Treatment and prevention of cancer using HER3 antigen binding molecules

Abstract

The present invention provides the use of an antigen-binding molecule that binds to HER3 for the treatment or prevention of a HER3-related cancer wherein the HER3-related cancer does not comprise a genetic variation that results in an increase in MET expression or an increase in gene product activity.

Description

[0001] This application claims priority to U.S. Provisional Application No. 63 / 457514, filed April 6, 2023, the entire contents and elements of which are incorporated herein by reference for all purposes. Technical Field

[0002] This disclosure relates to the field of molecular biology, and more specifically, to antibody technology and medical treatments and preventative methods. Background Technology

[0003] Many transmembrane protein kinases have been shown to be involved in tumorigenesis (Roskoski Jr 2004). Human epidermal growth factor receptor 3 (HER3), by forming a heterodimer with epidermal growth factor receptor (EGFR) or human epidermal growth factor receptor 2 (HER2), has been identified as a key signaling hub for activating key growth factor signaling pathways such as the MAPK / ERK and PI3K / AKT / mTOR pathways (Gala and Chandarlapaty, Clinical Cancer Research, 2014, Vol. 6: 1410-1416), (Haikala and...). (Clinical Cancer Research, 2021, Vol. 27, No. 13: 3528-3539). HER3 activation has become an important mechanism for tumor progression and acquired resistance to standard therapy in various indications. To date, HER3-targeting approaches have not shown the expected clinical efficacy. Insufficient inhibition of HER3-mediated signaling is one possible explanation.

[0004] WO2023 / 017151A1 discloses methods for treating and preventing cancers characterized by the presence or absence of mutations in genes encoding HER3-mediated signaling factors. Summary of the Invention

[0005] In a first aspect, this disclosure provides an antigen-binding molecule that binds to HER3 for use in treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers do not contain genetic variations that lead to increased MET expression or enhanced activity of gene products.

[0006] This disclosure also provides the use of antigen-binding molecules that bind to HER3 for the preparation of medicaments for the treatment or prevention of HER3-related cancers in subjects, wherein the HER3-related cancers do not contain genetic variations that lead to increased MET expression or enhanced activity of gene products.

[0007] This disclosure also provides a method for treating or preventing HER3-related cancers in a subject, comprising administering to the subject a therapeutically or preventively effective amount of an antigen-binding molecule that binds to HER3, wherein the HER3-related cancers do not contain genetic variations that lead to increased MET expression or enhanced activity of gene products.

[0008] According to various aspects of this disclosure, in some embodiments, the HER3-related cancers: (i) do not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity; or (ii) do not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity (e.g., do not contain activating mutations of PIK3CA); or (iii) do not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity; or (iv) do not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity.

[0009] In some embodiments, the HER3-related cancers: (i) do not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity; and (ii) do not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity (e.g., do not contain activating mutations of PIK3CA); and (iii) do not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity; and (iv) do not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity.

[0010] In some implementations, the HER3-related cancers: (i) do not contain MET gene amplification; and (ii) do not contain KRAS activating mutations; and (iii) do not contain PIK3CA activating mutations; and (iv) do not contain BRAF activating mutations; and (v) do not contain PTEN gene deletions.

[0011] In some embodiments, the HER3-related cancers: (i) lack MET gene amplification; and (ii) are homozygous for the KRAS wild-type allele; and (iii) are homozygous for the PIK3CA wild-type allele; and (iv) are homozygous for the BRAF wild-type allele; and (v) are homozygous for the PTEN wild-type allele.

[0012] In some embodiments, the HER3-related cancer comprises amplification of one or more genes located on the long arm of chromosome 3. In some embodiments, the HER3-related cancer comprises amplification of one or more genes located on chromosome 3 3q26-3q28. In some embodiments, the HER3-related cancer comprises amplification of one or more genes selected from the following: TP63, SOX2, and PIK3CA.

[0013] In some embodiments, the HER3-related cancer comprises amplification of one or more genes located on the short arm of chromosome 7. In some embodiments, the HER3-related cancer comprises amplification of one or more genes located on chromosome 7p11. In some embodiments, the HER3-related cancer comprises EGFR gene amplification.

[0014] In some embodiments, the HER3-related cancer comprises the deletion of one or more genes located on the short arm of chromosome 3. In some embodiments, the HER3-related cancer comprises the deletion of one or more genes located on chromosome 3p21. In some embodiments, the HER3-related cancer comprises the deletion of the TUSC2 gene.

[0015] In some implementations, the HER3-related cancers include genetic variations that lead to increased expression of the HER3 ligand.

[0016] In some implementations, the HER3-related cancers include NRG gene fusions, NRG1 gene fusions, or NRG2 gene fusions.

[0017] In some embodiments, the HER3-related cancer comprises an NRG gene fusion selected from the following: CLU-NRