display less variability among TCRs as compared to the CDRs. CDR-3 is the main CDR responsible for antigen binding or specificity, or is the most important among the three CDRs on a given TCR variable region for antigen recognition, and / or for interaction with the processed peptide portion of the peptide-MHC complex. In some contexts, CDR-1 of the a chain interacts with the N-terminal part of certain antigenic peptides. In some contexts, CDR-1 of the P chain interacts with the C-terminal part of the peptide. In some contexts, CDR-2 contributes most strongly to or is the primary CDR responsible for the interaction with or recognition of the MHC portion of the MHC-peptide complex.

[0063] In some embodiments, the a chain and / or the P chain of a TCR, or the y chain and / or the 5 chain of a TCR, also can contain one or more of: a constant domain, a transmembrane domain and / or a short cytoplasmic tail. In some aspects, each chain (e.g., a or P) of the TCR contains one N-terminal immunoglobulin variable domain, one immunoglobulin constant domain, a transmembrane region, and a short cytoplasmic tail at the C-terminal end. In some embodiments, a TCR, for example via the cytoplasmic tail, is associated with invariant proteins of the CD3 complex involved in mediating signal transduction. In some embodiments, the structure allows the TCR to associate with other molecules like CD3 and subunits thereof. Forexample, a TCR containing constant domains with a transmembrane region may anchor the protein in the cell membrane and associate with invariant subunits of the CD3 signaling apparatus or complex. The intracellular tails of CD3 signaling subunits (e.g., CD3y, CD35, CD3s and CD3(^ chains) contain one or more immunoreceptor tyrosine-based activation motif or IT AM and generally are involved in the signaling capacity of the TCR complex.

[0064] The various domains or regions of a TCR can be identified. In some embodiments, the exact locus of a domain or region can vary depending on the particular structural or homology modeling or other features used to describe a particular domain. It is understood that reference to amino acids, including to a specific sequence set forth as a SEQ ID NO: used to describe domain organization of a TCR are for illustrative purposes and are not meant to limit the scope of the embodiments provided. In some embodiments, the specific domain (e.g., variable or constant) can be several amino acids (such as one, two, three or four) longer or shorter. In some aspects, residues of a TCR are known or can be identified according to the INTERNATIONAL IMMUNOGENETICS INFORMATION SYSTEM® (IMGT) numbering system (Lefranc et al. (2003) Developmental and Comparative Immunology, 27(l);55-77; and The T Cell Factsbook 2nd Edition, Lefranc and LeFranc Academic Press 2001). Using this system, CDR-1 sequences within a TCR Va region and / or VP region in some cases correspond to the amino acids present between residue numbers 27-38, inclusive, CDR-2 sequences within a TCR Va region and / or VP region in some cases correspond to the amino acids present between residue numbers 56-65, inclusive, and CDR-3 sequences within a TCR Va region and / or VP region in some cases correspond to the amino acids present between residue numbers 105-117, inclusive.

[0065] In some embodiments, provided are anti-NPMlc TCRs or antigen-binding fragments thereof provided that specifically bind to or recognize the NPMlc neoantigen AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5), in the context of an MHC molecule. In some embodiments, the provided anti-NPMlc TCRs or antigen-binding fragments thereof do not recognize or specifically bind to the NPM1 peptide AIQDLWQWRKSL (SEQ ID NO:2). In some embodiments, the provided anti-NPMlc TCRs or antigen-binding fragments thereof specifically recognize or bind to AIQDLCLAV (SEQ ID NO: 1), and do not specifically recognize or bind to a AIQDLWQWRKSL (SEQ ID NO:2). In some embodiments, the provided anti-NPMlc TCRs or antigen-binding fragments thereof bind to AIQDLCLAV (SEQ ID NO: 1) with increased affinity relative to binding to AIQDLWQWRKSL (SEQ ID NO:2). In some embodiments, the provided anti-NPMlc TCRs or antigen-binding fragments thereof specifically recognize or bind to CLAVEEVSL (SEQ ID NO: 5), and do not specificallyrecognize or bind to a AIQDLWQWRKSL (SEQ ID NO:2). In some embodiments, the provided anti-NPMlc TCRs or antigen-binding fragments thereof bind to CLAVEEVSL (SEQ ID NO: 5) with increased affinity relative to binding to AIQDLWQWRKSL (SEQ ID NO:2).

[0066] In some aspects, the provided anti-NPMlc TCRs or antigen-binding fragments thereof specifically bind to or recognize AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) that is complexed with an MHC molecule of a particular HLA type, such as HLA- A*02:01, HLA-A*02:06, or HLA-A*02:03.

[0067] In some embodiments, an anti-NPMlc TCR provided herein is a full-length TCR. In some embodiments, an anti-NPMlc TCR provided herein is a dimeric TCR (dTCR). In some embodiments, anti-NPMlc TCR provided herein is a single-chain TCR (scTCR). An anti- NPMlc TCR as provided herein may be cell-bound or in soluble form. In some embodiments, an anti-NPMlc TCR as provided herein is in cell-bound form expressed on the surface of a cell (e.g., a T cell such as a T cell designed to lack expression of endogenous TCRs).

[0068] In some embodiments, an anti-NPMlc TCR provided herein is a scTCR, which is a single amino acid strand containing an a chain and a P chain that is able to bind to MHC- peptide complexes. Typically, a scTCR can be generated as described elsewhere, see, e.g., WO 96 / 13593, WO 96 / 18105, WO99 / 18129, WO 04 / 033685, W02006 / 037960, WO2011 / 044186; U.S. Patent No. 7,569,664; and Schlueter, C. J. et al. J. Mol. Biol. 256, 859 (1996).B. Exemplary Variable Domains

[0069] Provided herein are anti-NPMlc TCRs or antigen-binding fragments thereof that specifically recognize or bind the neoantigen AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5), in the context of an MHC molecule. Provided are exemplary sequences (e.g., CDRs, Va and / or VP, or Vy and / or V5, and constant region sequences) of AIQDLCLAV (SEQ ID NO: l)-specific or CLAVEEVSL (SEQ ID NO: 5)-specific TCRs.

[0070] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein specifically binds to or recognizes the neoantigen AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) presented on the surface of leukemia cells. In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein specifically binds to or recognizes the neoantigen AIQDLCLAV (SEQ ID NO: 1) presented on the surface of AML cells. In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein specifically binds to or recognizes the neoantigen AIQDLCLAV (SEQ ID NO: 1) presented on the surface of myelodysplastic syndrome cells. In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided hereinspecifically binds to or recognizes the neoantigen CLAVEEVSL (SEQ ID NO: 5) presented on the surface of AML cells. In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein specifically binds to or recognizes the neoantigen CLAVEEVSL (SEQ ID NO: 5) presented on the surface of myelodysplastic syndrome cells.

[0071] In some aspects, cytotoxic activity of T cells expressing anti-AIQDLCLAV or anti- CLAVEEVSL TCRs, is stimulated upon contact of the T cells with target cells presenting or expressing the NPMlc (AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5)) neoantigen.

[0072] In some embodiments, the anti-NPMlc TCRs or antigen-binding fragment thereof contain any of the Va region and VP region, or Vy region and V5 region, sequences as described, or a sufficient antigen-binding portion of such sequences. In some embodiments, the described anti-NPMlc TCRs or antigen-binding fragments thereof contain Va and VP region, or Vy and V5 region, sequences or sufficient antigen-binding portions thereof that contain CDR-3 sequences as described herein. In some embodiments, the described anti- NPMlc TCRs or antigen-binding fragments thereof contain Va and VP region, or Vy and V5 region, sequences or sufficient antigen-binding portions thereof that contain CDR-1, a CDR-2 and CDR-3 sequences as described herein. Also, among the provided anti-NPMlc TCRs are those having sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such a sequence.

[0073] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein contains a Va or Vy region containing a CDR-3 comprising an amino acid sequence set forth in any of SEQ ID NOs: 13, 31, 49, 67, 85, 103, 121, 139, 157, 175, 193, 211, 229, 247, 265, and 283, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some aspects, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein contains a Va or Vy region containing a CDR-3 contained within the amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical with such a sequence.

[0074] In some embodiments, the Va or Vy region contains a CDR-1 comprising an amino acid sequence set forth in any of SEQ ID NOs: 11, 29, 47, 65, 83, 101, 119, 137, 155, 173, 191, 209, 227, 245, 263, and 281, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some aspects, the Va or Vy region contains a CDR-1 contained within the amino acid sequence set forth in any of SEQ IDNOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some embodiments, the Va or Vy region contains a CDR-2 comprising an amino acid sequence set forth in any of SEQ ID NOs: 12, 30, 48, 66, 84, 102, 120, 138, 156, 174, 192, 210, 228, 246, 264, and 282, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some embodiments, the Va or Vy region contains a CDR-2 contained within the amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence.

[0075] In some instances, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein contains a VP or V5 region containing a CDR-3 comprising an amino acid sequence set forth in any of SEQ ID NOs:21, 39, 57, 75, 93, 111, 129, 147, 165, 183, 201, 219, 237, 255, 273, and 291, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein contains a VP or V5 region containing a CDR-3 contained within the amino acid sequence set forth in any of SEQ ID NOs:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical with such a sequence.

[0076] In some instances, the VP or V5 region contains a CDR-1 comprising an amino acid sequence set forth in any of SEQ ID NOs: 19, 37, 55, 73, 91, 109, 127, 145, 163, 181, 199, 217, 235, 253, 271, and 289, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some aspects, the VP or V5 region contains a CDR-1 contained within the amino acid sequence set forth in any of SEQ ID NOs:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some embodiments, the VP or V5 region contains a CDR-2 comprising an amino acid sequence set forth in any of SEQ ID NOs:20, 38, 56, 74, 92, 110, 128, 146, 164, 182, 200, 218, 236, 254, 272, and 290, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence. In some embodiments, the VP or V5 region contains a CDR-2 contained within the amino acid sequence set forth in any of SEQ ID NOs:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,or 99% identity with such a sequence.

[0077] In some embodiments, the Va or Vy region contains the amino acid sequence set forth in any of SEQ ID NOs: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, and 284, or a sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto; and the VP or V5 region contains the amino acid sequence set forth in any of SEQ ID NOs:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, and 292, or a sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.

[0078] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule can include an a chain (or y chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 11, 29, 47, and 65 (or a variant thereof with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 12, 30, 48, and 66 (or a variant thereof with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NOs: 13, 31, and 49 (or a variant thereof with one or two amino acid modifications) and a P chain (or 5 chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 19, 37, and 55 (or a variant thereof with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO:20, 38, and 56 (or a variant thereof with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO:21, 39, and 57 (or a variant thereof with one or two amino acid modifications). Anti-NPMlc TCRs having these CDRs and the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule include, TCR A, TCR B, or TCR C (see Table 1).

[0079] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include an a chain (or y chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO:65, 83, 101, 119, 137, 155, 173, 191, 209, 227, 245, 263, or 281 (or a variant thereof with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 66, 84, 102, 120, 138, 156, 174, 192, 210, 228, 246, 264, or 282 (or a variant thereof with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NOs: 67, 85, 103, 121, 139, 157, 175, 193, 211, 229, 247, 265, or 283 (or a variant thereof with one or two amino acid modifications) and a P chain (or 5 chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO:73, 91, 109, 127, 145, 163, 181, 199, 217, 235, 253, 271, or 289 (or avariant thereof with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 74, 92, 110, 128, 146, 164, 182, 200, 218, 236, 254, 272, or 290 (or a variant thereof with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO:75, 93, 111, 129, 147, 165, 183, 201, 219, 237, 255, 273, or 291 (or a variant thereof with one or two amino acid modifications). Anti-NPMlc TCRs having these CDRs and the ability to specifically bind CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule include TCR D, TCR E, TCR F, TCR G, TCR H, TCR I, TCR J, TCR K, TCR L, TCR M, TCR N, TCR O, and TCR P (see Table 1).

[0080] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind an NPMlc neoantigen in the context of an MHC molecule and having an a chain (or y chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 11, 29, 47, 65, 83, 101, 119, 137, 155, 173, 191, 209, 227, 245, 263, or 281 (or a variant thereof with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 12, 30, 48, 66, 84, 102, 120, 138, 156, 174, 192, 210, 228, 246, 264, or 282 (or a variant thereof with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 13, 31, 49, 67, 85, 103, 121, 139, 157, 175, 193, 211, 229, 247, 265, or 283 (or a variant thereof with one or two amino acid modifications) and a P chain (or 5 chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 19; 37, 55, 73, 91, 109, 127, 145, 163, 181, 199, 217, 235, 253, 271, or 289 (or a variant thereof with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO:20, 38, 56, 56, 74, 92, 110, 128, 146, 164, 182, 200, 218, 236, 254, 272, or 290 (or a variant thereof with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO:21, 39, 57, 75, 93, 111, 129, 147, 165, 183, 201, 219, 237, 255, 273, or 291 (or a variant thereof with one or two amino acid modifications) can include any appropriate framework regions. For example, such an anti-NPMlc TCR or antigen binding fragment thereof can include an a chain that includes a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 that is upstream of the corresponding CDR1 amino acid sequence of SEQ ID NO: 11, 29, 47, 65, 83, 101, 119, 137, 155, 173, 191, 209, 227, 245, 263, or 281 (or a variant thereof with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 that is between the corresponding CDR1 and CDR2 amino acid sequences of SEQ ID NOs: 11 and 12, 29 and 30, 47 and 48, 65 and 66, 83and 84, 101 and 102, 119 and 120, 137 and 138, 155 and 156, 173 and 174, 191 and 192, 209 and 210, 227 and 228, 245 and 246, 263 and 264, or 281 and 282 (or a variant thereof with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the entire amino acid sequence set forth in SEQ ID NO: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 that is between the corresponding CDR2 and CDR 3 amino acid sequences of SEQ ID NOs: 12 and 13, 30 and 31, 48 and 49, 66 and 67, 84 and 85, 102 and 103, 120 and 121, 138 and 139, 156 and 157, 174 and 175, 192 and 193, 210 and 211, 228 and 229, 246 and 247, 264 and 264, or 282 and 283 (or a variant thereof with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 14, 32, 50, 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 that is downstream of the corresponding CDR3 amino acid sequence of SEQ ID NO: 13, 31, 49, 67, 85, 103, 121, 139, 157, 175, 193, 211, 229, 247, 265, or 283 (or a variant thereof with one, two, three, four, or five amino acid modifications) and a P chain that includes a framework region 1 having the entire amino acid sequence set forth in SEQ ID NO:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292 that is upstream of the corresponding CDR1 amino acid sequence of SEQ ID NO: 19, 37, 55, 73, 91, 109, 127, 145, 163, 181, 199, 217, 235,253, 271, or 289 (or a variant thereof with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the entire amino acid sequence set forth in SEQ ID NO:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292 that is between the corresponding CDR1 and CDR2 amino acid sequences of SEQ ID NOs: 19 and 20, 37 and 38, 55 and 56, 73 and 74, 91 and 92, 109 and 110, 127 and 128, 145 and 146, 163 and 164, 181 and 182, 199 and 200, 217 and 218, 235 and 236, 253 and254, 271 and 272, or 289 and 290 (or a variant thereof with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the entire amino acid sequence set forth in SEQ ID NO:22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292 that is between the corresponding CDR2 and CDR 3 amino acid sequences of SEQ ID NOs:20 and 21, 38 and 39, 56 and 57, 74 and 75, 92 and 93, 110 and 111, 128 and 129, 146 and 147, 164 and 165, 182 and 183, 200 and 201, 218 and 219, 236 and 237, 254 and 255, 272 and 273, or 290 and 291 (or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the entire amino acid sequence set forth in SEQ ID NO: 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292 that is downstream of the corresponding CDR3 amino acid sequence of SEQ ID NO:21, 39, 57, 75, 93, 111, 129, 147,165, 183, 201, 219, 237, 255, 273, or 291 (or a variant of that sequence with one, two, three, four, or five amino acid modifications).

[0081] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule can include an a chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 14, 32, or 50 and a P chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:22, 40, or 58. For example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 14, 32, or 50 and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:22, 40, or 58. In some embodiments, an anti- NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain and a P chain that include amino acid sequences having 100 percent identity to the amino acid sequence set forth in (a) SEQ ID NOs: 14 and 22; (b) SEQ ID NOs:32 and 40; or (c) SEQ ID NOs: 50 and 58.

[0082] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule can include (a) an a chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence of a Va domain of any one of TCRs A, B, and C (i.e., as set forth in SEQ ID NOs: 14, 32, and 50), provided that the a chain includes the corresponding CDR 1, CDR2, and CRD3 amino acid sequences of the TCR a chain (e.g., SEQ ID NOs: 29, 30, and 31 for TCR B) and (b) a P chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence of a VP domain of the corresponding TCR (i.e., as set forth in SEQ ID NOs:22, 40, and 58) provided that the P chain includes the corresponding CDR 1, CDR2, and CRD3 amino acid sequences of the TCR P chain (e.g., SEQ ID NOs:37, 38, and 39 for TCR B). For example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include (a) an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:32, provided that the a chain includes the amino acid sequences set forth in SEQ ID NOs:29, 30, and 3 land (b) a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 40, provided that the P chain includes theamino acid sequences set forth in SEQ ID NOs:37, 38, and 39.

[0083] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include an a chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 and a P chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292. For example, an anti- NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292. In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain and a P chain that include amino acid sequences having 100 percent identity to the amino acid sequence set forth (d) SEQ ID NO: 68 and 76; (e) SEQ ID NO:86 and 94; (f) SEQ ID NO: 104 and 112; (g) SEQ ID NO: 122 and 130; (h) SEQ ID NO: 140 and 148; (i) SEQ ID NO: 158 and 166; (j) SEQ ID NO: 176 and 184; (k) SEQ ID NO: 194 and 202; (1) SEQ ID NO:212 and 220; (m) SEQ ID NO:230 and 238; (n) SEQ ID NO:248 and 256; (o) SEQ ID NO:266 and 274; or (p) SEQ ID NO:284 and 292.

[0084] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include (a) an a chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence of a Va domain of any one of TCRs D through P (z.e., as set forth in SEQ ID NOs: 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284), provided that the a chain includes the corresponding CDR 1, CDR2, and CRD3 amino acid sequences of the TCR a chain (e.g., SEQ ID NOs; 65, 66, and 67 for TCR D; SEQ ID NOs: 83, 84, and 85 for TCR E; or SEQ ID NOs: 101, 102, and 103 for TCR F) and (b) a P chain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence of a VP domain of the corresponding TCR (z.e., as set forth in SEQ ID NOs:76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292) provided that the P chain includes the corresponding CDR 1, CDR2, and CRD3 amino acid sequences of the TCR P chain (e.g., SEQ ID NOs:73, 74, and 75 for TCR D; SEQ ID NOs:91, 92, and 93 for TCR E;and SEQ ID NOs: 109, 110, and 111 for TCR F). For example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include (a) an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 68, provided that the a chain includes the amino acid sequences set forth in SEQ ID NOs: 65, 66, and 67 and (b) a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:76, provided that the P chain includes the amino acid sequences set forth in SEQ ID NOs:73, 74, and 75.

[0085] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule can include the CDR sequences of TCR A, B or C and (a) an a chain having the amino acid sequence set forth in SEQ ID NO: 14, 32, or 50 or the amino acid set forth in SEQ ID NO: 14, with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions) and (b) a corresponding p chain (see Table 1) having the amino acid sequence set forth in SEQ ID NO:22, 40, or 58 or the amino acid set forth in SEQ ID NO:22, 40, or 58 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, an anti- NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule, can include (a) an a chain having the amino acid sequence set forth in SEQ ID NO:32 with zero, one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the a chain includes the CDR amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, and (b) a P chain having the amino acid sequence set forth in SEQ ID NO: 40 with zero, one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the P chain includes the CDR amino acid sequences set forth in SEQ ID NOs:37, 38, or 39.

[0086] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include the CDR sequences any of one of TCRs D-P and (a) a corresponding a chain (see Table 1) having the amino acid sequence set forth in SEQ ID NO: 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284 or the amino acid set forth in SEQ ID NO: 68, 86, 104, 122, 140, 158, 176, 194, 212, 230, 248, 266, or 284, with one, two,three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions) and (b) a corresponding p chain (see Table 1) having the amino acid sequence set forth in SEQ ID NO:76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292or the amino acid set forth in SEQ ID NO:76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, or 292 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule, can include (a) an a chain having the amino acid sequence set forth in SEQ ID NO: 68 with zero, one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the a chain includes the CDR amino acid sequences set forth in SEQ ID NOs:65, 66, and 67, and (b) a P chain having the amino acid sequence set forth in SEQ ID NO:76 with zero, one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the P chain includes the CDR amino acid sequences set forth in SEQ ID NOs:73, 73, and 74.

[0087] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind to AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include an a chain (or y chain) and a P chain (or 5 chain), having the CDR sequences (a or y chain CDR-1, CDR-2 and CDR-3 and P of 5 chain CDR-1, CDR-2, and CDR-3) as set forth in any of the anti-NPMlc TCRs in Table 1. In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include an a chain (or y chain) and a P chain (or 5 chain) having the CDR sequences (a or y chain CDR-1, CDR- 2 and CDR-3 and P of 5 chain CDR-1, CDR-2, and CDR-3) as set forth in any of the anti- NPMlc TCRs in Table 1, wherein any one or more of the CDRs can independently be a variant CDR having one or two amino acid modifications from the CDR sequence listed of Table 1.

[0088] Exemplary anti-NPMlc TCRs having these CDRs and the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule include TCR A, TCR B, and TCR C. As an example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule can include an a chain (or y chain) having a CDR-1 having the amino acidsequence set forth in SEQ ID NO:29 (or a variant of SEQ ID NO:29 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO:30 (or a variant of SEQ ID NO:30 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO:31 (or a variant of SEQ ID NO:31 with one or two amino acid modifications) and a P chain (or 5 chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO:37 (or a variant of SEQ ID NO:37 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO:38 (or a variant of SEQ ID NO:38 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO:39 (or a variant of SEQ ID NO:39 with one or two amino acid modifications).

[0089] Exemplary anti-NPMlc TCRs having these CDRs and the ability to specifically bind CLAVEEVSL (SEQ ID NO:5) in the context of an MHC molecule include TCRs D-P. As an example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO:5) in the context of an MHC molecule can include an a chain (or y chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 65 (or a variant of SEQ ID NO: 65 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 66 (or a variant of SEQ ID NO: 66 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 67 (or a variant of SEQ ID NO: 67 with one or two amino acid modifications) and a P chain (or 5 chain) having a CDR-1 having the amino acid sequence set forth in SEQ ID NO: 73 (or a variant of SEQ ID NO: 73 with one or two amino acid modifications), a CDR-2 having the amino acid sequence set forth in SEQ ID NO: 74 (or a variant of SEQ ID NO: 74 with one or two amino acid modifications), and a CDR-3 having the amino acid sequence set forth in SEQ ID NO: 75 (or a variant of SEQ ID NO: 75 with one or two amino acid modifications).

[0090] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule and having an a chain (or y chain) and a P chain (or 5 chain), having the CDR sequences (a or y chain CDR-1, CDR-2 and CDR-3 and P of 5 chain CDR-1, CDR-2, and CDR-3) as set forth in any of the anti-NPMlc TCRs in Table 1 can include any appropriate framework regions. In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5) in the context of an MHC molecule and having an a chain (or y chain) and a P chain (or 5 chain),having the CDR sequences (a or y chain CDR-1, CDR-2 and CDR-3 and P of 5 chain CDR-1, CDR-2, and CDR-3) as set forth in any of the anti-NPMlc TCRs in Table 1, wherein any one or more of the CDRs can independently be a variant CDR having one or two amino acid modifications from the CDR sequence listed of Table 1, can include any appropriate framework regions (z.e., a chain and P chain framework 1, 2, 3, and 4 regions). An a chain framework region 1 can have the amino acid sequence of any of the a variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is upstream of the corresponding CDR-1 sequence, or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications. An a chain framework region 2 can have the amino acid sequence of any of the a variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is between the corresponding CDR-1 and CDR-2 sequences, or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications. An a chain framework region 3 can have the amino acid sequence of any of the a variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is between the corresponding CDR-2 and CDR- 3 sequences, or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications. An a chain framework region 4 can have the amino acid sequence of any of the a variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is downstream of the corresponding CDR-3 sequence, or a variant of that sequence with one, two, three, four, or five amino acid modifications. A P chain framework region 1 can have the amino acid sequence of any of the P variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is upstream of the corresponding CDR-1 sequence, or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications. A P chain framework region 2 can have the amino acid sequence of any of the P variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is between the corresponding CDR-1 and CDR-2 sequences, or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications. A P chain framework region 3 can have the amino acid sequence of any of the P variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is between the corresponding CDR-2 and CDR-3 sequences, or a variant of that sequence with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications. A P chain framework region 4 can have the amino acid sequence of any of the P variable (V) region amino acid sequences of any of the anti-NPMlc TCRs set forth in Table 1 that is downstream of thecorresponding CDR-3 sequence, or a variant of that sequence with one, two, three, four, or five amino acid modifications. An anti-NPMlc TCR or antigen binding fragment thereof can have the a chain framework regions 1, 2, 3, and 4 and the corresponding P chain framework regions 1, 2, 3, and 4 of any of the anti-NPMlc TCRs set forth in Table 1. As an example, a TCR can have the a chain framework regions 1, 2, 3, and 4 of SEQ ID NO:32 and the P chain framework regions 1, 2, 3, and 4 of SEQ ID NO:40 or the a chain framework regions 1, 2, 3, and 4 of SEQ ID NO:68 and the P chain framework regions 1, 2, 3, and 4 of SEQ ID NOL76.

[0091] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include an a chain that includes an amino acid sequence having at least 90 percent identity to an amino acid sequence of an a chain variable (V) region of any of the anti-NPMlc TCRs set forth in Table 1, and a P chain that includes an amino acid sequence having at least 90 percent identity to an amino acid sequence of a corresponding p chain variable (V) region as set forth in Table 1. A anti-NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to an amino acid sequence of an a chain variable (V) region of any of the anti-NPMlc TCRs set forth in Table 1, and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to an amino acid sequence of a corresponding P chain variable (V) region as set forth in Table 1. In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include (a) an a chain that includes an amino acid sequence having 100 percent identity to an amino acid sequence of an a chain variable (V) region of any of the anti-NPMlc TCRs set forth in Table 1, and (b) a P chain that includes an amino acid sequence having 100 percent identity to an amino acid sequence of a corresponding p chain variable (V) region as set forth in Table 1. As an example, an anti- NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 percent identity to the amino acid sequence SEQ ID NO:32, and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 percent identity to an amino acid sequence SEQ ID NO:40. As another example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 percent identity to the amino acid sequence SEQ ID NO: 68, and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 percent identity to an amino acidsequence SEQ ID NO: 76.

[0092] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include (a) an a chain that includes an amino acid sequence having at least 90 percent identity to an amino acid sequence of an a chain variable (V) region of any of the anti-NPMlc TCRs set forth in Table 1, provided that the a chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding a chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1, and (b) a P chain that includes an amino acid sequence having at least 90 percent identity to an amino acid sequence of a corresponding P chain variable (V) region as set forth in Table 1, provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding P chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1. An anti-NPMlc TCR or antigen binding fragment thereof provided herein can include (a) an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to an amino acid sequence of an a chain variable (V) region of any of the anti-NPMlc TCRs set forth in Table 1, provided that the a chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding a chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1, and (b) a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to an amino acid sequence of a corresponding P chain variable (V) region as set forth in Table 1, provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding P chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1. As an example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence of SEQ ID NO:32, provided that the a chain includes CDR-1, CDR-2, and CDR- 3 amino acid sequences as set forth in SEQ ID NOs:29, 30, and 31, respectively, and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to an amino acid sequence of SEQ ID NO:40, provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences as set forth in SEQ ID NOs:37, 38, and 39, respectively. As another example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can include an a chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence of SEQ ID NO: 68, provided that the a chain includes CDR-1, CDR-2, and CDR-3 amino acid sequencesas set forth in SEQ ID NOs:65, 66, and 67, respectively, and a P chain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to an amino acid sequence of SEQ ID NO:76, provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences as set forth in SEQ ID NOs:73, 74, and 75, respectively

[0093] In some embodiments, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule can include (a) an a chain having an amino acid sequence of an a chain variable (V) region of any of the anti-NPMlc TCRs set forth in Table 1 or an amino acid of an a chain variable (V) region of any of the anti- NPMlc TCRs set forth in Table 1 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions) and (b) a P chain having the amino acid sequence of a corresponding p chain variable (V) region as set forth in Table 1 or an amino acid of a corresponding p chain variable (V) region as set forth in Table 1 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). A anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) in the context of an MHC molecule, can include (a) an a chain having the amino acid sequence of any of the TCRs set forth in Table 1 with zero, one, two, three, four, five, six, seven, eight, nine, or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the a chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding a chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1, and (b) a P chain having the amino acid sequence of a corresponding p chain variable (V) region as set forth in Table 1, with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences that are 100% identical to the corresponding P chain CDR-1, CDR-2, and CDR-3 sequences set forth in Table 1. As an example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule, can include an a chain having the amino acid sequence set forth in SEQ ID NO:32 with zero, one, two, three, four, five, six, seven, eight, nine, or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the a chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences as set forth in SEQ ID NOs:29,30, and 31, respectively, and a P chain having the amino acid sequence set forth in SEQ ID NO:40, respectively, with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences as set forth in SEQ ID NOs:37, 38, and 39, respectively. As another example, an anti-NPMlc TCR or antigen binding fragment thereof provided herein having the ability to specifically bind CLAVEEVSL (SEQ ID NO:5) in the context of an MHC molecule, can include an a chain having the amino acid sequence set forth in SEQ ID NO: 68 with zero, one, two, three, four, five, six, seven, eight, nine, or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the a chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences as set forth in SEQ ID NOs:65, 66, and 67, respectively, and a P chain having the amino acid sequence set forth in SEQ ID NO:76, respectively, with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that the P chain includes CDR-1, CDR-2, and CDR-3 amino acid sequences as set forth in SEQ ID NOs:73, 74, and 75, respectively

[0094] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein contains a Va or Vy region that contains:(a) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: l l, 12, and 13, respectively;(b) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:29, 30, and 31, respectively;(c) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:47, 48, and 49, respectively;(d) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:65, 66, and 67, respectively;(e) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:83, 84, and 85 respectively;(f) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 101, 102, and 103, respectively;(g) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 119, 120, and 121, respectively;(h) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 137, 138, and 139, respectively;(i) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 155, 156, and 157, respectively;(j) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 173, 174, and 175, respectively;(k) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:191, 192, and 193, respectively;(l) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:209, 210, and 211, respectively;(m) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:227, 228, and 229, respectively;(n) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:245, 246, and 247, respectively;(o) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:263, 264, and 265, respectively; or(p) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:281, 282, and 283, respectively.Also, among the provided anti-NPMlc TCRs are those having sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences.

[0095] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein contains a VP or V5 region that contains:(a) a CDR-1, CDR-2, and CDR-3 comprising the SEQ ID NOs: 19, 20, and 21, respectively;(b) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:37, 38, and 39, respectively;(c) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:55, 56, and 57, respectively;(d) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:73, 74, and 75, respectively;(e) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:91, 92, and 93, respectively;(f) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 109, 110, and 111, respectively;(g) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 127, 128, and 129, respectively;(h) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 145, 146, and 147, respectively;(i) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 163, 164, and 165, respectively;(j) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 181, 182, and 183, respectively;(k) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs: 199, 200, and 201, respectively;(l) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:217, 218, and 219, respectively;(m) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:235, 236, and 237, respectively;(n) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:253, 254, and 255, respectively;(o) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:271, 272, and 273, respectively;(p) a CDR-1, a CDR-2, and a CDR-3, comprising SEQ ID NOs:289, 290, and 291, respectively.Also, among the provided anti-NPMlc TCRs are those having sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences.

[0096] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein includes a Va or Vy region that contains a CDR-1, a CDR-2, and a CDR-3, comprising a CDR-1, a CDR-2, and a CDR-3 amino acid sequence, respectively, set forth in Table 1, such as in each row therein and a VP or V5 region that contains a CDR-1, a CDR-2, and a CDR-3, comprising a CDR-1, a CDR-2, and a CDR-3 amino acid sequence, respectively, set forth in Table 1, such as in each row therein.

[0097] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein includes a Va or Vy region that contains a CDR-1, a CDR-2, and a CDR-3, comprising a CDR-1, a CDR-2, and a CDR-3 amino acid sequence, respectively, contained within a Va or Vy region amino acid sequence set forth in Table 1, such as in each row therein, and a VP or V5 region that contains a CDR-1, a CDR-2, and a CDR-3, comprising a CDR-1, a CDR-2, and a CDR-3 amino acid sequence, respectively, contained within a VP or V5 region amino acid sequence set forth in Table 1, such as in each row therein. In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof provided herein includes a Va or Vyregion amino acid sequence and a corresponding V0 or V5 region amino acid sequence as set forth in Table 1, such as in each row therein. Also, among the provided anti-NPMlc TCRs are those containing sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. Exemplary anti-NPMlc TCRs containing such CDRs, or their modified versions as described elsewhere herein, also are set forth in the Table 1, such as in each row therein.Table 1. SEQ ID NOs for Amino Acid Sequences of CDRs and Variable Regions of anti-NPMlc TCRs (sequence are provided in Table 12).

[0098] In some examples, an anti-NPMlc TCR or antigen binding fragment thereof provided herein can be designed to include an a chain (or y chain) that includes a set of three CDRs (e.g., a CDR-1, CDR-2, and CDR-3) as set forth in Table 1 (e.g, SEQ ID NOs:29-31, SEQ ID NOs: 65-67, SEQ ID NOs: 83 -85, or SEQ ID NOs: 101-103) and a 0 chain (or 5 chain) that includes a set of three CDRs (e.g., a CDR-1, CDR-2, and CDR-3) as set forth in Table 1 (e.g, SEQ ID NOs:37-39, SEQ ID NOs:73-75, SED ID NOs:92-92, or SEQ ID NOs: 109-l l l).

[0099] In some embodiments, the Va region comprises a CDR-1 comprising SEQ ID NO:29,a CDR-2 comprising SEQ ID NO:30, and a CDR-3 comprising SEQ ID NO:31, and the VP region comprises a CDR-1 comprising SEQ ID NO:37, a CDR-2 comprising SEQ ID NO:38, and a CDR-3 comprising SEQ ID NO:39. In some embodiments, the Va region comprises a CDR-1 comprising SEQ ID NO:65, a CDR-2 comprising SEQ ID NO:66, and a CDR-3 comprising SEQ ID NO:67, and the VP region comprises a CDR-1 comprising SEQ ID NO:73, a CDR-2 comprising SEQ ID NO:74, and a CDR-3 comprising SEQ ID NO:75. In some embodiments, the Va region comprises a CDR-1 comprising SEQ ID NO:83, a CDR-2 comprising SEQ ID NO:84, and a CDR-3 comprising SEQ ID NO:85, and the VP region comprises a CDR-1 comprising SEQ ID NO:92, a CDR-2 comprising SEQ ID NO:93, and a CDR-3 comprising SEQ ID NO:94. In some embodiments, the Va region comprises a CDR-1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and the VP region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR- 2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111.

[0100] In some embodiments, the Va region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va region sequence of SEQ ID NO:32, and the VP region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP region sequence of SEQ ID NO:40. In some embodiments, the Va region comprises a CDR-1, a CDR- 2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va region sequence of SEQ ID NO:68, and the VP region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP region sequence of SEQ ID NO:76. In some embodiments, the Va region comprises a CDR- 1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va region sequence of SEQ ID NO:86, and the VP region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP region sequence of SEQ ID NO:94. In some embodiments, the Va region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va region sequence of SEQ ID NO: 104, and the VP region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP region sequence of SEQ ID NO:112.

[0101] In some embodiments, the Va region comprises SEQ ID NO:32 or a sequence that has at least 90% sequence identity thereto, and the VP region comprises SEQ ID NO:40 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the Va region comprises SEQ ID NO:68 or a sequence that has at least 90% sequence identity thereto, andthe VP region comprises SEQ ID NO:76 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the Va region comprises SEQ ID NO:86 or a sequence that has at least 90% sequence identity thereto, and the VP region comprises SEQ ID NO:94 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the Va region comprises SEQ ID NO: 104 or a sequence that has at least 90% sequence identity thereto, and the VP region comprises SEQ ID NO: 112 or a sequence that has at least 90% sequence identity thereto.

[0102] In some embodiments, the Va region comprises SEQ ID NO:32, and the VP region comprises SEQ ID NO:40. In some embodiments, the Va region comprises SEQ ID NO:68, and the VP region comprises SEQ ID NO:76. In some embodiments, the Va region comprises SEQ ID NO:86, and the VP region comprises SEQ ID NO:94. In some embodiments, the Va region comprises SEQ ID NO: 104, and the VP region comprises SEQ ID NO: 112.C. Exemplary Constant Domains

[0103] In some embodiments, the a chain of an anti-NPMlc TCR or antigen-binding fragment thereof provided herein further contains an a constant (Ca) region or portion thereof. In some aspects, the P chain further contains a P constant (CP) region or portion thereof. Thus, in some embodiments, an anti-NPMlc TCR provided herein (e.g., an anti-AIQDLCLAV or anti-CLAVEEVSL TCR provided herein) or an antigen-binding fragment thereof contains an a chain comprising a Va region and a Ca domain or portion thereof and / or a P chain comprising a VP region and a CP domain or portion thereof. In some embodiments, the y chain of an anti- NPMlc TCR or antigen-binding fragment thereof provided herein further contains a y constant (Cy) region or portion thereof. In some aspects, the 5 chain further contains a 5 constant (C5) region or portion thereof. Thus, in some embodiments, an anti-NPMlc TCR provided herein (e.g., an anti-AIQDLCLAV or anti-CLAVEEVSL TCR provided herein) or an antigen-binding fragment thereof contains a y chain comprising a Vy region and a Cy domain or portion thereof and / or a 5 chain comprising a V5 region and a C5 domain or portion thereof.

[0104] In some embodiments, the a chain and the P chain, or the y chain and the 5 chain, of an anti-NPMlc TCR provided herein each further contain a constant domain. In some embodiments, the Ca domain and CP domain, or the Cy domain and C5 domain, individually are mammalian (e.g., a human or murine constant domain). In some embodiments, the constant domain is adjacent to the cell membrane. For example, in some cases, the extracellular portion of the anti-NPMlc TCR formed by the two chains contains two membrane-proximal constant domains, and two membrane-distal variable domains, which variable domains each containCDRs.

[0105] In some aspects, provided herein are anti-NPMlc TCRs that contain a human constant domain, such as an a chain containing a human Ca domain and a P chain containing a human CP domain, or a y chain containing a human Cy domain and a 5 chain containing a human C5 domain. In some embodiments, the provided anti-NPMlc TCRs are fully human. Among the provided anti-NPMlc TCRs are TCRs containing a human constant domain, such as fully human TCRs, whose expression and / or activity, such as when expressed in human cells, e.g., human T cells, such as primary human T cells, are not impacted by or are not substantially impacted by the presence of an endogenous human TCR.

[0106] In some embodiments, each of the Ca and the CP domains, or each of the Cy and the C5 domains, is human. In some embodiments, the Ca is encoded by the TRAC gene (IMGT nomenclature) or is a variant thereof. In some embodiments, the CP is encoded by TRBC1 or TRBC2 genes (IMGT nomenclature) or is a variant thereof. In some embodiments, the Cy is encoded by the TRGC1 or TRGC2 genes (IMGT nomenclature) or is a variant thereof. In some embodiments, the C5 is encoded by TRDC genes (IMGT nomenclature) or is a variant thereof.

[0107] In some embodiments, the Ca domain or a variant thereof has or comprises the sequence of amino acids set forth in SEQ ID NO: 3, or a sequence of amino acids that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:3. In some embodiments, the Ca domain has or comprises the sequence of amino acids set forth in SEQ ID NO:3. In some embodiments, the CP domain or variant thereof has or comprises the sequence of amino acids set forth in SEQ ID NO:7, or a sequence of amino acids that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:7. In some embodiments, the CP domain has or comprises the sequence of amino acids set forth in SEQ ID NO:7. In some embodiments, the anti-NPMlc TCR comprises a Ca domain and a CP domain set forth in SEQ ID NO:3 and 7, respectively.

[0108] In some embodiments, the Cy domain or a variant thereof has or comprises the sequence of amino acids set forth in SEQ ID NO: 312 or 313, or a sequence of amino acids that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:312 or 312. In some embodiments, the Cy domain has or comprises the sequence of amino acids set forth in SEQ ID NO:312. In some embodiments, the Cy domain has or comprises the sequence of amino acids set forth in SEQ ID NO:313. In some embodiments, the C5 domain or variant thereof has or comprises the sequence of amino acids set forth in SEQ ID NO: 314 or a sequence of amino acids that exhibitsat least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:314. In some embodiments, the anti-NPMlc TCR comprises a Cy domain and a C5 domain set forth in SEQ ID NO:312 and 314, respectively. In some embodiments, the anti-NPMlc TCR comprises a Cy domain and a C5 domain set forth in SEQ ID NO:313 and 314, respectively.

[0109] In some embodiments, the variant of a Ca domain contains replacement of at least one non-native cysteine, such as any replacement described herein. In some embodiments, the variant of a CP domain contains replacement of at least one non-native cysteine, such as any replacement described herein.

[0110] In some embodiments, any of the provided anti-NPMlc TCRs or antigen-binding fragments thereof can be a human / mouse chimeric TCR. In some embodiments, an anti- NPMlc TCR or antigen-binding fragment thereof provided herein comprises an a chain and / or a P chain, or a y chain and / or a 5 chain, comprising a mouse constant domain. In some embodiments, the Ca domain and / or the CP domain, or the Cy domain and / or the C5 domain, are a mouse Ca domain and / or a mouse CP domain, or a mouse Cy domain and / or a mouse C5 domain. In some embodiments, the Ca domain and / or the CP domain, or the Cy domain and / or the C5 domain, is or comprises any Ca domain and / or CP domain, or Cy domain and / or C5 domain described in WO2015 / 184228, WO2015 / 009604, or WO2015 / 009606.[OHl] In some embodiments, an anti-NPMlc TCR or antigen-binding fragment thereof comprises a variant of an a chain and / or a P chain, or a y chain and / or a 5 chain. In some embodiments, the variant comprises the amino acid sequence of any of the anti-NPMlc TCRs described herein with one, two, three, or four or more amino acid substitution(s) in the constant domain of the a or P chain. In some embodiments, the anti-NPMlc TCRs (or functional portions thereof) comprising the substituted amino acid sequence(s) advantageously provide one or more of decreased mispairing with an endogenous TCR chain(s), increased expression by a host cell, increased recognition of AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5), and increased anti-tumor activity as compared to the parent TCR comprising an unsubstituted amino acid sequence.

[0112] In some embodiments, the anti-NPMlc TCR may be a heterodimer of the a and P chains, or the y and 5 chains that are linked, such as by a disulfide bond or disulfide bonds. In some embodiments, the constant domain of the anti-NPMlc TCR may contain short connecting sequences in which a cysteine residue forms a disulfide bond, thereby linking the two chains of the TCR. In some embodiments, an anti-NPMlc TCR may have an additional cysteine residue in each of the a and P chains, or the y and 5 chains, such that the TCR containstwo disulfide bonds in the constant domains. In some embodiments, each of the constant and variable domains contains disulfide bonds formed by cysteine residues.

[0113] In some embodiments, an anti-NPMlc TCR provided herein can contain an introduced disulfide bond or bonds. In some embodiments, the native disulfide bonds are not present. In some embodiments, the one or more of the native cysteines (e.g., in the constant domain of the a chain and the P chain, or the y chain and the 5 chain) that form a native interchain disulfide bond are substituted to another residue, such as to a serine or alanine. In some embodiments, an introduced disulfide bond can be formed by mutating non-cysteine residues on the a and P chains, such as in the constant domain of the a chain and the P chain, or the y chain and the 5 chain, to cysteine. Opposing cysteines in the TCR a and P chains, or TCR y and 5 chains provide a disulfide bond that links the constant domains of TCR a and P chains, or TCR y and 5 chains of the substituted TCR to one another and which is not present in a TCR comprising the unsubstituted constant domain in which the native disulfide bonds are present, such as unsubstituted native human constant domain or the unsubstituted native mouse constant domain. In some embodiments, the presence of non-native cysteine residues (e.g., resulting in one or more non-native disulfide bonds) in a recombinant TCR can favor production of the desired recombinant TCR in a cell in which it is introduced over expression of a mismatched TCR pair containing a native TCR chain. In some embodiments, the a or y chain and the P or 5 chain of an anti-NPMlc TCR each contains a cysteine amino acid substitution. The cysteine substitution can facilitate the formation of a disulfide bond between the a and P chains or between the y and 5 chains. The formation of the disulfide bond can facilitate pairing between the a and P chains or between the y and 5 chains, and / or decrease mispairing with an endogenous TCR chain(s). Suitable cysteine substitutions include, but are not limited to, a T47C substitution in the a constant domain (position relative to SEQ ID NO: 3) chain and a S56C substitution (positions relative to SEQ ID NO:7 or 9) in the P chain.

[0114] Exemplary non-native disulfide bonds of a TCR are described in published International PCT Patent Application Nos. W02006 / 000830 and W02006 / 037960. In some embodiments, cysteines can be introduced or substituted at a residue corresponding to Thr48 of the Ca domain (corresponding to Thr47 of SEQ ID NO:3) and Ser57 of the CP domain (corresponding to Ser56 of SEQ ID NO:7 or 9), at residue Thr45 of the Ca domain and Ser77 of the CP domain, at residue TyrlO of the Ca domain and Seri 7 of the CP domain, at residue Thr45 of the Ca domain and Asp59 of the CP domain and / or at residue Seri 5 of the Ca domain and Glut 5 of the CP domain.

[0115] In some embodiments, any of the provided cysteine mutations can be made at acorresponding position in another sequence, for example, in a human or mouse Ca domain and / or CP domain, or Cy domain and / or C5 domain, sequence described above. The term “corresponding” with reference to positions of a protein, such as recitation that amino acid positions “correspond to” amino acid positions in a disclosed sequence, such as set forth in the Sequence Listing, refers to amino acid positions identified upon alignment with the disclosed sequence based on structural sequence alignment or using a standard alignment algorithm, such as the GAP algorithm. For example, corresponding residues can be determined by alignment of a reference sequence with the Ca sequence set forth in SEQ ID NO:3, or the CP sequence set forth in SEQ ID NO: 7 or 9, by structural alignment methods as described herein. By aligning the sequences, one can identify corresponding residues, for example, using conserved and identical amino acid residues as guides.

[0116] In some embodiments, an anti-AIQDLCLAV TCR or antigen-binding fragment thereof provided herein comprises an a or y chain that is or comprises the an amino acid sequence as set forth in any of SEQ ID NOs: 17, 35, 53, or a sequence that has at least 90% sequence identity thereto, such as a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence and / or a corresponding (see Table 2) P or 5 chain that is or comprises an amino acid sequence as set forth in any of SEQ ID NO:25, 43, and 63, or a sequence that has at least 90% sequence identity thereto, such as a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence.

[0117] In some embodiments, an anti-CLAVEEVSL TCR or antigen-binding fragment thereof provided herein comprises an a or y chain that is or comprises the an amino acid sequence as set forth in any of SEQ ID NOs:71, 89, 107, 125, 143, 161, 179, 197, 215, 233, 251, 269, or 287, or a sequence that has at least 90% sequence identity thereto, such as a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence and / or a corresponding (see Table 2) P or 5 chain that is or comprises an amino acid sequence as set forth in any of SEQ ID NO:79, 97, 115, 133, 151, 169, 187, 205, 223, 241, 259, 277, or 295, or a sequence that has at least 90% sequence identity thereto, such as a sequence having at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with such a sequence.

[0118] Exemplary anti-NPMlc TCRs or antigen-binding fragments include those set forth in Table 2, such as in each row therein. In some embodiments, the Va and VP region, or the Vy and V5 region, contain the amino acid sequences corresponding to the SEQ ID NOs: set forth in Table 2, such as in each row therein. In some embodiments, the Va and VP region, or theVy and V5 region, contain the CDR-1, the CDR-2 and the CDR-3 sequences contained within the Va and VP region, set forth in Table 2, such as in each row therein. In some aspects, the anti-NPMlc TCR contains constant a and constant P domain sequences, such as those corresponding to the SEQ ID NOs: set forth in Table 2, such as in each row therein. In some embodiments, the anti-NPMlc TCR contains a full sequence comprising the variable and constant domains, such as a sequence corresponding to the SEQ ID NOs: set forth in Table 2 (“Full a-P2A-P”), such as in each row therein. Also, among the provided anti-NPMlc TCRs are those containing sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. Exemplary anti-NPMlc TCRs containing such sequences, or their modified versions as described elsewhere herein, also are set forth in the Table 2, respectively, such as in each row therein. In some aspects, the provided exemplary anti-NPMlc TCRs, when expressed as a mature protein, comprises the mature Va and / or mature VP region, or the mature Vy and / or V5 mature region, for example, without the signal sequence (e.g., from cleavage of the signal sequence) when fully processed and expressed.Table 2: SEQ ID NOs of Amino Acid Sequences of Variable and Constant Regions of anti- NPMlc TCRs.

[0119] In some embodiments, the a or y chain comprises SEQ ID NO:35, or a sequence thathas at least 90% sequence identity thereto, and the P or 5 chain comprises SEQ ID NO:43, or a sequence that has at least 90% sequence identity thereto. In some embodiments, the a or y chain comprises SEQ ID NO:71, or a sequence that has at least 90% sequence identity thereto, and the P or 5 chain comprises SEQ ID NO:79, or a sequence that has at least 90% sequence identity thereto. In some embodiments, the a or y chain comprises SEQ ID NO:89, or a sequence that has at least 90% sequence identity thereto, and the P or 5 chain comprises SEQ ID NO: 97, or a sequence that has at least 90% sequence identity thereto. In some embodiments, the a or y chain comprises SEQ ID NO: 107, or a sequence that has at least 90% sequence identity thereto, and the P or 5 chain comprises SEQ ID NO: 115, or a sequence that has at least 90% sequence identity thereto.

[0120] In some embodiments, the a chain comprises SEQ ID NO:35, and the P chain comprises SEQ ID NO:43. In some embodiments, the a chain comprises SEQ ID NO:71 and the P chain comprises SEQ ID NO:79. In some embodiments, the a chain comprises SEQ ID NO:89 and the P chain comprises SEQ ID NO:97. In some embodiments, the a chain comprises SEQ ID NO: 107 and the P chain comprises SEQ ID NO: 115.

[0121] In some embodiments, the anti-AIQDLCLAV TCR comprises the amino acid sequence of SEQ ID NO:27, 45, or 63, or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:27, 45, or 63. In some embodiments, the anti-CLAVEEVSL TCR comprises the amino acid sequence of SEQ ID NO: 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, or 297, or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, or 297.

[0122] In some embodiments, an anti-NPMlc TCR or antigen binding portion thereof provided herein may be a recombinantly produced natural protein or a mutated form thereof in which one or more properties, such as a binding characteristic, has been altered. In some aspects, the nucleic acid is synthetic.

[0123] In some embodiments, any of the described anti-NPMlc TCRs (or nucleic acids encoding the anti-NPMlc TCRs) can be modified to increase affinity of the TCR for the target neoantigen, to improve stability of the TCR, and / or to modify a function of the TCR, such as signaling. Methods for modifying the anti-NPMlc TCR include, but are not limited to, alanine scanning and peptide sequence optimization. In some embodiments, any of the described anti- NPMlc TCRs can be modified to contain a disulfide bond in the constant a TCR region.D. anti-NPMl TCR single chain variable fragments

[0124] Any of the anti-NPMl TCR CDRs or variable regions described herein can be usedin the formation of single chain variable fragments (scFvs), bispecific T cell engagers (BiTEs), chimeric T cell receptors (CARs), or other proteins containing scFvs, BiTEs or CARs.

[0125] An “scFv” is a fusion protein comprising a variable heavy chain region (VH) and a variable light chain region (VL) of an immunoglobulin or a variable a region (Va) and a variable P region (VP) of a TCR connected with a short linker peptide. An scFv retains the antigen binding properties of the intact immunoglobulin or TCR from with the variable regions are derived. Nucleic acids encoding the anti-AIQDLCLAV or CLAVEEVSL scFvs are also contemplated.

[0126] In some embodiments, anti-AIQDLCLAV and anti -CLAVEEVSL scFvs are described comprising a Va region and a VP region of any of the described anti-AIQDLCLAV or anti-CLAVEEVSL TCRs, wherein the Va region and the VP region are linked via a linker peptide and wherein the anti-AIQDLCLAV or anti-CLAVEEVSL scFv retains the antigen binding properties of the anti-AIQDLCLAV or anti-CLAVEEVSL TCR from which the variable regions are derived. Nucleic acids encoding the anti-AIQDLCLAV and anti- CLAVEEVSL scFvs are also contemplated. An anti-AIQDLCLAV or anti-CLAVEEVSL scFv can be formed by linking the C terminus of the Va chain with the N terminus of the VP chain. Alternatively, the C terminus of the VP can be linked to the N-terminus of the Va chain. The peptide linker can be about 10 to about 25 amino acids. In some embodiments, the scFv peptide linker is rich in glycine. An scFv peptide linker can be, but is not limited to, (G4S)Xwhere x is an integer from 2 to 5 (inclusive). In some embodiments, the scFv peptide linked comprises Gly-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser (i.e., also termed [(Gly)4Ser]3, (G4S)s or G4S (x3)). In some embodiments, the scFv peptide linker consists of G4S (X3).

[0127] Bispecific T cell engagers (BiTEs) are recombinant molecules comprising two flexibly linked antigen binding domains, such as scFvs. In a typical BiTE, one antigen binding domain of BiTE is specific for CD3 or other activating antigen on an immune cell, and the second antigen binding domain has affinity for a second antigen, such as a tumor antigen or antigen on a target cell. BiTEs can be used to target T cells, which contain a CD3 receptor with a target cell (as described in WO99054440, W02005040220, and W02008119567). BiTEs are capable of binding T cells transiently to target cells and, at the same time, activating the cytolytic activity of the T cells.

[0128] In some embodiments, anti-AIQDLCLAV and anti-CLAVEEVSL BiTE molecules are described comprising a first antigen binding domain (e.g., scFv) specific for an activating antigen on an immune cell and a second antigen binding domain comprising an AIQDLCLAVor CLAVEEVSL binding domain or scFv comprising the variable regions or CDRs of any of the described anti-AIQDLCLAV or anti -CLAVEEVSL TCRs. In some embodiments, a BiTE comprises an anti-CD3 scFv and an anti-AIQDLCLAV or anti-CLAVEEVSL scFv comprising the Va region and a VP region of any of the described anti-AIQDLCLAV or anti- CLAVEEVSL TCRs. Nucleic acids encoding the anti-AIQDLCLAV or anti-CLAVEEVSL BiTEs are also contemplated.

[0129] A “chimeric antigen receptor” or “CAR” is a recombinant protein comprising an antigen-binding domain (e.g., an antigen-binding fragment of any of the described anti-NPMlc TCRs) linked to a cell signaling and / or cell activation domain via a transmembrane domain. The cell -signaling domain can be, but is not limited to, a T-cell signaling domain. When utilized in a CAR, the antigen-binding fragment of a described anti-NPMlc TCR can be provided as an scFv. The CAR can be a first generation CAR T cell, a second generation CAR T cell, a third generation CAR T cell, a fourth generation CAR T cell, dual-antigen receptor CAR T cell, or a CAR T cell having an inducible suicide gene, or a combination thereof. The CAR can have a single signaling and / or cell activation domain, multiple signaling and / or cell activation domains, or one or more signaling and / or cell activation domains and one or more co-stimulation domains. The signaling and / or cell activation domain or co-stimulation domains can be, but are not limited to, CD3zeta domain, CD28 domain, a CD 137 (4-1BB) domain, an ICOS domain, a CD27 domain, an 0X40 domain, a LFA1 domain, a PD-1 domain, a CD 150 domain, a CD244 domain, a NKG2D domain, and A DAP 10 domain. In some embodiments, a CAR has a CD28 transmembrane domain.

[0130] In some embodiments, anti-AIQDLCLAV and anti-CLAVEEVSL CARs are described comprising an antigen binding fragment of any of the described anti-AIQDLCLAV or anti-CLAVEEVSL TCRs or an scFv comprising the variable regions or CDRs of any of the described TCRs, a transmembrane domain, and a signaling and / or cell activation domain. In some embodiments, the antigen binding fragment of any of the described anti-AIQDLCLAV or anti-CLAVEEVSL TCRs comprises an anti-AIQDLCLAV or anti-CLAVEEVSL scFv, wherein the anti-AIQDLCLAV or anti-CLAVEEVSL scFv comprises a Va region and a VP region of any of the described anti-AIQDLCLAV or anti-CLAVEEVSL TCRs. The anti- AIQDLCLAV or anti-CLAVEEVSL CAR can be, but is not limited to, a first generation CAR T cell, a second generation CAR T cell, a third generation CAR T cell, a fourth generation CAR T cell, dual-antigen receptor CAR T cell, or a CAR T cell having an inducible suicide gene, or a combination thereof. Nucleic acids encoding the anti-AIQDLCLAV or anti- CLAVEEVSL CARs are also contemplated.II. NUCLEIC ACIDS ENCODING anti-NPMlc TCRs

[0131] Also provided herein are nucleic acids, such as polynucleotides or nucleic acid molecules, encoding any of the described anti-NPMlc TCRs or antigen-binding fragments thereof. The nucleic acids may include those encompassing natural and / or non-naturally occurring nucleotides and bases, e.g., including those with backbone modifications. The terms “nucleic acid molecule,” “nucleic acid,” and “polynucleotide” may be used interchangeably, and refer to a polymer of nucleotides. Such polymers of nucleotides may contain natural and / or non-natural nucleotides, and include, but are not limited to, DNA and RNA. “Nucleic acid sequence” refers to the linear sequence of nucleotides that comprise the nucleic acid molecule or polynucleotide.

[0132] In some embodiments, the nucleic acid is or contains cDNA. In some aspects, the polynucleotides can be modified for use in a construct described herein, such as for codon optimization. In some embodiments, the nucleic acid sequences can be designed to contain terminal restriction site sequences for purposes of cloning into vectors.

[0133] In some embodiments, an anti-NPMlc TCR or antigen-binding portion thereof provided herein can be synthetically generated from knowledge of the amino acid sequence of the TCR.

[0134] Described are nucleic acid sequences that encode an Va region, a VP region, a Va region and a VP region, an a chain a P chain or an a chain and a P chain. In some embodiments, the nucleic acid sequences can encode a Vy region, a V5 region, a Vy region and a V5 region, a y chain, a 5 chain, or a y chain and a 5 chain.

[0135] The nucleotide sequences can be provided as a pair of nucleotide sequences comprising a first nucleotide sequence encoding a Va region or an a chain and a second nucleotide sequence encoding a VP region or a P chain; or a first nucleotide sequence encoding a Vy region or a y chain and a second nucleotide sequence encoding a V5 region or a 5 chain. The first and second nucleotide sequences can be provided on separate expression cassettes, plasmids, or vectors (e.g., viral vectors) or on a single expression cassette, plasmid, or vector.

[0136] In some embodiments, a polynucleotide contains nucleic acid sequences encoding an Va region or a chain and a VP region or P chain (or a Vy region or y chain and a V5 region or 5 chain) wherein the nucleic acid sequence encoding the Va (or Vy) region or a (or y) chain is linked to the sequence encoding the VP (or V5) region or P (or 5) chain by a sequence encoding a peptide sequence that causes ribosome skipping or self-cleavage (e.g., a 2 A peptide, e.g., a P2A peptide, a T2A peptide, a F2A peptide, or an E2A peptide) or an (internal ribosome entry site (IRES) sequence. In some embodiments, the sequences encoding the Va and VP regionsor a and P chains (or Vy and V5 regions or y and 5 chains) are operably linked to a single promoter. In some embodiments, the sequences encoding the Va and VP regions or a and P chains (or Vy and V5 regions or y and 5 chains) are each operably linked to a single bidirectional promoter (e.g., a PGK promoter). In some embodiments, the sequence encoding the Va (or Vy) region or the a (or y) chain can be 5' of the sequence encoding the VP (or V5) region or the P (or 5) chain. In some embodiments, the sequence encoding the VP (or V5) region or the P (or 5) chain can be 5' of the sequence encoding the Va (or Vy) region or the a (or y) chain.

[0137] In some embodiments, the nucleotide sequence encoding the a or y chain and / or the nucleotide sequence encoding the P or 5 chain, or any domains, regions (e.g., Va and VP regions) or portions thereof, is codon-optimized. Typically, codon optimization involves balancing the percentages of codons selected with the published abundance of human transfer RNAs so that none is overloaded or limiting. Most amino acids are encoded by more than one codon, and codon usage varies from organism to organism. Differences in codon usage between transfected genes and host cells can have effects on protein expression and immunogenicity of a nucleic acid construct. In general, for codon optimization, codons are chosen to select for those codons that are in balance with human usage frequency. Typically, the redundancy of the codons for amino acids is such that different codons code for one amino acid. In some embodiments, in selecting a codon for replacement, it may be desired that the resulting mutation is a silent mutation such that the codon change does not affect the amino acid sequence.

[0138] In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO:33 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO:41 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO:69 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO:77 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO:87 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO:95 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO: 105 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO: 113 or a sequence that has at least 90% sequence identity thereto.

[0139] In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO:34 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO:42 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO:70 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO:78 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO:88 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO:96 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the Va region comprises SEQ ID NO: 106 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the VP region comprises SEQ ID NO: 114 or a sequence that has at least 90% sequence identity thereto.

[0140] In some embodiments, the nucleotide sequence encoding the a or y chain comprises SEQ ID NO:36 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the P or 5 chain comprises SEQ ID NO:44 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the a or y chain comprises SEQ ID NO:72 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the P or 5 chain comprises SEQ ID NO:80 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the a or y chain comprises SEQ ID NOVO or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the P or 5 chain comprises SEQ ID NO:98 or a sequence that has at least 90% sequence identity thereto. In some embodiments, the nucleotide sequence encoding the a or y chain comprises SEQ ID NO: 108 or a sequence that has at least 90% sequence identity thereto, and the nucleotide sequence encoding the P or 5 chain comprises SEQ ID NO: 116 or a sequence that has at least 90% sequence identity thereto. Also, among the provided nucleic acid(s) or polynucleotides provided herein are those containing sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences.

[0141] In some embodiments, the nucleotide sequence encoding the a chain comprises SEQ ID NO:36, and the nucleotide sequence encoding the P chain comprises SEQ ID NO:44. In some embodiments, the nucleotide sequence encoding the a chain comprises SEQ ID NO: 72 and the nucleotide sequence encoding the P chain comprises SEQ ID NO:80. In some embodiments, the nucleotide sequence encoding the a chain comprises SEQ ID NOVO and thenucleotide sequence encoding the P chain comprises SEQ ID NO:98. In some embodiments, the nucleotide sequence encoding the a chain comprises SEQ ID NO: 108 and the nucleotide sequence encoding the P chain comprises SEQ ID NO: 116.

[0142] In some embodiments, the nucleic acid sequence encoding the a chain comprises any one of SEQ ID NO: 15, 33, or 51, or a degenerate sequence thereof, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto and the nucleotide sequence encoding the P chain comprises any one of corresponding SEQ ID NOs:23, 41, and 59, or a degenerate sequence thereof or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity thereto.

[0143] In some embodiments, the nucleic acid sequence encoding the a chain comprises any one of SEQ ID NO: 69, 87, 105, 123, 141, 159, 177, 195, 213, 231, 249, 267, or 285, or a degenerate sequence thereof, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto the nucleotide sequence encoding the P chain comprises any one of corresponding SEQ ID NOs: 77, 95, 113, 131, 149, 167, 185, 203, 221, 239, 257, 275, or 293, or a degenerate sequence thereof, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto.

[0144] In some embodiments, the a or y chain and / or P or 5 chain of the anti-NPMlc TCR is encoded by a sequence of nucleotides comprising a signal peptide (also called a leader sequence). Non-limiting examples of such a signal peptide are signal peptides that have or comprise the sequence of amino acids set forth in any of SEQ ID NOs:315-624.

[0145] In some embodiments, the nucleic acid encoding the a or y chain and the nucleic acid encoding the P or 5 chain can be connected via a linker, such as any described elsewhere herein.

[0146] In some embodiments, the nucleic acid encoding the Va or Vy region or a or y chain and the nucleic acid encoding the VP or V5 region or p or 5 chain can be connected via a cleavable linker sequence or a peptide that causes ribosome skipping or self cleavage (e.g., a 2A sequence, e.g. a T2A, E2A, F2A, or P2A), such as any described elsewhere herein. The P2A amino acid sequence can be, but is not limited to, the sequence of SEQ ID NO:301. The nucleic acid sequence encoding the P2A sequence can be, but is not limited to, the nucleic acid sequence of SEQ ID NO:302, a nucleic acid sequence encoding a P2A peptide that is at least 90% identical to the amino acid sequence of SEQ ID NO:301, or a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:302.

[0147] In some embodiments, the nucleic acid encoding the Va or Vy region or a or y chain and the nucleic acid encoding the VP or V5 region or p or 5 chain can be connected via IRES sequence.

[0148] In some embodiments, the nucleic acid sequence encoding the a and P TCR chains connected via a cleavable linker sequence comprises: a nucleotide sequence of SEQ ID NO:28, 46, or 64; a nucleotide sequence having at least 90% identify to the nucleic acid sequence of SEQ ID NO:28, 46, or 64; a nucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:27, 45, or 63; or a nucleotide sequence encoding a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO:27, 45, or 63.

[0149] In some embodiments, the nucleic acid sequence encoding the a and P TCR chains connected via a cleavable linker sequence comprises: a nucleotide sequence of SEQ ID NO: 82, 100, 118, 136, 154, 172, 190, 208, 226, 244, 262, 280, or 298; a nucleotide sequence having at least 90% identify to the nucleic acid sequence of SEQ ID NO: 82, 100, 118, 136, 154, 172, 190, 208, 226, 244, 262, 280, or 298; a nucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, or 297; or a nucleotide sequence encoding a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO: 81, 99, 117, 135, 153, 171, 189, 207, 225, 243, 261, 279, or 297.

[0150] Also provided herein are vectors or constructs containing such nucleotide sequences. In some embodiments, the vectors or constructs contain one or more heterologous promoters operatively linked to the nucleotide encoding the Va or Vy region or a or y chain and / or the VP or V5 region or P or 5 chain. In some embodiments, the heterologous promoter is operatively linked to one or more than one nucleotide sequence.

[0151] In some embodiments, the vector or construct can contain a single promoter that drives the expression of one or more nucleotide sequences. In some embodiments, the promoter is a heterologous promoter. In some embodiments, such vectors or expression constructs can be multi ci str onic (e.g., bicistronic or tricistronic, see e.g., U.S. Patent No. 6,060,273). For example, in some embodiments, transcription units can be engineered as a bicistronic unit containing an IRES (internal ribosome entry site), which allows coexpression of gene products (e.g., encoding an a or y chain and / or a P or 5 chain of a TCR) by a message from a single promoter. Alternatively, in some cases, a single promoter may direct expression of an RNA that contains, in a single open reading frame (ORF), two or three genes (e.g., encoding an a or y chain and / or a P or 5 chain of a TCR) separated from one another by sequences encoding a self-cleavage peptide (e.g., a 2A peptide, e.g., a P2A peptide) or a protease recognition site (e.g., furin). An ORF can encode a single polyprotein, which, either during (in the case of 2 A e.g., P2A) or after translation, is cleaved into the individual proteins. In some embodiments, the peptide, such as P2A, can cause the ribosome to skip (ribosome skipping) synthesis of apeptide bond at the C-terminus of a 2A element, leading to separation between the end of the 2A sequence and the next peptide downstream (see, for example, de Felipe. Genetic Vaccines and Ther. 2: 13 (2004) and deFelipe et al. Traffic 5:616-626 (2004)). Examples of 2A cleavage peptides, including those that can induce ribosome skipping, include Thosea asigna virus (T2A), porcine teschovirus-1 (P2A, e.g., SEQ ID NO:301), equine rhinitis A virus (E2A), and 2A sequences from the foot-and-mouth disease virus (F2A) as described in U.S. Patent Publication No. 2007 / 0116690. In some embodiments, the peptide that causes ribosome skipping is a P2A peptide and / or contains the sequence of amino acids set forth in SEQ ID NO:301.

[0152] In a bicistronic vector, a nucleic acid sequence encoding the Va or Vy region or a or y chain and a nucleotide sequence encoding the VP or V5 region or p or 5 chain can be present in any order and are separated by a nucleotide sequence encoding a peptide sequence that causes ribosome skipping. For example, in some embodiments, the nucleotide sequence comprises, in order, a nucleic acid sequence encoding a P or 5 chain, a nucleic acid sequence encoding a peptide sequence that causes ribosome skipping (e.g., a P2A sequence as described herein), and a nucleic acid sequence that encodes an a or y chain. In other embodiments, the nucleotide sequence contains, in order, a nucleic acid sequence that encodes an a or y chain, a nucleic acid sequence that encodes a peptide sequence that causes ribosome skipping (e.g., a P2A sequence as described herein), and a nucleic acid sequence that encodes a P or 5 chain.

[0153] In some embodiments, the nucleotide sequence encoding an Va or Vy region or a or y chain and / or the nucleotide sequence encoding a VP or V5 region or P or 5 chain of a TCR comprises one or more nucleic acid sequences corresponding to SEQ ID NOs: set forth in Table 3 for TCRs A-P. Also, among the provided nucleotide sequences encoding a TCR are those containing sequences at least or about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences. In some aspects, the nucleotide sequences contain sequences encoding a signal sequence, and the encoded exemplary anti-NPMlc TCRs, when expressed as a mature protein, comprise the mature Va and / or mature VP region, or the mature Vy and / or V5 mature region, for example, without the signal sequence (e.g., from cleavage of the signal sequence) when fully processed and expressed.Table 3: SEQ ID NOs. of Nucleotide Sequences of AIQDLCLAV (SEQ ID NO: 1) Specific TCRs.

[0154] Also provided herein are vectors, such as those containing any of the nucleic acids described herein. In some embodiments, nucleic acid or nucleic acids encoding one or both chains of a TCR, are cloned or assembled into a suitable expression vector or vectors. The expression vector can be any suitable recombinant expression vector, and can be used to transform or transfect any suitable host. Suitable vectors include those designed for propagation and expansion or for expression or both, such as plasmids and viruses. In some embodiments, the vector is an expression vector.III. METHODS FOR ISOLATING, ASSESSING, AND IDENTIFYING T CELL RECEPTORS

[0155] In some aspects, provided herein are methods for isolating a plurality of nucleic acid sequences encoding the TCRs specific for AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5). In some aspects, AIQDLCLAV (SEQ ID NO: l)-specific or CLAVEEVSL (SEQ ID NO: 5)-specific TCRs are identified based on the methods described herein. In some aspects, the methods also include isolating nucleic acid sequences, assembling the nucleic acidsequences into vectors, assessing the expression and / or activity of the TCRs, and screening and identifying particular TCRs of interest, in some cases, using a high-throughput method.

[0156] The methods described herein also relate to determining the binding activity and functional capacity of candidate TCRs.A. TCR Donor criteria and methods for isolating NPMlc specific T cell candidates

[0157] In some embodiments, whole exome sequencing is used to type donors for their HLA repertoire and or haplotype. In some embodiments, the donor subject has cancer. In some embodiments, the donor subject has cancer of a type that is to be treated with the engineered T cells. In some embodiments, the donor is a healthy donor. In some embodiments, those subjects with an appropriate HLA type are selected for blood collection. Anti-AIQDLCLAV (SEQ ID NO: 1) reactive T cells from the donor subject(s) can be identified on the basis of their binding an HLA-multimer folded with the neoantigen AIQDLCLAV (SEQ ID NO: 1) peptide. These cells are single cell sorted and their TCRs are screened for anti -AIQDLCLAV (SEQ ID NO: 1) reactivity. In some embodiments, the donor is HLA-A*02:01+, HLA-A*02:06+, or HLA- A*02:03+. Anti-CLAVEEVSL (SEQ ID NO: 5) reactive T cells from the donor subject(s) can be identified on the basis of their binding an HLA-multimer folded with the neoantigen CLAVEEVSL (SEQ ID NO: 5) peptide. These cells are single cell sorted and their TCRs are screened for anti -CLAVEEVSL (SEQ ID NO: 5) reactivity. In some embodiments, the donor is HLA-A*02:01+, HLA-A*02:06+, or HLA-A*02:03+. Reactive T cells can also be identified by proliferation or expansion, cytokine expression, upregulation of activation / signaling proteins, upregulation or downregulation of other proteins metabolites or other cellular constituents, changes in cell shape or granularity, or other means available in the art.B. High-Throughput Isolation, Amplification and Assembly of Nucleic AcidSequences Encoding TCRs

[0158] In some aspects, nucleic acid molecules encoding a TCR can be obtained or identified from a variety of sources. In some aspects, TCRs can be obtained or identified using a high- throughput TCR isolation and screening method. Examples of such methods that can be used include those described in, for example, WO2018 / 102473, which is incorporated herein by reference in its entirety. In some aspects, the high-throughput TCR isolation and screening methods involve the amplification of nucleic acids encoding TCR a and / or P chains, or TCR y and / or 5 chains, from a plurality of different cells, such as T cells, isolated from a donor. Also provided herein are such methods related to isolating or screening a plurality of different TCRs to obtain TCRs that are specific for an NPMlc neoantigen, such as AIQDLCLAV (SEQ IDNO: 1) or CLAVEEVSL (SEQ ID NO: 5).

[0159] In some embodiments, nucleic acid molecules encoding a TCR can be obtained from a variety of sources, such as by polymerase chain reaction (PCR) amplification of encoding nucleic acids within or isolated from a given cell or cells. In some embodiments, a TCR is obtained from a biological source, such as T cells (e.g., cytotoxic T cells), T cell hybridomas, or other publicly available sources. In some embodiments, a TCR may be derived from one of various animal species, such as a human, mouse, rat, or other mammal. In some embodiments, the T cells can be obtained from in vivo isolated cells, such as from normal (or healthy) subjects or diseased subjects, including T cells present in peripheral blood mononuclear cells (PBMCs) or tumor-infiltrating lymphocytes (TILs). In some embodiments, the T cells can be a cultured T cell hybridoma or clone. For example, in some embodiments, to generate a vector encoding a TCR, the a and P chains can be PCR amplified from total cDNA isolated from a T cell clone expressing the TCR of interest and cloned into an expression vector. In some embodiments, the a and P chains can be synthetically generated. In some embodiments, the a and P chains are cloned into the same vector.

[0160] As described herein, the methods and materials provided herein can allow users to capture successfully most, if not all, functional TCRs from a sorted T cell population. For example, an amplification (e.g., nested amplification procedure such as a nested PCR) procedure can include using primer collections designed to amplify every known functional V segment, or nearly every (>90%, >95%, >96%, >97%, or >98%) known V segment of the two variable chains of a particular TCR (e.g., any of the known functional V segments of the a variable and P variable chains of a particular aP TCR or any of the known functional V segments of the y variable and 5 variable chains of a particular y5 TCR) of a mammal (e.g., a human). For humans, an amplification procedure can include a primer collection designed to amplify all or nearly all (>90%, >95%, >96%, >97%, or >98%) 45 V segments of the a chain currently known to be functional and all or nearly all (>90%, >95%, >96%, >97%, or >98%) 48 V segments of the P chain currently known to be functional. When referring to TCR V segments of the a chain herein, the shorthand abbreviation TRAV can be used. Likewise, when referring to TCR V segments of the P chain herein, the shorthand abbreviation TRBV can be used. The same is true for TCR V segments of the y and 5 chains, which can be referred to as TRGV and TRDV, respectively.

[0161] In some aspects, this document provides methods and materials involved in cloning functional TCRs from single T cells. For example, in some aspects, this document provides methods and materials for obtaining nucleic acid encoding a TCR from a single T cell andarranging that nucleic acid to form nucleic acid vectors successfully designed to express a TCR (e.g., a fully intact TCR such as a fully intact TCR having the variable chain combination as present in that single T cell), kits for obtaining nucleic acid encoding a TCR from a single T cell and arranging that nucleic acid to form nucleic acid vectors successfully designed to express a TCR (e.g., a fully intact TCR such as a fully intact TCR having the variable chain combination as present in that single T cell), and methods for making such kits. A cloned aP TCR having the variable chain combination as present in a single T cell used to clone that TCR can include the VJ a segment combination as present in that single T cell, the VDJ p segment combination as present in that single T cell, the nucleotide sequence of the entire a variable region as present in that single T cell, and the nucleotide sequence of the entire P variable region as present in that single T cell. Likewise, a cloned y5 TCR having the variable chain combination as present in a single T cell used to clone that TCR can include the VJ y segment combination as present in that single T cell, the VDJ 5 segment combination as present in that single T cell, the nucleotide sequence of the entire y variable region as present in that single T cell, and the nucleotide sequence of the entire 5 variable region as present in that single T cell.

[0162] In some aspects, this document provides collections of nucleic acid primers designed to amplify the entire coding sequence of both variable regions (e.g., the a variable region and P variable region, or the y variable region and 5 variable region) for each expressed V segment (e.g., each expressed a V segment and P V segment, or each expressed y V segment and 5 V segment) for functional aP or y5 TCRs of a particular mammalian species (e.g., a mouse or a human), methods for using such collections of nucleic acid primers to clone functional TCRs from single T cells, and kits containing such collections of nucleic acid primers to clone functional TCRs from single T cells.

[0163] In some aspects, the methods and materials provided herein can allow one to perform highly multiplexed reactions to clone many different TCRs (e.g., hundreds to thousands or more different TCRs) directly from single T cells quickly (e.g., simultaneously in some cases) and in a manner that misses few, if any, a / p variable chain combinations (or y / 5 variable chain combinations). For example, the methods and materials provided herein can be performed to clone many different aP TCRs (e.g., hundreds to thousands or more different aP TCRs) directly from single aP T cells in a manner that misses less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the a variable chains and less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent,or less than 1 percent) of the P variable chains possible for a / p variable chain combinations of a species (e.g., mice or human species). Likewise, the methods and materials provided herein can be performed to clone many different y5 TCRs (e.g., hundreds to thousands or more different y5 TCRs) directly from single y5 T cells in a manner that misses less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the y variable chains and less than 10 percent (e.g., less than 9 percent, less than 8 percent, less than 7 percent, less than 6 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent) of the 5 variable chains possible for y / 5 variable chain combinations of a species (e.g., mice or human species). In some embodiments, the methods and materials provided herein can include (a) obtaining a sample of T cells, (b) sorting those T cells into isolated locations (e.g., wells) such that most, if not all, isolated locations (e.g., each well) contain a single T cell, (c) lysing (e.g., simultaneously lysing) the single T cells located in separate isolated locations (e.g., separate wells) to release the RNA of each single T cell, (d) performing (e.g., simultaneously performing) reverse transcription using the released RNA as template, appropriate primers for cDNA synthesis from RNA, and a reverse transcriptase enzyme to produce cDNA within each isolated location (e.g., each well); that cDNA representing the RNA expressed by the single T cell that was located in that isolated location (e.g., well), (e) performing (e.g., simultaneously performing), for each isolated location, a first round amplification reaction (e.g., a first round polymerase chain reaction (PCR)) of an amplification procedure (e.g., such as a nested amplification procedure such as a nested PCR) using the produced cDNA as template, a first round primer collection (e.g., a first round PCR primer collection), and a polymerase (e.g., Taq polymerase) to produce at least an amplification product containing a nucleic acid sequence of the a variable chain (or y variable chain) of the TCR of the single T cell of that isolated location and an amplification product containing a nucleic acid sequence of the P variable chain (or 5 variable chain) of the TCR of that same single T cell of that same isolated location, (f) performing (e.g., simultaneously performing), for each isolated location, a second round amplification reaction (e.g., a second round PCR) of a nested amplification procedure (e.g., a nested PCR procedure) using the amplification products of the first round amplification reaction as template, a second round primer collection (e.g., a second round PCR primer collection), and a polymerase (e.g., Taq polymerase) to produce at least a first amplification product containing a nucleic acid sequence of the a variable chain (or y variable chain) of the TCR of the single T cell of that isolated location and a second amplification product containing a nucleic acid sequence of the P variablechain (or 5 variable chain) of the TCR of that same single T cell of that same isolated location, and (g) cloning, for each isolated location, the first and second amplification products into an expression vector designed to express a functional TCR having the a / p or y / 5 variable chain combination (or a portion thereof such as the V segments of the a / p or y / 5 variable chain combination) as was present in the single T cell used to generate the amplification products.

[0164] The resulting expression vectors can be introduced into cells such that those cells express the cloned TCRs. Such cells and / or the TCRs they express from the introduced expression vectors can be screened to identify TCRs with desired capabilities. For example, cells expressing cloned TCRs that recognize particular antigens (e.g., NPMlc neoantigen) can be identified, and those cells, the TCR expression vectors they contain, or the cloned TCR constructs can be used for further analysis or for therapeutic applications.

[0165] In some embodiments, expression of cloned TCRs on the surface and expression of functional TCRs can be assessed by introducing the expression vectors into TCR-negative reporter cells designed to express a measurable marker signal or marker polypeptide once the signaling apparatus of a functional TCR is engaged. In these cases, an antibody designed to non-specifically activate TCRs (e.g. , an anti-CD3 antibody) can be used to screen for functional TCRs. In some embodiments, the cloned TCRs can be screened for antigen specificity. For example, reporter cells expressing cloned TCRs can be screened for the recognition of particular antigens (e.g., peptides derived from tumor polypeptides). In some embodiments, primary T cells (e.g., human primary T cells) can be transfected with expression vectors and screened for antigen specificity via T cell proliferation assays.

[0166] For example, in some cases, the methods and materials provided herein can include a nested amplification procedure (e.g., a nested PCR procedure) that includes primer collections designed to amplify every known functional V segment of the two variable chains of a particular TCR (e.g., any of the known functional V segments of the a variable and P variable chains of a particular aP TCR or any of the known functional V segments of the y variable and 5 variable chains of a particular y5 TCR) of a mammal (e.g., a human).

[0167] In some embodiments, T cells can be stimulated (e.g., in vitro stimulated) prior to sorting, and then RNA expression can be assessed (via, e.g., qPCR) to determine which T cells responded to the stimulation. Any appropriate type of stimulation can be used including, without limitation, non-specific stimulation such as stimulation with concanavalin A, phytohemagglutinin-P, phorbol esters plus ionomycin, phorbol myristate acetate plus calcium ionophores, or antibodies having the ability to cross link TCRs (e.g., anti-CD3 antibodies plus anti-CD28 antibodies, or anti-TCR P antibodies) or antigen-specific stimulation such asstimulation with one or more particular antigens as described elsewhere (Downward et al., Nature, 346:719-23 (1990); and Dasgupta et al., Proc. Natl. Acad. Set. USA, 84: 1094-8 (1987)). In some embodiments, cytokine expression levels such as TNF-a, IFN-y, IL-2, IL-4, IL-5, IL-10, IL-13, or IL-17 expression levels can be determined and compared to nonstimulated populations. Once single T cells are sorted, the methods provided herein can be used to determine which T cells were making particular cytokines in response to the stimulation (e.g., in response to a peptide antigen used to stimulate the T cells). In these cases, antigen specific T cells can be determined without laborious methods of expanding reactive T cells or the destructive methods of paraformaldehyde fixation and intracellular cytokine staining, which can reduce the ability to clone TCRs effectively. In such cases, particular TCRs generated from active and antigen specific T cells, as opposed to inactive bystander T cells, can be quickly identified.

[0168] In some embodiments, cytokine expression levels such as TNF-a, IFN-y, IL-2, IL-4, IL-5, IL-10, IL-13, or IL-17 expression levels can be determined for the single T cells used to clone functional TCRs, thereby allowing a particular TCR to be identified based on the particular phenotype (e.g., elevated IFN-y expression) of the T cell that provided the variable chains (or portions thereof) of that particular TCR. In such cases, particular TCRs generated from active, as opposed to inactive, T cells can be quickly identified. In some embodiments, particular TCRs generated from inactive, as opposed to active, T cells can be quickly identified.

[0169] In some embodiments, TCRs can be obtained using T cells refractory to stimulation, and the specificity of the cloned TCR can be tested or screened in cells where canonical TCR signaling is not repressed.

[0170] In some embodiments, an MHC -peptide complex (or an HLA-peptide complex) can be used to identify cloned TCRs that recognize such a complex. In these cases, it is possible that clonal exclusion during an immune response and / or a lack of antigen priming may result in TCRs with this specificity not being present in the activated and / or expanded TCR pool. In such cases, the methods and materials provided herein, which in some cases only requires a single T cell to be present, can be used to clone a naive or inactivated TCR that recognizes such a complex. In some embodiments, pools of naive T cells can be stained with MHC-peptide tetramers (or HLA-peptide tetramers), and any MHC-peptide (or HLA-peptide) responsive TCRs among the naive T cells can be used to clone those TCRs using the methods and materials provided herein.

[0171] In some aspects, the methods provided herein include methods for obtaining a plurality of nucleic acid vectors containing nucleic acid encoding functional T cell receptors.The method comprises, or consists essentially of, (a) obtaining a device comprising a plurality of separate locations, wherein each of the separate locations contains cDNA generated from RNA obtained from a single T cell that was sorted into the separate locations, (b) performing a nested amplification procedure using the cDNA of each of the plurality of separate locations as template to obtain a first amplification product and a second amplification product for the cDNA of each of the plurality of separate locations, wherein the first amplification product comprises nucleic acid encoding a Va or Vy segment, and wherein the second amplification product comprises nucleic acid encoding a VP or V5 segment, and (c) assembling the first amplification product and the second amplification product for the cDNA of each of the plurality of separate locations into a nucleic acid vector to obtain an assembled nucleic acid vector for the cDNA of each of the plurality of separate locations, wherein the assembled nucleic acid vectors for the cDNA of each of the plurality of separate locations comprises nucleic acid encoding a functional T cell receptor. The plurality of nucleic acid vectors can be a plurality of nucleic acid expression vectors. The device can comprise a multi-well plate. The multi-well plate can be a 96-well plate, a 384-well plate, or a 1536-well plate. The cDNA generated from RNA obtained from a single T cell single can comprise cDNA generated from RNA obtained from a single human T cell. The first amplification product can comprise nucleic acid encoding an L sequence of a Va or Vy segment. The first amplification product can comprise nucleic acid encoding a Ja or Jy segment. The first amplification product can comprise nucleic acid encoding a 5' portion of a Ca or Cy region. The first amplification product can comprise nucleic acid encoding an L sequence of a Va or Vy segment, a Ja or Jy segment, and a 5' portion of a Ca or Cy region. The second amplification product can comprise nucleic acid encoding an L sequence of a VP or V5 segment. The second amplification product can comprise nucleic acid encoding a Dp or D5 segment. The second amplification product can comprise nucleic acid encoding a jp or J5 segment. The second amplification product can comprise nucleic acid encoding a 5' portion of a CP or C5 region. The second amplification product can comprise nucleic acid encoding an L sequence of a VP or V5 segment, a Dp or D5 segment, a jp or J5 segment, and a 5' portion of a CP or C5 region. The first amplification product can comprise an adapter sequence added to an amplified template sequence of the cDNA via a second round amplification of the nested amplification procedure. The second amplification product can comprise an adapter sequence added to an amplified template sequence of the cDNA via a second round amplification of the nested amplification procedure. The first amplification product can comprise a first adapter sequence added to an amplified template sequence of the cDNA via a second round amplification of the nested amplificationprocedure, and the second amplification product can comprise a second adapter sequence added to an amplified template sequence of the cDNA via a second round amplification of the nested amplification procedure, wherein the first and second adapter sequence are different. The functional T cell receptor of each of the assembled nucleic acid vectors can comprise a Va / VP combination or Vy / V5 combination as present in the single T cell originating the RNA. The functional T cell receptor of each of the assembled nucleic acid vectors can comprise (a) a full- length a variable region and a full-length p variable region or (b) a full-length y variable region and a full-length 5 variable region. The functional T cell receptor of each of the assembled nucleic acid vectors can comprise (a) a full-length a variable region and a full-length p variable region as present in the single T cell originating the RNA or (b) a full-length y variable region and a full-length 5 variable region as present in the single T cell originating the RNA. The functional T cell receptor of each of the assembled nucleic acid vectors can comprise (a) a full- length a constant region and a full-length P constant region or (b) a full-length y constant region and a full-length 5 constant region. Each of the assembled nucleic acid vectors can comprise a nucleic acid sequence encoding a self-cleaving peptide or an internal ribosome entry site (IRES). The method can comprise sorting single T cells into the separate locations. The method can comprise performing a reverse transcription reaction to obtain the cDNA. The assembling step can comprise seamless cloning. Each of the assembled nucleic acid vectors can be obtained without performing nucleic acid sequencing. Each of the assembled nucleic acid vectors can be obtained without performing a restriction endonuclease cleavage reaction.C. Assessing Neoantigen Specific T Cell Receptor Expression, Activity, and Function

[0172] Exemplary assays can be used to assess the activity, expression and / or function of the anti-NPMlc TCRs and antigen-binding fragments described herein. The assays described herein, which are not to be construed as limiting, may be used to assess the functional capacity of candidate NPMlc-specific TCRs.

[0173] Functional characterization of anti-NPMlc TCRs can be performed by binding assays utilizing fluorescent labeled MHC molecules carrying specific target peptides (tetramer / pentamer / dextramer), or activation assays by co-culturing anti-NPMlc TCR expressing cells with antigen presenting cells (APCs) presenting the corresponding MHC / peptide complexes.

[0174] A cytokine release assay can evaluate the ability of a candidate anti-NPMlc TCR to produce the cytokines (e.g., IL-2, TNFa, and / or IFN-y) or other T cell activation markers (e.g.,CD69) following exposure to cells presenting the target antigen. T cells are incubated with K562 cells (e.g., AIQDLCLAV (SEQ ID NO: l)(neg) / HLA-A*:02:01(pos)) loaded with the target AIQDLCLAV (SEQ ID NO: 1) peptide. As a control, K562 cells are loaded with the non-target peptide or an irrelevant peptide control. IL-2 and / or IFN-y responses of the T cells are followed by intracellular cytokine staining and analysis by FACS.

[0175] A T cell activation / degranulation marker assay can be used evaluate the ability of candidate anti-NPMlc TCRs to express the surface marker CD107a following exposure to cells presenting the target antigen. CD 107a is a marker of T cell degranulation, which is part of the cell killing response. T cells are incubated with K562 cells loaded with the target AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) peptide, or a control peptide. Degranulation responses are followed on the T cells by CD107a surface staining and analysis by FACS.

[0176] A killing assay can evaluate the ability of candidate anti-NPMlc TCRs to lyse cells presenting the target antigen. T cells are incubated with a mixture of fluorescent-tag labeled K562 cells differentially loaded with target and control peptides to allow on-target and off- target cytotoxicity to be examined within a single test sample. Fluorescent cell counting beads are included as a normalization / count control. Fluorescently tagged K562 cells are loaded with the target AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) peptide. As a control, K562 cells labeled with a different fluorescent-tag are loaded with the non-target peptide control. Gated cell counts of AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) peptide loaded K562 cells and control peptide loaded K562 cells remaining after incubation with T cells are followed by FACS.

[0177] The CD34 marker may be used as a surrogate potency measurement. See Philip et al. (2014) / c 124(8): 1277-1287. Detection of CD34, a marker of transduction efficiency, may correlate with the functional potency measurements described herein.

[0178] In some aspects, expanded and unexpanded screens are performed using the exact same donor for a direct comparison of methods. With certain methods, expansion may be performed. A method that yields candidate anti-NPMlc TCRs without expansion processes can be advantageous in some contexts in view of the time sensitivity of screening for anti- NPMlc TCRs with desired specificity. Reduced sample processing may also offer advantages in different contexts.IV. ENGINEERED CELLS

[0179] Also provided herein are cells such as cells that have been engineered to contain or express any of the anti-NPMlc TCRs described herein. Also provided herein are populationsof such cells and compositions containing such cells and / or enriched for such cells, such as in which cells expressing the anti-NPMlc TCR make up at least 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more percent of the total cells in the composition. In some embodiments, the cells are primary T cells or cells of a certain type such as T cells or CD8+or CD4+cells. Among the compositions are pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy. Also provided herein are therapeutic methods for administering the cells and compositions to subjects, e.g., patients.

[0180] Thus, also provided herein are genetically engineered cells expressing an anti-NPMlc TCR provided herein. The cells generally are eukaryotic cells, such as mammalian cells, and typically are human cells. In some embodiments, the cells are derived from the blood, bone marrow, lymph, or lymphoid organs, and are cells of the immune system, such as cells of the innate or adaptive immune system, e.g., myeloid or lymphoid cells, including lymphocytes, typically T cells and / or NK cells. Other exemplary cells include stem cells, such as multipotent and pluripotent stem cells, including induced pluripotent stem cells (iPSCs), Haemopoietic Stem Cells (HSC), or Haemopoietic Progenitor Cells (HPC). The cells typically are primary cells (e.g., primary T cells), such as those isolated directly from a subject and / or isolated from a subject and frozen. In some embodiments, the cells include one or more subsets of T cells or other cell types, such as whole T cell populations, CD4+ cells, CD8+ cells, and subpopulations thereof, such as those defined by function, activation state, maturity, potential for differentiation, expansion, recirculation, localization, and / or persistence capacities, antigenspecificity, type of antigen receptor, presence in a particular organ or compartment, marker or cytokine secretion profile, and / or degree of differentiation. With reference to the subject to be treated, the cells may be allogeneic and / or autologous. In some embodiments, the methods provided herein include isolating cells from the subject, preparing, processing, culturing, and / or engineering them, as described herein, and re-introducing them into the same patient, before or after cryopreservation.

[0181] Among the sub-types and subpopulations of T cells (including primary T cells) and / or of CD4+and / or of CD8+T cells (including primary CD4+and / or of CD8+T cells) included herein are naive T (TN) cells, effector T cells (TEFF), memory T cells and sub-types thereof, such as stem cell memory T (TSCM), central memory T (TCM), effector memory T (TEM), or terminally differentiated effector memory T cells, tumor-infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosa-associated invariant T (MAIT) cells, naturally occurring and adaptive regulatory T (Treg) cells, helper T cells, suchas TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, a / p T cells, and 8 / y T cells.

[0182] In some embodiments, the cells are NK cells or memory NK cells. In some embodiments, the cells are monocytes or granulocytes, e.g., myeloid cells, macrophages, neutrophils, dendritic cells, mast cells, eosinophils, and / or basophils.

[0183] In some embodiments, the cells include one or more nucleic acids introduced via genetic engineering, and thereby express recombinant or genetically engineered products of such nucleic acids. In some embodiments, the nucleic acids are heterologous, z.e., normally not present in a cell or sample obtained from the cell, such as one obtained from another organism or cell, which for example, is not ordinarily found in the cell being engineered and / or an organism from which such cell is derived. In some embodiments, the nucleic acids are not naturally occurring, such as a nucleic acid not found in nature, including one comprising chimeric combinations of nucleic acids encoding various domains from multiple different cell types.

[0184] In some embodiments, the expression of the endogenous TCR chains of the engineered cell is reduced or eliminated. Exemplary methods for reducing or preventing endogenous TCR expression are described, see e.g., U.S. Patent No. 9,273,283; U.S. publication no. US2014 / 0301990.

[0185] In some embodiments, a nucleic acid encoding an anti-AIQDLCLAV or anti- CLAVEEVSL CAR is transfected into a T cell to form an anti-AIQDLCLAV or anti- CLAVEEVSL CAR T cell. The T cell can be, but is not limited to, a primary T cell, a culture T cell, a autologous T cell, an allogeneic T cell, a T cell obtained from bone marrow, a T cell obtained from a lymph node, a T cell obtained from a thymus, a tumor infiltrating lymphocyte, a T cell obtained from a spleen, a T cell from umbilical cord blood, a universal allogenic T cell, a universal CAR T cell, a CAR T cell, a naive T cell, an effector T cell, an effector memory T cell, a CD4+ / CD8+ T cell, a helper T cell, a CD4+ T cell, a CD4+ helper T cell, a Thl T cell, a Th2 T cell, a Th3 cell, a Th9 cell, a tH17cell, a Th22 cell, a cytotoxic T cell, a CD8+ T cell, peripheral blood mononuclear cell (PBMC), a peripheral blood leukocyte (PBL), a memory T cell, a central memory T cell, a regulatory T cell, an aP T cell, a y5 T cell, a modified T cell, a T cell for use in adoptive cell transfer therapy, or a TCR-engineered T cell. The CAR T cell can be a first generation CAR T cell, a second generation CAR T cell, a third generation CAR T cell, a fourth generation CAR T cell, dual-antigen receptor CAR T cell, or a CAR T cell having an inducible suicide gene, or a combination thereof.A. Preparation of cells for genetic engineering

[0186] In some embodiments, preparation of the engineered cells includes one or more culture and / or preparation steps. The cells for introduction of the anti-NPMlc TCR may be isolated from a sample, such as a biological sample, e.g., one obtained from or derived from a subject. In some embodiments, the subject from which the cell is isolated (donor) is one having the disease or condition or in need of a cell therapy or to which cell therapy will be administered (i.e., autologous T cells). In some embodiments, the donor is a healthy donor. The subject in some embodiments is a human in need of a particular therapeutic intervention, such as the adoptive cell therapy for which cells are being isolated, processed, and / or engineered.

[0187] The samples include tissue, fluid, and other samples taken directly from the subject, as well as samples resulting from one or more processing steps, such as separation, centrifugation, genetic engineering (e.g., transduction with viral vector), washing, and / or incubation. The biological sample can be a sample obtained directly from a biological source or a sample that is processed. Biological samples include, but are not limited to, biopsy, tissue sample, body fluids, such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine and sweat, tissue and organ samples, including processed samples derived therefrom.

[0188] In some aspects, the sample from which the cells are derived or isolated is blood or a blood-derived sample, or is or is derived from an apheresis or leukapheresis product. Exemplary samples include whole blood, PBMCs, leukocytes, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut associated lymphoid tissue, mucosa associated lymphoid tissue, spleen, other lymphoid tissues, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testes, ovaries, tonsil, or other organ, and / or cells derived therefrom. Samples include, in the context of cell therapy, e.g., adoptive cell therapy, samples from autologous and allogeneic sources.

[0189] In some embodiments, the cells are derived from cell lines, e.g., T cell lines. The cells in some embodiments are obtained from a xenogeneic source, for example, from mouse, rat, non-human primate, or pig.B. Vectors and methods for genetic engineering

[0190] Also provided herein are methods, nucleic acids, compositions, and kits for expressing an anti-NPMlc TCR or antigen-binding fragment thereof provided herein, and for producing the genetically engineered cells expressing such anti-NPMlc TCR or antigenbinding fragment thereof. The genetic engineering generally involves introduction of a nucleic acid encoding the anti-NPMlc TCR (or antigen-binding fragment thereof) into the cell, suchas by retroviral transduction, nanoparticle delivery, transfection, or transformation.

[0191] In some embodiments, gene transfer is accomplished by first stimulating the cell, such as by combining it with a stimulus that induces a response such as proliferation, survival, and / or activation — e.g., as measured by expression of a cytokine or activation marker — followed by transduction of the activated cells, and expansion in culture to numbers sufficient for clinical applications.

[0192] Various methods for the introduction of genetically engineered components are well known and may be used with the provided methods and compositions. Exemplary methods include those for transfer of nucleic acids encoding an anti-NPMlc TCR or antigen-binding fragment thereof provided herein, including via viral, e.g., retroviral or lentiviral, transduction, transposons, electroporation, and nucleofection.

[0193] In some embodiments, recombinant nucleic acids are transferred into cells using recombinant infectious virus particles. In some embodiments, recombinant nucleic acids are transferred into T cells using recombinant lentiviral vectors or retroviral vectors, such as y- retroviral vectors.

[0194] In some embodiments, the retroviral vector has a long terminal repeat sequence (LTR), e.g., a retroviral vector derived from the Moloney murine leukemia virus (MoMLV), myeloproliferative sarcoma virus (MPSV), murine embryonic stem cell virus (MESV), murine stem cell virus (MSCV), or spleen focus forming virus (SFFV). Most retroviral vectors are derived from murine retroviruses. In some embodiments, the retroviruses include those derived from any avian or mammalian cell source. The retroviruses typically are amphotropic, meaning that they are capable of infecting host cells of several species, including humans. In some embodiments, the nucleic acid to be expressed replaces the retroviral gag, pol and / or env sequences. A number of illustrative retroviral systems have been described elsewhere (see, e.g. , U.S. Pat. Nos. 5,219,740; 6,207,453; 5,219,740; Miller and Rosman (1989) BioTechniques 7:980-990; Miller, A. D. (1990) Human Gene Therapy 1 :5-14; Scarpa et al. (1991) Virology 180:849-852; Bums et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037; and Boris-Lawrie and Temin (1993) Cur. Opin. Genet. Develop. 3: 102-109).

[0195] Methods of lentiviral transduction are known. Exemplary methods are described in, e.g, Wang et al. (2012) J. Immunother. 35(9): 689-701; Cooper et al. (2003) Blood. 101 : 1637- 1644; Verhoeyen et al. (2009) Methods Mol Biol. 506: 97-114; and Cavalieri et al. (2003) Blood. 102(2): 497-505.

[0196] In some embodiments, recombinant nucleic acids are transferred into T cells via electroporation or nucleofection (see, e.g, Chicaybam et al, (2013) PLoS ONE 8(3): e60298and Van Tedeloo et al. (2000) Gene Therapy 7(16): 1431-1437). In some embodiments, recombinant nucleic acids are transferred into T cells via transposition (see, e.g., Manuri et al. (2010) Hum Gene Ther 21(4): 427-437; Sharma et al. (2013) Molec Ther Nucl Acids 2, e74; and Huang et al. (2009) Methods Mol Biol 506: 115-126). Other methods of introducing and expressing nucleic acid provided herein in immune cells include calcium phosphate transfection (e.g., as described in Current Protocols in Molecular Biology, John Wiley & Sons, New York. N.Y.), protoplast fusion, cationic liposome-mediated transfection, tungsten particle-facilitated microparticle bombardment (Johnston, Nature, 346: 776-777 (1990)), and strontium phosphate DNA co-precipitation (Brash et al., Mol. Cell Biol., 7: 2031-2034 (1987)).

[0197] Other approaches and vectors for transfer of nucleic acid encoding an anti-NPMlc TCR, antigen-binding fragment thereof, or recombinant product provided herein include those described elsewhere. See, e.g., International Patent Application Publication No. WO2014 / 055668 and U.S. Patent No. 7,446,190.

[0198] In some embodiments, one or more additional nucleic acids can be introduced into a cell concurrently with or sequentially with nucleic acid encoding an anti-NPMlc TCR or antigen-binding fragment thereof provided herein. In some embodiments, such an additional nucleic acid for introduction can be those that improve the efficacy of therapy, such as by promoting viability and / or function of transferred cells; those that provide a genetic marker for selection and / or evaluation of the cells, such as to assess in vivo survival or localization; and / or those that improve safety, for example, by making the cell susceptible to negative selection in vivo as described (Lupton SD et al., Mol. and Cell Biol., 11 :6 (1991); Riddell et al., Human Gene Therapy 3:319-338 (1992)); PCT / US91 / 08442 and PCT / US94 / 05601 describing the use of bifunctional selectable fusion genes derived from fusing a dominant positive selectable marker with a negative selectable marker; and US Patent No. 6,040,177).

[0199] Thus, provided in some embodiments are engineered cells, such as those containing an anti-NPMlc TCR or antigen-binding fragment thereof, nucleic acid, or vector as described herein. In some aspects, the cell is produced by transducing the cell in vitro or ex vivo with a vector described herein. In some aspects, the cell is a T cell, such as a CD8+ or CD4+ T cell. In some embodiments, the anti-NPMlc TCR is heterologous to the cell.V. THERAPEUTIC AND PROPHYLACTIC METHODS AND USES

[0200] Also provided herein are methods of administering and uses, such as therapeutic and prophylactic uses, of the anti-NPMlc TCRs and antigen-binding fragments thereof provided herein and / or engineered cells expressing the anti-NPMlc TCRs or antigen-binding fragmentsthereof. Such methods and uses include therapeutic methods and uses, for example, involving administration of the molecules, cells, or compositions containing the same, to a subject having a NPMlc+cancer or condition. In some embodiments, the molecule, cell, and / or composition is administered in an effective amount to effect treatment of the NPMlc+cancer or condition. Uses include uses of the anti-NPMlc TCRs and engineered T cells in such methods and treatments, and in the preparation of a medicament in order to carry out such therapeutic methods. In some embodiments, the methods are carried out by administering the anti-NPMlc TCRs or engineered T cells, or compositions comprising the same, to the subject having, having had, or suspected of having aNPMlc+ cancer or condition. In some embodiments, the methods thereby treat the NPMlc+ cancer or condition or condition in the subject.

[0201] The described anti-AIQDLCLAV TCRs, BiTEs, and CARs and engineered cells expressing the anti-AIQDLCLAV TCRs, BiTEs, and CARs can be used in the treatment of cancer. Also described are methods of treating cancer in a subject comprising administering to the subject any of a cell, such as a T cell, expressing any of the described anti-AIQDLCLAV TCRs, BiTEs, and CARs (engineered anti-AIQDLCLAV T cells). The engineered anti- AIQDLCLAV TCR expressing T cells can be administered to a subject to induce an immune response, increase T cell infiltration of the tumor, reduce or inhibit cancer cell growth, reduce or inhibit tumor growth, reduce tumor progression, reduce tumor mass, inhibit or reduce metastasis, reduce or inhibit the development of metastatic cancer, or increase survival or prolong life of the subject.

[0202] The described anti-CLAVEEVSL TCRs, BiTEs, and CARs and engineered cells expressing the anti-CLAVEEVSL TCRs, BiTEs, and CARs can be used in the treatment of cancer. Also described are methods of treating cancer in a subject comprising administering to the subject any of a cell, such as a T cell, expressing any of the described anti-CLAVEEVSL TCRs, BiTEs, and CARs (engineered anti-CLAVEEVSL T cells). The engineered anti- CLAVEEVSL TCR expressing T cells can be administered to a subject to induce an immune response, increase T cell infiltration of the tumor, reduce or inhibit cancer cell growth, reduce or inhibit tumor growth, reduce tumor progression, reduce tumor mass, inhibit or reduce metastasis, reduce or inhibit the development of metastatic cancer, increase survival or prolong life of the subject, or reduce or attenuate an autoimmune response.

[0203] The described anti-NPMlc TCRs and T cells expressing the described anti-NPMlc TCRs can be used to kill or eliminate NPMlc expressing cancer cells. Killing or eliminating NPMlc expressing cancer cells in a subject can be used treat NPMlc+cancer. In some embodiments, the engineered T cells expressing the anti-NPMlc TCR is an engineeredautologous T cell (z.e., the T cell is derived from the subject to be treated).

[0204] Among the diseases to be treated are cancers. In some embodiments, the disease or condition to be treated is a liquid tumor. In some embodiments, the disease or condition to be treated is a hematopoietic tumor. In some embodiments, the disease or condition to be treated is NPMlc+leukemia, NPMlc+acute myeloid leukemia (AML), or NPMlc+myelodysplastic syndrome (MDS). In some embodiments, the disease of condition is AML. AML can be diagnosed according to European LeukemiaNet (ELN) criteria (e.g., Dbhner et al., 2022).

[0205] In some embodiments, the described anti-NPMlc TCRs may be isolated and administered to a subject as soluble anti-NPMlc TCRs. In some embodiments, the any of the described anti-NPMlc TCRs may be isolated and conjugated to a molecule, such as a therapeutic molecule or an antibody to from an anti-NPMlc TCR-drug conjugate. The conjugated anti-NPMlc TCRs may then be administered to a subject. In some embodiments, the therapeutic molecule is a therapeutic molecule that targets AML. Therapeutic molecules that target AML include, but are not limited to, menin-MLL inhibitors. In some embodiments, the therapeutic molecule is a therapeutic molecule anti-cancer therapeutic. The anti-cancer therapeutic can be, but is not limited to, pseudomonas exotoxin, ricin, monomethyl auristatin F, and calicheamicin. In some embodiments, the anti-NPMlc TCR is used as a surrogate for an antibody in an antibody drug conjugate.

[0206] In some embodiments, engineered T cells expressing any of the described anti- NPMlc TCRs are administered to a subject for the treatment of a NPMlc+cancer, e.g., AML. In some embodiments, engineered T cells expressing any of the described anti-NPMlc TCRs can be formulated for use as a T cell therapy for the treatment of aNPMlc+ cancer, e.g., AML. In some embodiments, the T cell therapy is an autologous T cell therapy.

[0207] In some embodiments, any of the described anti-NPMlc TCRs or engineered cells expressing the anti-NPMlc can be used or formulated for use in combination with one or more AML therapies (e.g., Menin-MLL inhibitors). The one or more AML therapies can be administered the subject administered prior, in conjunction with, or after administering the anti- NPMlc TCRs or engineered cells expressing the anti-NPMlc to the subject.

[0208] In some embodiments, the described anti-NPMlc TCRs and / or engineered T cells expressing the described TCRs are administered to a subject to kill NPMlc+cells in the subject. The subject may or may not have received SCT. The subject may or may not be scheduled to receive a SCT. The subject may or may not be eligible for SCT.

[0209] In some embodiments, the subject has received one or more prior therapies to treat the cancer. In some embodiments, the subject has relapsed following one or more priortherapies to treat the cancer. In some embodiments, the subject has relapsed following one or more induction therapies. In some embodiments, the subject has relapsed following at least two prior therapies to treat the cancer. In some embodiments, the subject is refractory to one or more prior therapies to treat the cancer. In some embodiments, the subject is refractory to one or more induction therapies. In some embodiments, the subject is refractory to at least two prior therapies to treat the cancer. In some embodiments, the subject has persistent disease following one or more prior therapies to treat the cancer. In some embodiments, the subject has persistent disease following one or more induction therapies to treat the cancer. The one or more prior therapies or induction therapies can be but are not limited to, chemotherapy, azacytidine therapy, FLT3 inhibitor therapy, IDH inhibitor therapy, BCL-2 inhibitor therapy, hedgehog pathway inhibitor therapy, radiation therapy, aSCT, alloSCT, or Menin-MLL inhibitor therapy. In some embodiments, the subject has relapsed, is refractory to, or has persistent disease following one or more consolidation therapies. In some embodiments, the subject has relapsed following SCT or alloSCT but has not received a prior T cell therapy.

[0210] In some embodiments, the subject having an NPMlc mutation is FLT3 positive. In some embodiments, the subject having an NPMlc mutation is FLT3 positive and the engineered T cells are allogeneic T cells (z.e., T cells from an HLA matched donor). In some embodiments, the subject having an NPMlc mutation is FLT3 positive, the engineered to T cells are allogeneic T cell, and the engineered T cells are administered in combination with alloSCT.

[0211] In some embodiments, the subject has an NPMlc+cancer and is not eligible for transplant (e.g., standard of care alloSCT). In some embodiments, the subject has an NPMlc+cancer (e.g., AML) and one or more co-morbidities or risk factors. Co-morbidities or risk factors include, but are not limited to, advanced age, lung disease, liver disease, kidney failure, heart failure, infection, and risk of infection. In some embodiments, the subject is 60 years of age or older, 65 years of age or older, or 70 years of age or older.

[0212] In some embodiments, the subject has an NPMlc+cancer (e.g., AML) and is ineligible for high-intensity chemotherapy. In some embodiments, the subject has relapsed following high-intensity chemotherapy with or without hematopoietic stem cell transplant (HSCT). In some embodiments, the subject has not responded to, is not responding to, or has failed to achieve complete remission (is refractory to) high-intensity chemotherapy. In some embodiments, the subject is eligible for reduced intensity chemotherapy. The reduced intensity chemotherapy can be administered prior to administration of the engineered T cells.

[0213] In some embodiments, the subject is eligible for high-intensity treatment, butconsidered to have intermediate or unfavorable risk and has not responded to or relapsed following previous high-intensity chemotherapy.

[0214] In some embodiments, the subject has relapsed following a prior hypomethylating agent and / or BCL-2 inhibitor therapy. In some embodiments, the subject has not responded, is not responding to, or has failed to achieve complete remission (is refractory to) hypomethylating agent therapy.

[0215] In some embodiments, the subject has relapsed following a stem cell transplant. In some embodiments, the subject has not response to a stem cell transplant. In some embodiments, a subject eligible for treatment with a T cell therapy using any of the described anti-NPMlc TCRs has not received a stem cell transplant. In some embodiments, a subject eligible for treatment with a T cell therapy using any of the described anti-NPMlc TCRs has not received a stem cell transplant within two months of receiving the T cell therapy.

[0216] In some embodiments, methods of treating a subject having or diagnosed with an NPMlc+cancer are described, the methods comprising administering to the subject any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein in combination with a stem cell transplant (SCT). In some embodiments, methods of treating a subject having or diagnosed with an NPMlc+cancer are described, the methods comprising administering to the subject any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein prior to administering a SCT to the subject. In some embodiments, any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein are formulated for use in combination with a stem cell transplant for treatment of a subject having or diagnosed with a NPMlc+cancer. The anti-NPMlc TCRs, antigenbinding fragment thereof, polynucleotides, vectors, engineered cells, or compositions can be formulated for administration to the subject in combination with SCT or for administration to the subject prior to SCT. The SCT can be autologous SCT (aSCT) or allogeneic SCT (alloSCT). In some embodiments, the anti-NPMlc TCR is administered as an autologous T cell therapy. The NPMlc+cancer can be, but is not limited to, AML.

[0217] In some embodiments, methods of treating a subject having or diagnosed with an NPMlc+cancer are described, the methods comprising administering to the subject any of theanti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided. In some embodiments, the methods of treating are not combined with SCT. In some embodiments, any of the anti-NPMlc TCRs or antigen-binding fragment thereof provided herein, any of the polynucleotides provided herein, any of the vectors provided herein, any of the engineered cells provided herein, or any of the compositions provided herein can be formulated for use in the absence of stem cell transplant for treatment of a subject having or diagnosed with a NPMlc+cancer. In some embodiments, the anti-NPMlc TCR is administered as an autologous T cell therapy. The NPMlc+cancer can be, but is not limited to, AML.

[0218] In some embodiments, the subject has NPMlc+leukemia. In some embodiments, the subject has NPMlc+acute myelogenous (myeloid) leukemia (AML). In some embodiments, the subject has NPMlc+has myelodysplastic syndrome.

[0219] In some embodiments, the subject has not received any prior therapies. In some embodiments, the subject has a NPMlc+leukemia and is administered a described anti- NPMlc-TCR, anti-NPMlc TCR BiTE, anti-NPMlc-drug conjugate or engineered T cell expressing an anti-NPMlc TCR as a first therapeutic treatment. In some embodiments, the subject has received one or more prior therapies to treat the cancer. In some embodiments, the subject has relapsed following one or more prior therapies to treat the cancer. In some embodiments, the subject has relapsed following one or more induction therapies. In some embodiments, the subject is refractory to one or more prior therapies to treat the cancer. In some embodiments, the subject is refractory to one or more induction therapies. In some embodiments, the subject has persistent disease following one or more prior therapies to treat the cancer. In some embodiments, the subject has persistent disease following one or more induction therapies to treat the cancer. The one or more prior therapies or induction therapies can be but are not limited to, chemotherapy, azacytidine therapy, FLT3 inhibitor therapy, IDH inhibitor therapy, BCL-1 inhibitor therapy, hedgehog pathway inhibitor therapy, radiation therapy, aSCT, or alloSCT. In some embodiments, the subject has relapsed, is refractory to, or has persistent disease following one or more consolidation therapies. In some embodiments, the subject has relapsed following SCT or alloSCT but has not received a prior T cell therapy.

[0220] In some embodiments, the subject has a NPMlc+cancer (e.g., NPMlc+AML), has received induction therapy and is in remission.

[0221] In some embodiments, the subject has been diagnosed with a NPMlc+cancer e.g., NPMlc+AML), has received induction therapy, and the provided anti-NPMlc TCRs, antigen-binding fragments thereof, polynucleotides, vectors, engineered T cells, or compositions thereof, are administered as a consolidation therapy to the subject.

[0222] In some embodiments, the subject has a NPMlc+ cancer and has a Flt3ITD mutation. In some embodiments, the subject has a NPMlc+cancer and has a Flt3ITD mutation and the provided anti-NPMlc TCRs, antigen-binding fragments thereof, polynucleotides, vectors, engineered T cells, or compositions thereof, are administered to the subject to reduce or eliminated the need for SCT.

[0223] In some embodiments, the subject has aNPMlc+cancer and does not have a Flt3ITD mutation. In some embodiments, the subject has a NPMlc+cancer, does not have a Flt3ITD mutation and the provided anti-NPMlc TCRs, antigen-binding fragments thereof, polynucleotides, vectors, engineered T cells, or compositions thereof, are administered to the subject to reduce the risk or relapse.

[0224] In some embodiments, subject diagnosed with leukemia, or suspected of having leukemia is given a genetic test to a NPM1 genetic testing The NPM1 genetic test can be performed on a sample obtained from bone marrow or peripheral blood. The NPM1 genetic test can be any test known in the art to detect the presence of an NPMlc mutation or to detect a Type A or Type D NPM1 mutation. In some embodiments, the NPM1 genetic test comprises a PCR test or a next-generation sequencing (NGS). If the genetic test indicates the subject has an NPMlc+ cancer, then the subject may be treated with engineered T cells expressing any of the described anti-NPMlc TCRs.

[0225] In some embodiments, a subject that has tested positive for an NPMlc+ cancer is screened to assess of eligibility or suitability for engineered T cell treatment. The screen can include, but is not limited to, a NPM1 testing, testing of organ function (e.g., liver and kidney function), performance status of the subject, and presence of adverse events due to prior treatments. In some embodiments, the screening is performed up to about 45 days prior to an anticipated administration of an engineered T cell therapy.

[0226] Leukapheresis is performed on the subject to collect leukocytes from the subject. In some embodiments, CD8+T-Cells are obtained from the leukapheresis product via positive selection. In some embodiments, the CD8+T-cells are activated using an activating agent. About 24 h after activation, the T cells are engineered to knock out the endogenous TCR. Knock out of the endogenous TCR can be done using CRISPR. A nucleic acid sequence (or nucleic acid sequence) encoding any of the described anti-NPMlc TCRs is inserted into the CD8+TR cells. Insertion of the nucleic acid sequence(s) encoding the anti-NPMlc TCRs can be done using any method available in the art for inserting a transgene into a T cells. Suchmethods include, but are not limited to, lentivirus vector transduction. The engineered T cells are then expanded to obtain and adequate number for treatment. Leukapheresis can be performed about 21 to about 45 days prior to an anticipated administration of an engineered T cell therapy to the subject.

[0227] Prior to T cell treatment the subject is treating to induce lymphodepletion to enable engraftment of infused engineered T cells (e.g., TCR-T cells). Lymphodepletion can be performed using any method available in the art for Lymphodepletion. Suitable lymphodepletion regimens include, but not limited to, cyclophosphamide treatment and fludarabine treatment. Lymphdepletion can be performed about 1 to about 7 days prior to an anticipated administration of an engineered T cell therapy to the subject.

[0228] For the T cell therapy, engineered T-cells are infused at a pre-determined dose. The dose can be, but is not limited to, a dose of about 0.3 * 106cells to about 1 x 107cells, about 1 x 106cells to about 1 x 107cells, or about 3 x 106cells to about 1 x 107cells. In some embodiments, the dose is about 0.3x l06cells, about 0.4x l06cells, about 0.5x l06cells, about 0.6x l06cells, about 0.7x l06cells, about 0.8x l06cells, about 0.9x l06cells, about I x lO6cells, about 2x l06cells, about 3x l06cells, about 4x l06cells, about 5x l06cells, about 6x l06cells, about 7x 106cells, about 8x 106cells, about 9x 106cells, or about 1 x 107cells.

[0229] The terms “treat,” “treatment,” and the like, mean the methods or steps taken to provide relief from, or amelioration or alleviation of the number, severity, adverse effect, and / or frequency of one or more symptoms or pathological consequences of a disease, disorder, or condition in a subject. Treatment can be prophylactic in terms of preventing or partially preventing a disease, or a symptom or condition of the disease. Preventing includes providing prophylaxis with respect to the occurrence or recurrence of a disease in a subject that may be predisposed to the disease but has not yet been diagnosed with the disease. Preventing also includes providing prophylaxis with respect to the occurrence or recurrence of a symptom or pathological consequence of a disease in a subject that may be predisposed symptom or pathological consequence of the disease but has not yet been diagnosed with the symptom or pathological consequence the disease. Treatment can also be prophylactic in terms of delaying onset of a disease, or a symptom or condition of the disease. Delaying development of a disease or symptom or pathological consequence of the disease indicates deferring, hindering, slowing, retarding, stabilizing, suppressing, and / or postponing development of the disease or symptom or pathological consequence of the disease. The delay can be of varying lengths of time, depending on the history of the disease and / or individual being treated. Treating can include inhibiting the disease, disorder or condition, e.g., impeding its progress; and relieving thedisease, disorder, or condition, e.g., causing regression of the disease, disorder and / or condition. Treatment can also mean prolonging survival as compared to expected survival in the absence of treatment. Treatment can be therapeutic in terms of a partial or complete cure of a disease, condition, symptom or adverse effect attributed to the disease, disorder, or condition. The term treatment can include: (a) preventing the disease from occurring in a subject who may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., mitigating or ameliorating the disease and / or its symptoms or conditions. Treating can refer to both therapeutic treatment alone, prophylactic treatment alone, or both therapeutic and prophylactic treatment. Those in need of treatment (subjects in need thereof) can include those previously diagnosed with a disease, disorder, or condition, or those identified as being at risk of developing a disease, disorder, or condition. Treating the disease, disorder, or condition can include ameliorating at least one symptom of the particular disease, disorder, or condition, even if the underlying pathophysiology is not affected.

[0230] An “effective amount” of an agent, e.g., a pharmaceutical formulation, TCR, cells, or composition, in the context of administration, refers to an amount effective, at dosages / amounts and for periods of time necessary, to achieve a desired result, such as a therapeutic or prophylactic result.

[0231] A “therapeutically effective amount” of an agent, e.g., a pharmaceutical formulation, TCR, or cells, refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result, such as for treatment of a disease, condition, or disorder, and / or pharmacokinetic or pharmacodynamic effect of the treatment. The therapeutically effective amount may vary according to factors such as the disease state, age, sex, and weight of the subject, and the populations of cells administered. In some embodiments, the provided methods involve administering the TCRs, cells, and / or compositions at effective amounts, e.g., therapeutically effective amounts.

[0232] As used herein, a “subject” is a mammal, such as a human or other animal, and typically is human.VI. DEFINITIONS

[0233] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some embodiments, terms with commonly understood meanings are defined herein for clarityand / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0234] The terms “polypeptide” and “protein” are used interchangeably to refer to a polymer of amino acid residues, and are not limited to a minimum length. Polypeptides, including the provided T cell receptors, antigen binding fragments thereof and other peptides, e.g., linkers, may include amino acid residues including natural and / or non-natural amino acid residues. The terms also include post-expression modifications of the polypeptide, for example, glycosylation, sialylation, acetylation, phosphorylation, and the like. In some aspects, the polypeptides may contain modifications with respect to a native or natural sequence, as long as the protein maintains the desired activity. These modifications may be deliberate, as through site-directed mutagenesis, or may be accidental, such as through mutations of hosts which produce the proteins or errors due to PCR amplification.

[0235] An “isolated” nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0236] “An isolated nucleic acid molecule encoding a TCR” refers to a single nucleic acid molecule (e.g., single vector) that encodes a TCR such as a functional a / p TCR or a functional y / 5 TCR.

[0237] “An isolated nucleic acid molecule encoding an antigen binding fragment of a TCR” refers to a single nucleic acid molecule (e.g., single vector) that encodes an antigen binding fragment of a TCR.

[0238] “Isolated nucleic acid molecules encoding a TCR” refers to two or more separate nucleic acid molecules (e.g., two or more vectors) that together encode a TCR such as a functional a / p TCR or a functional y / 8 TCR. Each of such two or more nucleic acid molecules can be present at different locations within a host cell.

[0239] “Isolated nucleic acid molecules encoding an antigen binding fragment of a TCR” refers to two or more nucleic acid molecules (e.g., two or more vectors) that together encode an antigen binding fragment of a TCR. Each of such two or more nucleic acid molecules can be present at different locations within a host cell.

[0240] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include “transformants” and “transformed cells,”which include the primary transformed cell and progeny derived therefrom without regard to the number of passages. Progeny may not be completely identical in nucleic acid content to a parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0241] As used herein, “percent (%) amino acid sequence identity” and “percent identity” when used with respect to an amino acid sequence (reference polypeptide sequence) is defined as the percentage of amino acid residues in a candidate sequence (e.g., the subject T cell receptor or fragment) that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

[0242] An amino acid substitution may include replacement of one amino acid in a polypeptide with another amino acid. Amino acid substitutions may be introduced into a TCR or antigen binding fragment thereof, of interest and the products screened for a desired activity, e.g., retained / improved antigen binding, decreased immunogenicity, or improved cytolytic activity.

[0243] Amino acids generally can be grouped according to the following common side-chain properties:(1) hydrophobic: Norleucine, Met, Ala, Vai, Leu, He;(2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gin;(3) acidic: Asp, Glu;(4) basic: His, Lys, Arg;(5) residues that influence chain orientation: Gly, Pro; and(6) aromatic: Trp, Tyr, Phe.

[0244] In some embodiments, conservative substitutions can involve the exchange of a member of one of these classes for another member of the same class. In some embodiments, non-conservative amino acid substitutions can involve exchanging a member of one of these classes for another class.

[0245] The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors.”

[0246] As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. For example, “a” or “an” means “at least one” or “one or more.” It is understood that aspects and variations described herein include “consisting of’ and / or “consisting essentially of’ aspects and variations.

[0247] Throughout this disclosure, various aspects of the claimed subject matter are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the claimed subject matter. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, where a range of values is provided, it is understood that each intervening value, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the claimed subject matter. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the claimed subject matter, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the claimed subject matter. This applies regardless of the breadth of the range.

[0248] As used herein, the terms “about” and “approximately,” when used to modify an amount specified in a numeric value or range, indicates the numeric value as well as reasonable deviations from the value known to the skilled person in the art. In some embodiments, the term “about” means within the typical ranges of tolerances in the art. In some embodiments, the term “about” means within 1 or 2 standard deviations from the mean. In some embodiments, the term “about” means ±10%. In some embodiments, the term “about” means ±5%. When the term “about” is present before a series of numbers or a range, it is understood that “about” can modify each of the numbers in the series or range.

[0249] As used herein, a composition refers to any mixture of two or more products, substances, or compounds, including cells. It may be a solution, a suspension, liquid, powder, a paste, aqueous, non-aqueous or any combination thereof.

[0250] As used herein, a statement that a cell or population of cells is “positive” for a particular marker refers to the detectable presence on or in the cell of a particular marker, typically a surface marker. When referring to a surface marker, the term refers to the presence of surface expression as detected by flow cytometry, for example, by staining with an antibody that specifically binds to the marker and detecting said antibody, wherein the staining is detectable by flow cytometry at a level substantially above the staining detected carrying out the same procedure with an isotype-matched control under otherwise identical conditions and / or at a level substantially similar to that for cell known to be positive for the marker, and / or at a level substantially higher than that for a cell known to be negative for the marker.

[0251] As used herein, a statement that a cell or population of cells is “negative” for a particular marker refers to the absence of substantial detectable presence on or in the cell of a particular marker, typically a surface marker. When referring to a surface marker, the term refers to the absence of surface expression as detected by flow cytometry, for example, by staining with an antibody that specifically binds to the marker and detecting said antibody, wherein the staining is not detected by flow cytometry at a level substantially above the staining detected carrying out the same procedure with an isotype-matched control under otherwise identical conditions, and / or at a level substantially lower than that for cell known to be positive for the marker, and / or at a level substantially similar as compared to that for a cell known to be negative for the marker.

[0252] The term “specifically binds” to a target when referring to a binding region (e.g. , TCR binding region) refers to a binding reaction whereby the binding region binds to the target with greater affinity, greater avidity, and / or greater duration than it binds to a structurally different target. A binding region may have at least 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20- fold, 25-fold, 50-fold, 100-fold, 1,000-fold, 10,000-fold, or greater affinity for a specific target compared to an unrelated target when assayed under the same affinity assay conditions.EXAMPLES

[0253] The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention. CDR3PExample 1. Exemplary Identified TCRs.

[0254] Table 4 lists the sequence identifiers (SEQ ID NOs) for amino acid (aa) or nucleotide (nt) sequences for AIQDLCLAV (SEQ ID NO:1) and CLAVEEVSL (SEQ ID NO:5) specific TCRs (i.e., anti-AIQDLCLAV TCRs and anti-CLAVEEVSL TCRs) that were isolated, assessed, and sequenced. The table also lists the sequence identifier (SEQ ID NOs)corresponding to an exemplary full-length, including the constant domains, amino acid sequence containing the a and P chain sequences of each respective TCR, separated by a sequence encoding a ribosome-skip P2A sequence (P2A linker set forth in SEQ ID NO: 301 encoded by the nucleotides set forth in SEQ ID NO:302) (designated “a-P2A-P”).Table 4: Amino Acid and Nucleotide Sequences (SEQ ID NOs) of AIQDLCLAV (SEQ ID NO: l)-Specific TCRsExample 2. Characterization of Exemplary AIQDLCLAV (SEQ ID NO: 1) Specific TCRs.

[0255] As described herein, AIQDLCLAV (SEQ ID NO: 1) and / or CLAVEEVSL (SEQ ID NO:5) is expressed on cancerous cells (e.g., AML cells) but is not expressed on non-cancerous cells. For this reason, T cells targeting AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) are unlikely to cause GVHD.Example 3. Binding Specificity of Exemplary NPM1 Specific TCR

[0256] Binding specificity of the AIQDLCLAV (SEQ ID NO: 1) and CLAVEEVSL (SEQ ID NO: 5) specific TCRs was determined using tetramers complexed with AIQDLCLAV (SEQ ID NO: 1), CLAVEEVSL (SEQ ID NO:5), or control peptides.

[0257] HEK293 suspension cells were engineered to express human CD3y, CD35, CD3s, CD3(^ (polyCD3) and human CD8 a / p. CD3 and CD8 are cloned from pooled PBMCs from two healthy blood donors and stably transduced into HEK293. Suspension HEK293-CD3-CD8 cells were transiently transfected with plasmid DNA containing an anti-AIQDLCLAV TCR or an anti-CLAVEEVSL TCR and the fluorescent protein mCherry, as a transfection efficiency control.

[0258] Twenty-four hours after transfection, cells were stained with an amine-reactive viability dye (violet 510 Ghost dye), anti-CD3 and the immunogenic AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5) peptide tetramer or control peptide tetramer. Cells were acquired and analyzed by flow cytometry.

[0259] Double positive fluorescence of CD3 and specific binding of HLA-tetramers complexed with the AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) peptide indicate a functional reactive TCR.Example 4. In vitro Evaluation of Engineered T Cell Reactivity to AIQDLCLA V (SEQ ID NO:1).

[0260] TCRs isolated and identified as described in Examples 1-3 above were recombinantly expressed in a Jurkat cell line engineered to stably express CD8, and further characterized and assessed for function, including ECso determination. Cytokine secretion, T cell activation and binding specificity can also be determined for each of the TCRs.

[0261] For an exemplary AIQDLCLAV (SEQ ID NO: 1)- or CLAVEEVSL (SEQ ID Nonspecific TCR, Jurkat T cells expressing the anti-AIQDLCLAV TCR or anti-CLAVEEVSL TCR were co-cultured with K562cells pulsed with AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5) peptide or a control peptide. Equal numbers of Jurkat T cells and K562cells loaded with increasing concentrations (e.g., 0.1-31.6 ng / ml) of either AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5) peptide or control peptide were co-cultured for 16 hours. Jurkat CD69 expression was determined by FACS.A. Assessment of Early Activation Marker CD69

[0262] Expression of CD69, a marker of T cell activation and function, was assessed following co-culture of Jurkat T cells expressing an anti-AIQDLCLAV TCR or anti- CLAVEEVSL TCR with APCs loaded with AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5) peptides.

[0263] Jurkat T cells expressing various anti-AIQDLCLAV TCRs or anti-CLAVEEVSL TCR s are prepared as described. Jurkat cells transduced with anti-AIQDLCLAV TCRs or anti-CLAVEEVSL TCRs were incubated overnight with A*02:01 LCLs at a 1: 1 E / T ratio in the presence of increasing concentrations of AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO:5) peptide. The EC50 values were calculated based on XLFit (ID Business Solutions) on plots of the percentage of CD69+cells (y-axis) vs. peptide concentration (logarithmic x-axis).

[0264] For an exemplary TCR, the anti-AIQDLCLAV TCR- or anti -CLAVEEVSL TCR- expressing T cells were co-cultured in duplicate at a 1 : 1 effector-to-target ratio with peptide- loaded K562 cells. After 16-24 hours of culture, cells were washed and stained with Ghost Dye, anti-CD3 and anti-CD69 and assessed by flow cytometry. Data were analyzed by XLFit. The estimated EC50 of each of the five experimental runs was determined by assessing the ratio of CD69 positive Jurkat cells to total live cells.B. Assessment of T Cell Receptor Specificity

[0265] Specificity of the candidate anti-AIQDLCLAV or anti-CLAVEEVSL TCRs for the AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5) peptides versus corresponding wild type NPM1 peptide presented by the restricting HLA, or potentially other HLA molecules, or other HLA molecules presenting other peptides, is also assessed.2. Alloreactivity

[0266] Alloreactivity was tested utilizing HLA-A*02:01 tetramers loaded with irrelevant peptides (such as HPV peptides)Example 5. Target Cell Killing by Primary T Cells Transduced with AIQDLCLAV (SEQ ID NO:l)-Specific TCRs.

[0267] To determine the potential for inducing a graft-versus-leukemia (GvL) effect, the antileukemia target cell killing activity of primary T cells transduced with an anti-AIQDLCLAV TCR or anti-CLAVEEVSL TCRs was assessed.A. Expression of AIQDLCLAV (SEQ ID NO: 1)-Specific or CLAVEEVSL (SEQ ID NO:5)-Specific TCRs in Primary Cells

[0268] Primary human CD8+T cells were enriched from PBMCs by negative selection. The endogenous TCR of the CD8+T cells was knocked out via CRISPR. The CD8+T cells were then transduced with a lentivirus vector (pLVG.M containing an MNDU3 promoter) encoding an anti-AIQDLCLAV TCR or anti-CLAVEEVSL TCR and a RQR8 tag. The percentage of the transduced T cells was evaluated the by expression of RQR8, using an anti-CD34 antibody, by flowcytometry at day 5 post-transduction.B. Target Cell Killing

[0269] The target cell killing specificity of anti-AIQDLCLAV TCRs or anti-CLAVEEVSL TCRs was assessed. An HLA-A*02:01+ / AIQDLCLAV+(SEQ ID NO: 1) 0CI-AML2 cells or an HLA-A*02:01+ / CLAVEEVSL+(SEQ ID NO:5) OCI-AML2 cells were used as an APC. The OCI-AML2 cells were stained with Violet Proliferation Dye 450 (VPD450). VPD450 labels target cells by binding of the dye to intracellular protein, and indicates T cell-mediated target cell killing. Apoptosis of labeled cells results in the loss of their detection in the live cell gate in flow cytometry. Primary human T cells expressing an anti-AIQDLCLAV TCR or anti- CLAVEEVSL TCR were co-cultured with the LCLs for 18 hours starting at an E:T ratio (transduced CD34+positive cells) serial dilution starting at 10: 1 and ending at 0: 1. Cells were stained with 7-AAD (to evaluate the cell viability), acquired, and assessed by flow cytometry.Example 6. Target Cell Killing Activity

[0270] The target cell killing activity of an exemplary TCR was further assessed at different effectortarget (E:T) ratios.

[0271] 0CI-AML3 cells were genotyped for the AIQDLCLAV (SEQ ID NO: 1) and / or CLAVEEVSL (SEQ ID NO:5) expression. In order to distinguish cell populations, 0CI-AML3 cells presenting the AIQDLCLAV (SEQ ID NO: 1) and / or CLAVEEVSL (SEQ ID NO:5) peptide were labeled with a high concentration of VPD450 (0.5 pM), while 0CI-AML2 cells presenting the corresponding wild-type NPM1 peptide were labeled with a low concentration (0.025 pM) of VPD450. 0CI-AML2 cells and 0CI-AML3 cells were stained with VPD450 for 15 minutes at 37°C. Target (AIQDLCLAV (SEQ ID NO: 1) or CLAVEEVSL (SEQ ID NO: 5)) and control peptide-bearing OCI-AML2 cells and OCI-AML3 cells were mixed together in a 1 : 1 ratio prior to incubation with increasing numbers of transduced primary T cells expressing an AIQDLCLAV (SEQ ID NO: l)-specific or CLAVEEVSL (SEQ ID NO:5)-specific TCR. Additionally, target 0CI-AML2 or 0CI-AML3 cells were stained and incubated individually at a 1 : 1 ratio with TCR-Ts. Subsequent E:T ratio EC50 kill curves could be determined.

[0272] 0CI-AML2 cells and 0CI-AML3 cells were co-cultured with increasing numbers of primary T cells for 16 hours (E:T ratios from 1 : 16 to 10: 1). Cell mixtures were stained with LIVE / DEAD Fixable Violet Dead Cell Stain Kit (Thermo Fisher Scientific), anti-CD8 and anti-CD19, to assess the two distinct target cell populations by VPD450 staining levels. Cells were acquired and analyzed by flow cytometry, to assess CD8- cells (to exclude effector cells) and the VPD450high versus VPD4501ow populations.

[0273] Live cell counts of 0CI-AML2 cells and OCLAML3 cells presenting either target or control peptide following incubation with non-transduced T cells or anti-AIQDLCLAV TCRor anti-CLAVEEVSL TCR transduced T cells was determined. Loss of viable cells presenting target peptide (high VPD450 staining), with concomitant observation of retention of viable cells presenting control peptide (low VPD450 staining) as the effector to target (E:T) ratio was increased indicated selective killing of target cells presenting the AIQDLCLAV (SEQ ID NO: 1) and / or CLAVEEVSL (SEQ ID NO:5) by T cells genetically engineered to express an AIQDLCLAV (SEQ ID NO: l)-specific or CLAVEEVSL (SEQ ID NO:5)-specific TCR.Example 7. In Silico prediction of binding of AIQDLCLAV (SEQ ID NO:1) peptide to HLA.

[0274] Binding of AIQDLCLAV, IQDLCLAV, and AIQDLCLA to HLA was predicted using the ANN 4.0 option on IEDB (see below). Predicted IC50 values for AIQDLCLAV binding to HLA-A alleles are shown in Table 6a-d. Based on the predicted binding of AIQDLCLAV, IQDLCLAV, and AIQDLCLA, subjects having an HLA-AHLA-A*02:06, HLA-A*02:03, HLA-A*02:01 are expected to be suitable candidatesTable 6a. Predicted binding of AIQDLCLAV, IQDLCLAV, and AIQDLCLA to various HLA-A alleles.Table 6b. Predicted binding of TDQEAIQDLCLAVEEVSLRK, TDQEAIQDLCMAVEEVSLRK, TDQEAIQDLCVAVEEVSLRK to various HLA-A alleles. (8-, 9-, and 10-mers queried).Table 6b. Predicted binding of TDQEAIQDLCLAVEEVSLRK, TDQEAIQDLCMAVEEVSLRK, TDQEAIQDLCVAVEEVSLRK to various HLA-B alleles. (8-, 9-, and 10-mers queried).Table 6d. Predicted binding of TDQEAIQDLCLAVEEVSLRK, TDQEAIQDLCMAVEEVSLRK, TDQEAIQDLCVAVEEVSLRK to various HLA-C alleles. (8-, 9-, and 10-mers queried).Example 8. TCR induced signaling in engineered T cells.

[0275] TCR D was evaluated for the ability to induce T cell signaling by HLA*A02:01 presented target peptide (CLAVEEVSL, SEQ ID NO:5). In brief, Jurkat cells were engineered to express a-TCR D. K562 HLA*A02:01 expressing cells were peptide pulsed with the target peptide and subsequently co-cultured to evaluate TCR signaling. Expression of CD69 on Jurkat cells was evaluated as a surrogate marker of T cell activation by the target peptide.

[0276] Jurkat and K562 cell lines were cultured in RPMI and IMDM, respectively. Peptides were reconstituted and diluted in PBS prior to peptide pulse.

[0277] CD8+Jurkat cells were virally transduced (lentivirus) with TCR D and purified by FACS based sorting on TCR positive cells (Jurkat-TCR D cells). K562 were generated via pan HLA class I KO, virally transduced with HLA*A02:01 and purified by FACS sorting prior to performing reporter assays.

[0278] For reporter assays, engineered K562 cells were peptide pulsed with titrated concentrations of target peptide for 1 h prior to co-culture. Jurkat-TCR D cells were co-cultured with peptide pulsed K562 cells at an E:T ratio of 1 : 1 in 96 well plates. After 16-18 h, the co- cultured cells were stained with fluorescently tagged antibodies for CD3 (Jurkat specific marker), CD34 (marker for Jurkat cells transduced with the TCR of interest) and CD69 (to evaluated Jurkat-TCR D activation following co-culture). Cells were also stained with live / dead staining and analyzed utilizing NovacyteQuanteon. The ECso for peptide induced Jurkat activation was determined based on the percentage of Jurkat CD69 positive cells.

[0279] Jurkat-TCR D activation by peptide, as measure by CD69 expression, was dose responsive, with ECso of 45.54 ng / mL (FIG. 1). The results demonstrated that TCR D was able to induce TCR signaling and has the potential to induce CLAVEEVSL-specific cytotoxic signaling if engineered into primary cells.Example 9. TCR-mediated toxicity against target cells.

[0280] Anti -CLAVEEVSL TCRs were evaluated for cytotoxicity against HLA*A02:01 AML cell lines (0CI-AML3) which express a NPMlc mutation. In brief, primary CD8+T cells from HLA*A02:01 negative healthy donors were engineered to knock-out the endogenous TCR a and P constant chain domains. Subsequently, the CD8+cells were transduced with TCRs. 0CI-AML2 cells (HLA*A02:01+) were peptide pulsed with the target peptide and subsequently co-cultured with the engineered primary CD8+T cells to evaluate TCRcytotoxicity.

[0281] Frozen PBMCs were stimulated with pyrogenic CD3 antibody prior to purification of activated CD8 T cells. Subsequently the cells were endogenously TCR deleted by CRISPR and lentivirally transduced with TCR D, TCR E, or TCR F. Primary cells were expanded for 10 days in CTS media with serum replacement optimizer supplemented with 50 U of IL-2 and 10 ng / mL of IL-7. After 10 days transduced T cells were purified using CD34 (transduction marker) bead selection. Engineering of the CD8+TCR-T was confirmed by FACS analysis using CD3, CD8 (primary T cell specific marker), CD34 & NPM1 specific tetramer and live dead staining.

[0282] For cytotoxicity assays, 0CI-AML2 cells were peptide pulsed with titrated concentrations of CLAVEEVSL (SEQ ID NO: 5) peptide for 1 h prior to co-culture.

[0283] Engineered CD8+TCR-T cells were co-cultured with control or peptide pulsed OCL AML2 cells at an E:T ratio of 1 : 1 in 96 well plates or with 0CI-AML3 cells or 0CI-AML2 cells (negative control cells) at varying E:T ratios in 96 well plates. For TCR-T negative control conditions, HPV specific TCR-T cells were engineered as described above and co-cultured as described above. No cytotoxicity was observed against cells not expressing or not pulsed with the target peptide, (data not shown). After 16-18 h, the co-cultured cells were stained with fluorescently tagged antibodies or cell dyes for CD8 (primary T cell specific marker), VPD450 (to identify 0CI-AML2 cells or 0CI-AML3 cells) and live / dead staining. Data are presented as the percent of pulsed (with target peptide) vs. non-pulsed (without target peptide) OCL AML2 cells (FIG. 2) or percent of OCI-AML3 (expressing the target peptide) vs. non-pulsed (FIG. 3). The results, shown for TCR D, indicates the TCRs were effective in killing cells pulsed with or naturally expressing the target peptide.

[0284] Co-culture assays were performed as described above with multiple TCRs and donors. The previously published Leiden (US11352389) TCR was included as a control. Data are presented as the percent of peptide pulsed OCI-AML2 cells (sensitive to immunogenic killing) vs OCI-AML2 cells without target peptide. ECso for peptide induced TCR cytotoxicity was determined based on the target cell numbers of peptide pulsed OCI-AML2 cells.Example 10. E:T EC so determination

[0285] Co-culture assays were performed as described above using TCRD, TCRE, and TCR F engineered T cells that were harvested from three healthy donors and three different donor cells known to express NPMlc. The previously published Leiden TCR (US11352389) was included as a control. For cytotoxicity, E:T ratio determined ECsos for engineered T cell wasdetermined based on OCI-AML3 target cell numbers. The data in Table 7 demonstrate that engineered T cells expressing TCR D, TCR E, or TCR F have potent cytotoxicity activity to cell lines expressing the NPMlc antigen CLAVEEVSL (SEQ ID NO: 5), each having an EC50 E:T ratio less than 1. Furthermore, the observed cytotoxicity was not significantly different published engineered T cells. The demonstrated killing of 0CI-AML3 cells by TCR D, TCR E, and TCR F engineered T cells is indicative of mutation specific cytotoxicity for cells expressing NPMlc mutations which are presented in patient AML blast cells.Table 7. Killing of donor target cells with engineered T cells expressing TCR D, TCR E, or TCR F.Example 11. Tetramer based sorting of CLAVEEVSL reacting CD8+ T cell clones.

[0286] T cell isolation was performed by stimulating T cells with activated dendritic cells. Dendritic cells were generated by stimulating purified CD14 monocytes, derived from PBMCs, with 10 ng / mL LPS, IL-4, INF-y and GM-CSF for 4 days. The stimulated dendritic cells were pulsed with 10 ng / mL CLAVEEVL (SEQ ID NO:5) peptide on day 5. From day 1 to day 4, isolated CD8 T cells from the same donor were maintained in XVIVO-15 media with 5% human serum. CD8+cells were co-cultured with pulsed dendritic cells. Cells were maintained in XVIVO-15 media with the addition of 30 ng / mL IL-21 on day 5 and subsequently supplemented with 5 ng / mL IL-7 and IL-15 on days 8, 10 and 12. On day 16, CD8+T cells were stained with two 1 :40 CLAVEEVSL tetramers stained for allophycocyanin (APC) and R- phycoerythrin(PE) for 1 h at room temperature and then FACS sorted for tetramer positive T cells. The positive T cells were cloned and sequenced prior to further evaluation of the TCRs identified by tetramer double positive sorting.

[0287] Tetramer based sorting of CD8+T cell clones was also conducted for the AIQDCLAV (SEQ ID NO: 1) peptide as described above for the CLAVEEVSL (SEQ ID NO:5) peptide (data not shown).

[0288] Stable transduced Jurkat cells were stained with CD3, CD34 and CLAVEEVSL orC*LAVEEVSL tetramer (1 :40) to validate TCR / peptide specificity.

[0289] We were able to isolate unique TCRs to the NPMlc CLAVEEVSL and AIQDCLAV neoantigen peptide tetramers as demonstrated by FACS plots. Additionally, we were able to demonstrate that the isolated anti-CLAVEEVLS TCRs were able to bind both un-cysteinylated CLAVEEVSL and cysteinylated C*LAVEEVSL tetramers. Binding to the C*LAVEEVSL (SEQ ID NO:5) has been reported to be important for killing of mutNPMlc target cells (van der Lee DI et al., “Mutated nucleophosmin 1 as immunotherapy target in acute myeloid leukemia” J Clin Invest 2019 129(2):774-785). Engineered cells expressing TCR D exhibited, in separate experiments, 34.40% and 39.8% CLAVEEVSL tetramer binding (by FACS analysis) and 83.85% and 83.24% C*LAVEEVSL tetramer binding.

[0290] Additional activation, tetramer binding, and EC50 data are shown in Tables 8 and 9.Table 8. Binding of TCRs to target cells.*Primary CD8+ T cells co-cultured with 0CI-AML3**Primary CD8+ T cells co-cultured with 0CI-AML2 and CLAY peptide pulsedTable 9. Binding of TCRs to target cells.Example 12. Modified TCRs.

[0291] TCR D was engineered (modified) to introduce an additional disulfide bond to reduce the likelihood that its a or P chains would form chimeric TCRs with the endogenous primary CD8 TCR, thus reducing potential toxicity due to chimeric TCRs that do not target the NPMlc neoepitope. The TCR D was modified to introduce cysteines at positions 186 of the a chain (T186C substitution, position relative to SEQ ID NO:71) and position 191 of the P chain (S191C, position relative to SEQ ID NO:79), see Table 10.

[0292] Jurkat cells were transduced with the unmodified TCR D or the disulfide modified TCR D, as previously described. The disulfide modified TCR D bound exhibited increase binding to CLAVEEVSL (95.48%) tetramer relative to TCR D (88.45%), as determined by FACS analysis. Thus, the disulfide bond improves potency, possibly by increasing the proportion of anti-NPMlc TCR a / p pairing in engineered T cells. The same improvement in potency is expected to be seen if similar cysteine substitutions are made in the other described TCRs.

[0293] In addition, TCR D was also engineered with a and P constant regions being replaced with Ig domains of the y and 5 constant regions. Jurkat cells were transduced with the unmodified TCR D or the domain swapped TCR D as previously described. No tetramer binding was observed with the negative control domain swapped TCR Ds.Table 10. TCR modifications.Example 13. Engineered T cells are superior to CAR T cells.

[0294] An anti-NPMlc chimeric T cell receptor was generated comprising an anti- AIQDCLAV scFv as described (Xie G et al. “CAR-T cells targeting a nucleophosmin neoepitope exhibit potent specific activity in mouse models of acute myeloid leukaemia” Nat Biomed Eng. 2021), with modifications. The modifications were the use of a P2A element instead of a T2A elements and an RQR8 tag instead of an EGFR tag. It is known in the art thatthese modifications do not affect the targeting of the 5' CAR-T construct.

[0295] Engineered T cells and CAR-T cells were manufactured by isolating CD8+T cells, activating with TransACT and expanded using G-REX. Transduction, CRISPR editing, and cytokine supplementation was performed as previously described.

[0296] Reporter assays were performed as described above to evaluate anti-AIQDCLAV CAR-T cells and engineered TCR B T cells. Target cells were pulsed with AIQDCLAV (SEQ ID NO: 1) target peptide as described above for CLAVEEVSL (SEQ ID NO:5) peptide. Additionally, engineered TCR D T cells were evaluated in parallel against CLAVEEVSL (SEQ ID NO:5) target peptide pulsed cells. In pulse peptide reporter assays utilizing K562 cells as antigen presenting cells, HLA*A02:01 were used and K562 HLA*A24:02 cells were used as negative control.

[0297] Engineered Jurkat cells transduced with chimeric T cell receptor, TCRB, or TCR D were analyzed for reactivity against target cells A*02:01 K562s or A*24:02 K562s exogenously loaded with titrated concentrations of target peptide. The cells were co-cultured for 16-18 h at an E:T ratio of 1 : 1. The value of activation of the Jurkat cells was evaluated as the percentage of CD69+cells by flow cytometry (FIG. 4). Engineered TCR D T cells demonstrated robust TCR signal induction. Engineered anti-AIQDCLAV (TCR B) T cells also induced robust TCR signaling as determined by CD69 expression. Both the TCR B and TCR D engineered T cells exhibited substantially greater signaling than the CAR-T cells. As expected, no signaling was observed with K562 HLA*A24:02 cells.

[0298] TCR B and TCR D exhibit superior CD69 reporter signaling relative to published (Xie et al. 2021) anti-AIQDCLAV CAR-T cells in Jurkat CD 8 positive cells co-cultured with NPM1 pulsed K562 cells expressing HLA-A*02:01.Example 14. Comparison of CAR-T vs TCR-T in cytotoxicity assays.

[0299] Primary CD8+cells, from healthy donor, gene edited and transduced with TCR D (Native) or disulfide added TCR D (DB), or anti-AIQDCLAV CAR-T cells were tested for recognition of HLA-A*02:01 immunogenic AML cell lines with NPMlc (OCI-AML3) as previously described. The cells were co-cultured for 16-18 h at different E:T ratios. Percent killing was evaluated by flowcytometry and the EC so were generated in GraphPad Prism (FIG. 5).

[0300] Both the TCR D and disulfide modified TCR D engineered T cells exhibited substantially greater killing and lower ECso relative to CAR-T cells. Because the endogenous TCR was deleted in the engineered T cells, no difference in killing was observed between theTCR D and disulfide added TCR D engineered T cells. TCR knockout (KO) T cells were used as a negative control.Example 15. TCR D targets neoantigen (CLAVEEVSL) with and without post translational modifications.

[0301] Killing assay were performed as previously described, with minor modifications. Cytotoxicity assays were performed with OCI-AML2 cells pulsed with either CLAVEEVSL (SEQ ID NO:5) peptide lacking post-translation modifications or with cysteinylation of the first cysteine in the target peptide (indicated as C*LAVEEVSL). Pulsed peptide assays were performed at an E:T ratio of 0.5: 1 and 1 : 1 with 10 ng / mL of peptide. Wild-type NPM1 was used as a negative peptide control and an anti-HPV TCR engineered T cell was used as negative engineered T cell control.

[0302] It has been shown (van der Lee et al. 2019) that cytotoxicity towards C*LAVEEVSL is important for T cell therapy against NPMlc+cancers. Previously published anti- CLAVEEVSL TCRs show showed differential activation, as measured by INFy, when binding to CLAVEEVSL or C*LAVEEVSL. Clonal reactivity was more potent against the uncysteinylated target peptide. In contrast, TCR D engineered T cells demonstrated high cytotoxicity to both the cysteinylated and uncysteinylated target peptide (FIGs. 7-8), suggesting potential cytotoxicity advantages over previously published anti-CLAVEEVSL TCRs (US 11352389).Example 16. Identification of amino acids of CLAVEEVSL (SEQ ID NO: 5) recognized by the anti-CLAVEEVSL TCRs.

[0303] Alanine scanning of the CLAVEEVSL (SEQ ID NO: 5) peptides was performed. The peptides used are as described in Table 11.

[0304] Reporter assays were performed as previously described with minor modifications. In brief target K562 cells were pulsed with the listed peptides at various peptide concentrations for 1 h. CD8+Jurkat cells were engineered to express TCR D and TCR F. Reporter assays were performed using E:T ratio of 1 : 1. Control conditions included the wild type negative control peptide or the immunogenic positive control peptide (CLAVEEVSL, SEQ ID NO: 5).Table 11. Alanine scan CLAVEEVSL (SEQ ID NO:5) peptides

[0305] Residues of CLAVEEVSL (SEQ ID NO: 5) important in recognition by the anti- CLAVEEVLS TCRs was assessed by the reduction or loss of CD69 signaling when an amino acid was replaced with alanine. Amino acids at the 2nd, 5th, 6th, and 7thpositions (relative to SEQ ID NO:5) were found to be important for recognition by both TCRs (FIG. 7 and 8) Surprisingly, residue 4 was only important for recognition by TCR D and residue 9 was only important for recognition by TCR F.

[0306] The present invention is not intended to be limited in scope to the particular disclosed embodiments, which are provided, for example, to illustrate various aspects of the invention. Various modifications to the compositions and methods described will become apparent from the description and teachings herein. Such variations may be practiced without departing from the true scope and spirit of the disclosure and are intended to fall within the scope of the present disclosure.Table 12. Sequences

Claims

Claims:

1. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising: an a chain comprising a variable a (Va) region and a P chain comprising a variable P (VP) region; or a y chain comprising a variable y (Vy) region and a 5 chain comprising a variable 5 (V5) region; wherein:(a) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:67, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:75;(b) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:85, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:93;(c) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 103, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 111;(d) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:31, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:39;(e) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:49, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:57;(f) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 121, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 129;(g) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 139, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 147;(h) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 157, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 165;(i) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 175, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO: 183;(j) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO: 193, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:201;(k) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:211, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:219;(l) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:229, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:237;(m) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:247, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:255;(n) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:265, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:273;(o) the Va or Vy region comprises a CDR-3 comprising SEQ ID NO:283, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:291; or(p) the Va or Vy region comprises a complementarity determining region 3 (CDR- 3) comprising SEQ ID NO: 13, and the VP or V5 region comprises a CDR-3 comprising SEQ ID NO:21.

2. The TCR or antigen-binding fragment thereof of claim 1, wherein:(a) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:65, and a CDR-2 comprising SEQ ID NO:66, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:73, and a CDR-2 comprising SEQ ID NO:74;(b) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:83, and a CDR-2 comprising SEQ ID NO:84, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:91, and a CDR-2 comprising SEQ ID NO:92;(c) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 101, and a CDR-2 comprising SEQ ID NO: 102, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 109, and a CDR-2 comprising SEQ ID NO: 110;(d) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:29, and a CDR-2 comprising SEQ ID NO:30, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:37, and a CDR-2 comprising SEQ ID NO:38;(e) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:47, and a CDR-2 comprising SEQ ID NO:48, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:55, and a CDR-2 comprising SEQ ID NO:56;(f) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 119, and a CDR-2 comprising SEQ ID NO: 120, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 127, and a CDR-2 comprising SEQ ID NO: 128;(g) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 137, and a CDR-2 comprising SEQ ID NO: 138, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 145, and a CDR-2 comprising SEQ ID NO: 146;(h) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 155, and a CDR-2 comprising SEQ ID NO: 156, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 163, and a CDR-2 comprising SEQ ID NO: 164;(i) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 173, and a CDR-2 comprising SEQ ID NO: 174, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 181, and a CDR-2 comprising SEQ ID NO: 182;(j) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 191, and a CDR-2 comprising SEQ ID NO: 192, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 199, and a CDR-2 comprising SEQ ID NO:200;(k) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:209, and a CDR-2 comprising SEQ ID NO:210, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:217, and a CDR-2 comprising SEQ ID NO:218;(l) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:227, and a CDR-2 comprising SEQ ID NO:228, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:235, and a CDR-2 comprising SEQ ID NO:236;(m) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:245, and a CDR-2 comprising SEQ ID NO:246, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:253, and a CDR-2 comprising SEQ ID NO:254;(n) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:263, and a CDR-2 comprising SEQ ID NO:264, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:271, and a CDR-2 comprising SEQ ID NO:272;(o) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:281, and a CDR-2 comprising SEQ ID NO:282, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:289, and a CDR-2 comprising SEQ ID NO:290; or(p) the Va or Vy region comprises a complementarity determining region 1 (CDR- 1) comprising SEQ ID NO: 11, and a complementarity determining region 2 (CDR-2) comprising SEQ ID NO: 12, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 19, and a CDR-2 comprising SEQ ID NO:20.

3. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising: an a chain comprising a variable a (Va) region and a P chain comprising a variable P (VP) region; or a y chain comprising a variable y (Vy) region and a 5 chain comprising a variable 5 (V5) region; wherein:(a) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:65, a CDR-2 comprising SEQ ID NO:66, and a CDR-3 comprising SEQ ID NO:67, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:73, a CDR-2 comprising SEQ ID NO:74, and a CDR-3 comprising SEQ ID NO:75;(b) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:83, a CDR-2 comprising SEQ ID NO:84, and a CDR-3 comprising SEQ ID NO:85, and the VP or V5 regioncomprises a CDR-1 comprising SEQ ID NO:91, a CDR-2 comprising SEQ ID NO:92, and a CDR-3 comprising SEQ ID NO:93;(c) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 101, a CDR-2 comprising SEQ ID NO: 102, and a CDR-3 comprising SEQ ID NO: 103, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 109, a CDR-2 comprising SEQ ID NO: 110, and a CDR-3 comprising SEQ ID NO: 111;(d) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:29, a CDR-2 comprising SEQ ID NO:30, and a CDR-3 comprising SEQ ID NO:31, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:37, a CDR-2 comprising SEQ ID NO:38, and a CDR-3 comprising SEQ ID NO:38;(e) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:47, a CDR-2 comprising SEQ ID NO:48, and a CDR-3 comprising SEQ ID NO:49, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:55, a CDR-2 comprising SEQ ID NO:56, and a CDR-3 comprising SEQ ID NO:57;(f) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 119, a CDR-2 comprising SEQ ID NO: 120, and a CDR-3 comprising SEQ ID NO: 121, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 127, a CDR-2 comprising SEQ ID NO: 128, and a CDR-3 comprising SEQ ID NO: 129;(g) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 137, a CDR- 2 comprising SEQ ID NO: 138, and a CDR-3 comprising SEQ ID NO: 139, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 145, a CDR-2 comprising SEQ ID NO: 146, and a CDR-3 comprising SEQ ID NO: 147;(h) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 155, a CDR- 2 comprising SEQ ID NO: 156, and a CDR-3 comprising SEQ ID NO: 157, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 163, a CDR-2 comprising SEQ ID NO: 164, and a CDR-3 comprising SEQ ID NO: 165;(i) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 173, a CDR-2 comprising SEQ ID NO: 174, and a CDR-3 comprising SEQ ID NO: 175, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 181, a CDR-2 comprising SEQ ID NO: 182, and a CDR-3 comprising SEQ ID NO: 183;(j) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 191, a CDR-2 comprising SEQ ID NO: 192, and a CDR-3 comprising SEQ ID NO: 193, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 199, a CDR-2 comprising SEQ IDNO:200, and a CDR-3 comprising SEQ ID NO:201;(k) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:209, a CDR- 2 comprising SEQ ID NO:210, and a CDR-3 comprising SEQ ID NO:211, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO :217, a CDR-2 comprising SEQ IDNO:218, and a CDR-3 comprising SEQ ID NO:219;(l) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:227, a CDR-2 comprising SEQ ID NO:228, and a CDR-3 comprising SEQ ID NO:229, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:235, a CDR-2 comprising SEQ IDNO:236, and a CDR-3 comprising SEQ ID NO:237;(m) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:245, a CDR- 2 comprising SEQ ID NO:246, and a CDR-3 comprising SEQ ID NO:247, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:253, a CDR-2 comprising SEQ IDNO:254, and a CDR-3 comprising SEQ ID NO:255;(n) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:263, a CDR- 2 comprising SEQ ID NO:264, and a CDR-3 comprising SEQ ID NO:265, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:271, a CDR-2 comprising SEQ IDNO:272, and a CDR-3 comprising SEQ ID NO:273;(o) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO:281, a CDR- 2 comprising SEQ ID NO:282, and a CDR-3 comprising SEQ ID NO:283, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO:289, a CDR-2 comprising SEQ IDNO:290, and a CDR-3 comprising SEQ ID NO:291; or(p) the Va or Vy region comprises a CDR-1 comprising SEQ ID NO: 11, a CDR-2 comprising SEQ ID NO: 12, and a CDR-3 comprising SEQ ID NO: 13, and the VP or V5 region comprises a CDR-1 comprising SEQ ID NO: 19, a CDR-2 comprising SEQ ID NO:20, and a CDR-3 comprising SEQ ID NO:21.

4. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising: an a chain comprising a variable a (Va) region and a P chain comprising a variable P (VP) region; or a y chain comprising a variable y (Vy) region and a 5 chain comprising a variable 5 (V5) region; wherein:(a) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:68, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:76;(b) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy regionsequence of SEQ ID NO: 86, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 94;(c) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 104, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 112;(d) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR- 3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:40;(e) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 50, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:58;(f) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 122, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 130;(g) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 140, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 148;(h) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 158, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 166;(i) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 176, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 184;(j) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 194, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:202;(k) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:212, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:220;(l) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:230, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:238;(m) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:248, and the VP or V5 region comprises a CDR-3 as containedwithin the VP or V5 region sequence of SEQ ID NO:256;(n) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:266, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:274;(o) the Va or Vy region comprises a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:284, and the VP or V5 region comprises a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:292;(p) the Va or Vy region comprises a CDR-3 comprising a CDR-3 contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR- 3 comprising a CDR-3 contained within the VP or V5 region sequence of SEQ ID NO:22.

5. The TCR or antigen-binding fragment thereof of claim 4, wherein:(a) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:68, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:76;(b) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:86, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:94;(c) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO: 104, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO: 112;(d) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:40;(e) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:50, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:58;(f) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO: 122, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO: 130;(g) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO: 140, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO: 148;(h) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO: 158, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO: 166;(i) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO: 176, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO: 184;(j) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO: 194, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:202;(k) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:212, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:220;(l) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:230, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:238;(m) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:248, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:256;(n) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:266, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:274;(o) the Va or Vy region comprises a CDR-1 and a CDR-2 as contained within the Va or Vy region sequence of SEQ ID NO:284, and the VP or V5 region comprises a CDR-1 and a CDR-2 as contained within the VP or V5 region sequence of SEQ ID NO:292; or(p) the Va or Vy region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR-1 and a CDR-2 comprising a CDR-1 and a CDR-2, respectively, contained within the VP or V5 region sequence of SEQ ID NO:22.

6. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising: an a chain comprising a variable a (Va) region and a P chain comprising a variable P (VP) region; or a y chain comprising a variable y (Vy) region and a 5 chain comprising a variable 5 (V5) region; wherein:(a) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:68, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:76;(b) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:86, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 94;(c) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 104, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 112;(d) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO:32, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:40;(e) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:50, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:58;(f) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 122, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 130;(g) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 140, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 148;(h) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 158, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 166;(i) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as containedwithin the Va or Vy region sequence of SEQ ID NO: 176, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO: 184;(j) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO: 194, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:202;(k) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:212, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:220;(l) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:230, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:238;(m) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:248, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:256;(n) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:266, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:274;(o) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the Va or Vy region sequence of SEQ ID NO:284, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 as contained within the VP or V5 region sequence of SEQ ID NO:292;(p) the Va or Vy region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the Va or Vy region sequence of SEQ ID NO: 14, and the VP or V5 region comprises a CDR-1, a CDR-2, and a CDR-3 comprising a CDR-1, a CDR-2, and a CDR-3, respectively, contained within the VP or V5 region sequence of SEQ ID NO:22.

7. A T cell receptor (TCR) or antigen-binding fragment thereof, comprising:an a chain comprising a variable a (Va) region and a P chain comprising a variable P (VP) region; or a y chain comprising a variable y (Vy) region and a 5 chain comprising a variable 5 (V5) region; wherein:(a) the Va or Vy region comprises SEQ ID NO:68 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:76 or a sequence that has at least 90% sequence identity thereto;(b) the Va or Vy region comprises SEQ ID NO:86 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:94 or a sequence that has at least 90% sequence identity thereto;(c) the Va or Vy region comprises SEQ ID NO: 104 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 112 or a sequence that has at least 90% sequence identity thereto;(d) the Va or Vy region comprises SEQ ID NO:32 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:40 or a sequence that has at least 90% sequence identity thereto;(e) the Va or Vy region comprises SEQ ID NO:50 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:58 or a sequence that has at least 90% sequence identity thereto;(f) the Va or Vy region comprises SEQ ID NO: 122 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 130 or a sequence that has at least 90% sequence identity thereto;(g) the Va or Vy region comprises SEQ ID NO: 140 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 148 or a sequence that has at least 90% sequence identity thereto;(h) the Va or Vy region comprises SEQ ID NO: 158 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 166 or a sequence that has at least 90% sequence identity thereto;(i) the Va or Vy region comprises SEQ ID NO: 176 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO: 184 or a sequence that has at least 90% sequence identity thereto;(j) the Va or Vy region comprises SEQ ID NO: 194 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:202 or a sequence that has at least 90% sequence identity thereto;(k) the Va or Vy region comprises SEQ ID NO:212 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:220 or a sequence that has at least 90% sequence identity thereto;(l) the Va or Vy region comprises SEQ ID NO:230 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:238 or a sequence that has at least 90% sequence identity thereto;(m) the Va or Vy region comprises SEQ ID NO:248 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:256 or a sequence that has at least 90% sequence identity thereto;(n) the Va or Vy region comprises SEQ ID NO:266 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:274 or a sequence that has at least 90% sequence identity thereto;(o) the Va or Vy region comprises SEQ ID NO:284 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:292 or a sequence that has at least 90% sequence identity thereto; or(p) the Va or Vy region comprises SEQ ID NO: 14 or a sequence that has at least 90% sequence identity thereto, and the VP or V5 region comprises SEQ ID NO:22 or a sequence that has at least 90% sequence identity thereto.

8. The TCR or antigen-binding fragment thereof of any one of claims 1-7, wherein:(a) the Va or Vy region comprises SEQ ID NO:68, and the VP or V5 region comprises SEQ ID NO:76;(b) the Va or Vy region comprises SEQ ID NO:86, and the VP or V5 region comprises SEQ ID NO:94;(c) the Va or Vy region comprises SEQ ID NO: 104, and the VP or V5 region comprises SEQ ID NO: 112;(d) the Va or Vy region comprises SEQ ID NO:32, and the VP or V5 region comprises SEQ ID NO:40.(e) the Va or Vy region comprises SEQ ID NO:50, and the VP or V5 region comprises SEQ ID NO:58;(f) the Va or Vy region comprises SEQ ID NO: 122, and the VP or V5 region comprises SEQ ID NO: 130;(g) the Va or Vy region comprises SEQ ID NO: 140, and the VP or V5 regioncomprises SEQ ID NO: 148;(h) the Va or Vy region comprises SEQ ID NO: 158, and the VP or V5 region comprises SEQ ID NO: 166;(i) the Va or Vy region comprises SEQ ID NO: 176, and the VP or V5 region comprises SEQ ID NO: 184;(j) the Va or Vy region comprises SEQ ID NO: 194, and the VP or V5 region comprises SEQ ID NO:202;(k) the Va or Vy region comprises SEQ ID NO:212, and the VP or V5 region comprises SEQ ID NO:220;(l) the Va or Vy region comprises SEQ ID NO:230, and the VP or V5 region comprises SEQ ID NO:238;(m) the Va or Vy region comprises SEQ ID NO:248, and the VP or V5 region comprises SEQ ID NO:256;(n) the Va or Vy region comprises SEQ ID NO:266, and the VP or V5 region comprises SEQ ID NO:274;(o) the Va or Vy region comprises SEQ ID NO:284, and the VP or V5 region comprises SEQ ID NO:292; or(p) the Va or Vy region comprises SEQ ID NO: 14, and the VP or V5 region comprises SEQ ID NO:22.

9. The TCR or antigen-binding fragment thereof of any one of claims 1-8, wherein: the a chain further comprises an a constant (Ca) region and the P chain further comprises a P constant (CP) region; or the y chain further comprises an y constant (Cy) region and the 5 chain further comprises a 5 constant (C5) region.

10. The TCR or antigen-binding fragment thereof of claim 9, wherein the Ca comprises SEQ ID NO:3 and the CP comprises SEQ ID NO:7.

11. The TCR or antigen-binding fragment thereof of any one of claims 1-10, wherein:(a) the alpha or gamma chain comprises SEQ ID NO:71, and the beta or delta chain comprises SEQ ID NO:79;(b) the alpha or gamma chain comprises SEQ ID NO:89, and the beta or delta chaincomprises SEQ ID NO:97;(c) the alpha or gamma chain comprises SEQ ID NO: 107, and the beta or delta chain comprises SEQ ID NO: 115;(d) the a or y chain comprises SEQ ID NO:35, and the P or 5 chain comprises SEQ ID NO:43;(e) the alpha or gamma chain comprises SEQ ID NO:53, and the beta or delta chain comprises SEQ ID NO:61;(f) the alpha or gamma chain comprises SEQ ID NO: 125, and the beta or delta chain comprises SEQ ID NO: 133;(g) the alpha or gamma chain comprises SEQ ID NO: 143, and the beta or delta chain comprises SEQ ID NO: 151;(h) the alpha or gamma chain comprises SEQ ID NO: 161, and the beta or delta chain comprises SEQ ID NO: 169;(i) the alpha or gamma chain comprises SEQ ID NO: 179, and the beta or delta chain comprises SEQ ID NO: 187;(j) the alpha or gamma chain comprises SEQ ID NO: 197, and the beta or delta chain comprises SEQ ID NO:205;(k) the alpha or gamma chain comprises SEQ ID NO:215, and the beta or delta chain comprises SEQ ID NO:223;(l) the alpha or gamma chain comprises SEQ ID NO:233, and the beta or delta chain comprises SEQ ID NO:241;(m) the alpha or gamma chain comprises SEQ ID NO:251, and the beta or delta chain comprises SEQ ID NO:259;(n) the alpha or gamma chain comprises SEQ ID NO:269, and the beta or delta chain comprises SEQ ID NO:277;(o) the alpha or gamma chain comprises SEQ ID NO:287, and the beta or delta chain comprises SEQ ID NO:295; or(p) the a or y chain comprises SEQ ID NO: 17, and the P or 5 chain comprises SEQ ID NO:25.

12. The TCR or antigen-binding fragment thereof of any one of claims 1-11, wherein the TCR or antigen-binding fragment thereof recognizes a NPMlc neoantigen in the context of an MHC molecule.

13. The TCR or antigen-binding fragment thereof of claim 12, wherein the MHC molecule is a human leukocyte antigens (HLA)-A molecule.

14. The TCR or antigen-binding fragment thereof of claim 13, wherein the HLA- A molecule is of serotype HLA-A*02:01, HLA-A*02:06, or HLA-A*02:03.

15. The TCR or antigen-binding fragment thereof of any one of claims 12-14, wherein the NPMlc neoantigen comprises CLAVEEVSL (SEQ ID NO:5) or AIQDLCLAV (SEQ ID NO:1) or.

16. The TCR or antigen-binding fragment thereof of 15, wherein the TCR or antigen-binding fragment thereof(a) recognizes CLAVEEVSL (SEQ ID NO: 5). with higher affinity than AIQDLWQWRKSL (SEQ ID NO:2); or(b) recognizes AIQDLCLAV (SEQ ID NO: 1). with higher affinity than AIQDLWQWRKSL (SEQ ID NO:2).

17. Nucleic acid sequences encoding the TCR or antigen-binding fragment thereof of any one of claims 1-16, or an a chain, a P chain, a y chain, or a 5 chain thereof.

18. The nucleic acid sequences of claim 17, wherein the nucleic acid sequences comprise a first nucleotide sequence encoding the Va region and a second nucleotide sequence encoding the VP region; or a first nucleotide sequence encoding the Vy region and a second nucleotide sequence encoding the V5 region; wherein:(a) the first nucleotide sequence comprises SEQ ID NO:69 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:77 or a sequence that has at least 90% sequence identity thereto;(b) the first nucleotide sequence comprises SEQ ID NO:87 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 95 or a sequence that has at least 90% sequence identity thereto;(c) the first nucleotide sequence comprises SEQ ID NO: 105 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 113 or a sequence that has at least 90% sequence identity thereto;(d) the first nucleotide sequence comprises SEQ ID NO:33 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:41 or a sequence that has at least 90% sequence identity thereto;(e) the first nucleotide sequence comprises SEQ ID NO:51 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:59 or a sequence that has at least 90% sequence identity thereto;(f) the first nucleotide sequence comprises SEQ ID NO: 123 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 131 or a sequence that has at least 90% sequence identity thereto;(g) the first nucleotide sequence comprises SEQ ID NO: 141 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 149 or a sequence that has at least 90% sequence identity thereto;(h) the first nucleotide sequence comprises SEQ ID NO: 159 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 167 or a sequence that has at least 90% sequence identity thereto;(i) the first nucleotide sequence comprises SEQ ID NO: 177 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 185 or a sequence that has at least 90% sequence identity thereto;(j) the first nucleotide sequence comprises SEQ ID NO: 195 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:203 or a sequence that has at least 90% sequence identity thereto;(k) the first nucleotide sequence comprises SEQ ID NO:213 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:221 or a sequence that has at least 90% sequence identity thereto;(l) the first nucleotide sequence comprises SEQ ID NO:231 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:239 or a sequence that has at least 90% sequence identity thereto;(m) the first nucleotide sequence comprises SEQ ID NO:249 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:257 or a sequence that has at least 90% sequence identity thereto;(n) the first nucleotide sequence comprises SEQ ID NO:267 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:275 or a sequence that has at least 90% sequence identity thereto;(o) the first nucleotide sequence comprises SEQ ID NO:285 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:293 or a sequence that has at least 90% sequence identity thereto;(p) the first nucleotide sequence comprises SEQ ID NO: 15 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:23 or a sequence that has at least 90% sequence identity thereto;(aa) the first nucleotide sequence comprises SEQ ID NO:70 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:78 or a sequence that has at least 90% sequence identity thereto;(bb) the first nucleotide sequence comprises SEQ ID NO:88 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 96 or a sequence that has at least 90% sequence identity thereto;(cc) the first nucleotide sequence comprises SEQ ID NO: 106 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 114 or a sequence that has at least 90% sequence identity thereto;(dd) the first nucleotide sequence comprises SEQ ID NO:34 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:42 or a sequence that has at least 90% sequence identity thereto;(ee) the first nucleotide sequence comprises SEQ ID NO:52 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 60 or a sequence that has at least 90% sequence identity thereto;(ff) the first nucleotide sequence comprises SEQ ID NO: 124 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 132 or a sequence that has at least 90% sequence identity thereto;(gg) the first nucleotide sequence comprises SEQ ID NO: 142 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 150 or a sequence that has at least 90% sequence identity thereto;(hh) the first nucleotide sequence comprises SEQ ID NO: 160 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 168 or a sequence that has at least 90% sequence identity thereto;(ii) the first nucleotide sequence comprises SEQ ID NO: 178 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 186 or a sequence that has at least 90% sequence identity thereto;(jj) the first nucleotide sequence comprises SEQ ID NO: 196 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:204 or a sequence that has at least 90% sequence identity thereto;(kk) the first nucleotide sequence comprises SEQ ID NO:214 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:222 or a sequence that has at least 90% sequence identity thereto;(11) the first nucleotide sequence comprises SEQ ID NO:232 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:240 or a sequence that has at least 90% sequence identity thereto;(mm) the first nucleotide sequence comprises SEQ ID NO:250 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:258 or a sequence that has at least 90% sequence identity thereto;(nn) the first nucleotide sequence comprises SEQ ID NO:268 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:276 or a sequence that has at least 90% sequence identity thereto;(oo) the first nucleotide sequence comprises SEQ ID NO:286 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:294 or a sequence that has at least 90% sequence identity thereto; or(pp) the first nucleotide sequence comprises SEQ ID NO: 16 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:24 or a sequence that has at least 90% sequence identity thereto.

19. The nucleic acid sequences of claim 17 or 18, wherein:(a) the first nucleotide sequence comprises SEQ ID NO:72 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 80 or a sequence that has at least 90% sequence identity thereto;(b) the first nucleotide sequence comprises SEQ ID NO:90 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 98 or a sequence that has at least 90% sequence identity thereto;(c) the first nucleotide sequence comprises SEQ ID NO: 108 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 116 or a sequence that has at least 90% sequence identity thereto;(d) the first nucleotide sequence comprises SEQ ID NO:36 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:44 or a sequence that has at least 90% sequence identity thereto;(e) the first nucleotide sequence comprises SEQ ID NO:54 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ IDNO: 62 or a sequence that has at least 90% sequence identity thereto;(f) the first nucleotide sequence comprises SEQ ID NO: 126 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 134 or a sequence that has at least 90% sequence identity thereto;(g) the first nucleotide sequence comprises SEQ ID NO: 144 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 152 or a sequence that has at least 90% sequence identity thereto;(h) the first nucleotide sequence comprises SEQ ID NO: 162 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 170 or a sequence that has at least 90% sequence identity thereto;(i) the first nucleotide sequence comprises SEQ ID NO: 180 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO: 188 or a sequence that has at least 90% sequence identity thereto;(j) the first nucleotide sequence comprises SEQ ID NO: 198 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:206 or a sequence that has at least 90% sequence identity thereto;(k) the first nucleotide sequence comprises SEQ ID NO:216 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:224 or a sequence that has at least 90% sequence identity thereto;(l) the first nucleotide sequence comprises SEQ ID NO:234 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:242 or a sequence that has at least 90% sequence identity thereto;(m) the first nucleotide sequence comprises SEQ ID NO:252 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:260 or a sequence that has at least 90% sequence identity thereto;(n) the first nucleotide sequence comprises SEQ ID NO:270 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:278 or a sequence that has at least 90% sequence identity thereto;(o) the first nucleotide sequence comprises SEQ ID NO:288 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:296 or a sequence that has at least 90% sequence identity thereto; or(p) the first nucleotide sequence comprises SEQ ID NO: 18 or a sequence that has at least 90% sequence identity thereto, and the second nucleotide sequence comprises SEQ ID NO:26 or a sequence that has at least 90% sequence identity thereto.

20. The nucleic acid sequences of any one of claims 17-19 wherein the first and second nucleotide are present on a single expression vector and expressed from a single promoter.

21. The nucleic acid sequences of any one of claims 17-20, wherein the first and second nucleotide sequences are separated by a peptide sequence that causes ribosome skipping.

22. The nucleic acid sequences of claim 21, wherein the peptide that causes ribosome skipping is a P2A peptide.

23. The nucleic acid sequences of claim 22, wherein the P2A peptide comprises SEQ ID NO:301.

24. The nucleic acid sequences of claim 23, wherein the sequence encoding the P2A peptide is set forth in SEQ ID NO:302.

25. The nucleic acid sequences of any one of claims 17-24, wherein the nucleic acid sequences encode the amino acid sequence of SEQ ID NO: 81, 99, 117, 45, 63, 135, 153, 171, 189, 207, 225, 243, 261, 279, 297, or 27.

26. The nucleic acid sequences of any one of claims 20-25, wherein the single expression vector comprises the nucleic acid sequence of SEQ ID NO: 82, 100, 118, 46, 64, 136, 154, 172, 190, 208, 226, 244, 262, 280, 298, or 28.

27. A vector comprising the nucleic acid sequences of any one of claims 17-26.

28. The vector of claim 27, wherein the vector is a viral vector.

29. The vector of claim 28, wherein the viral vector is a lentiviral vector.

30. An engineered cell, comprising the TCR, or antigen-binding fragment thereof, of any one of claims 1-16.

31. An engineered cell, comprising a TCR or antigen-binding fragment thereof, wherein the TCR or antigen-binding fragment thereof is encoded by the nucleic acid sequences of any one of claims 17-26.

32. The engineered cell of claim 30 or 31, wherein the TCR or antigen-binding fragment thereof is heterologous to the cell.

33. The engineered cell of any one of claims 30-32, wherein the engineered cell is derived from a cell line.

34. The engineered cell of any one of claims 30-32, wherein the engineered cell is derived from a primary cell obtained from a subject.

35. The engineered cell of any one of claims 30-34, wherein the engineered cell is a T cell, optionally wherein the T cell is a primary T cell, a natural killer T cell or a cytotoxic T cell.

36. The engineered cell of any one of claims 30-35, wherein expression of one or more endogenous TCR chains of the T cell has been reduced or eliminated, optionally wherein expression of an endogenous TRAC gene and an endogenous TRBC gene of the T cell have been knocked out.

37. The engineered cell of claim 35, wherein the engineered cell has cytotoxic activity against cells expressing CLAVEEVSL (SEQ ID NO:5) or AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule.

38. The engineered cell of claim 36, wherein the MHC molecule is a human leukocyte antigens (HLA)-A molecule.

39. The engineered cell of claim 37, wherein the HLA-A molecule is of serotype HLA-A*02:01, HLA-A*02:06, or HLA-A* 02: 03.

40. The engineered cell of claim 39, wherein the engineered cell has increased cytotoxic activity against cells expressing CLAVEEVSL (SEQ ID NO:5) or AIQDLCLAV (SEQ ID NO: 1) relative to cytotoxic activity against cells expressing AIQDLWQWRKSL (SEQ ID NO:2).

41. A method for producing an engineered cell, comprising introducing the nucleic acid sequences of any one of claims 17-26 or the vector of any one of claims 27-29 into a cell in vitro or ex vivo, optionally wherein the cell is a T cell, optionally wherein the T cell is a primary T cell, a natural killer T cell or a cytotoxic T cell.

42. The method of claim 41, wherein the method further comprises knocking out expression of one or more endogenous TCR genes in the T cell.

43. A composition comprising the TCR or antigen-binding fragment thereof of any one of claims 1-16, the nucleic acid sequences of any one of claims 17-26, or the vector of any one of claims 27-29.

44. A composition comprising the engineered cell of any one of claims 30-40.

45. The composition of claim 43 or 44, further comprising a pharmaceutically acceptable excipient.

46. A method for identifying a T cell receptor (TCR) targeting a NPMlc neoantigen, the method comprising identifying a functional TCR that recognizes the NPMlc neoantigen, among a plurality of functional TCRs, wherein said plurality of functional TCRs are encoded by a plurality of functional TCR-encoding nucleic acid vectors generated by a high-throughput nucleic acid amplification and assembly method using nucleic acid obtained from a single T cell among a plurality of T cells; wherein said plurality of T cells is from donor subject.

47. A method for identifying a T cell receptor (TCR) targeting a NPMlc neoantigen, the method comprising:(i) generating a plurality of functional TCR-encoding nucleic acid vectors by a high-throughput nucleic acid amplification and assembly method using nucleic acids obtained from a single T cell among a plurality of T cells; wherein said T cell is from a donor subject; and(ii) identifying a functional TCR that recognizes the NPMlc neoantigen, among a plurality functional TCRs encoded by the plurality of functional TCR-encoding nucleic acid vectors.

48. The method of claim 46 or 47, wherein the donor subject healthy human donor.

49. The method of any one of claims 46-48, wherein the identified functional TCR recognizes a peptide CLAVEEVSL (SEQ ID NO: 5) or AIQDLCLAV (SEQ ID NO: 1) in the context of an MHC molecule.

50. The method of claim 49, wherein the MHC molecule is a human leukocyte antigens (HLA)-A molecule.

51. The method of claim 50, wherein the HLA-A molecule is of serotype HLA- A*02:01, HLA-A*02:06, or HLA-A* 02: 03.

52. The method of any one of claims 46-51, wherein the T cell from the donor subject is cultured under conditions for cell expansion of the T cell prior to the generating of the plurality of functional TCR-encoding nucleic acid vectors.

53. The method of any one of claims 46-51, wherein the T cell from the donor subject is not cultured under conditions for cell expansion of the T cell prior to the generating of the plurality of functional TCR-encoding nucleic acid vectors.

54. The method of any one of claims 46-53, wherein the high-throughput nucleic acid amplification and assembly method comprises:(1) amplifying a first amplification product and a second amplification product from complementary DNA (cDNA) generated from RNA obtained from the single T cell among the plurality of T cells sorted into each of a plurality of separate locations of a device, wherein: said first amplification product comprises a nucleotide sequence encoding a full- length variable a (Va) region or a full-length variable y (Vy) region of a TCR, and said second amplification product comprises a nucleotide sequence encoding a full-length variable P (VP) region or a full-length variable 5 (V5) region of a TCR; and(2) assembling said first amplification product and said second amplification product from each of said plurality of separate locations into a nucleic acid vector to obtain an assembled nucleic acid vector comprising a nucleotide sequence encoding a functional TCR for each of said plurality of separate locations; and said functional TCR comprises (i) a full-length Va region and a full-length VP region from said single T cell or (ii) a full-length Vy region and a full-length V5 region from said single T cell.

55. An engineered cell, comprising the TCR identified by the method of any one of claims 1-16 or the nucleic acid sequences of any one of claims 17-26.

56. The engineered cell of claim 55, wherein the engineered cell has increased cytotoxic activity against cells expressing CLAVEEVLS (SEQ ID NO:5) or AIQDLCLAV (SEQ ID NO: 1) relative to cytotoxic activity against cells expressing AIQDLWQWRKSL (SEQ ID NO: 2).

57. A composition comprising the engineered cell of claim 55 or 56.

58. The composition of claim 52, further comprising a pharmaceutically acceptable excipient.

59. A method of treatment, the method comprising administering to a subject having a disease or condition, the composition of any one of claims 44, 45, 57, or 58, the engineered T cell of any one of claims 30-40, or an engineered T cell expressing the TCR or antigen-binding fragment thereof of any one of claims 1-16 or the nucleic acid sequences of any one of claims 17-26.

60. A method of treatment, the method comprising administering to a subject having a disease or condition,(a) the TCR or the antigen binding fragment of any one of claims 1-16,(b) a cell expressing the TCR or the antigen binding fragment of any one of claims 1-16,(c) a protein comprising the TCR or the antigen binding fragment of any one of claims 1-16, or(d) a cell expressing the protein comprising the TCR or the antigen binding fragment of any one of claim 1-16,61. The method of claim 59 or 60, wherein the protein comprising the TCR or the antigen binding fragment of any one of claim 1-16 comprises a bispecific T cell engager or a chimeric T cell receptor.

62. The method of any one of claims 59-61, wherein the subject has or has been diagnosed with NPMlc+leukemia, NPMlc+acute myeloid leukemia (AML), or NPMlc+myelodysplastic syndrome (MDS).

63. The method of any one of claims 59-61, wherein the subject has or has been diagnosed a liquid tumor, a hematopoietic tumor, or chronic myeloid leukemia.

64. The method of any one of claims 61-63 wherein the treatment induces or enhances cell death of cells associated with the malignant hematologic disorder, or induces or enhances a graft versus leukemia effect (GVL) in the subject.

65. The method of claim 59 or 90, wherein the disease or condition is a non- malignant disorder or an autoimmune disorder.

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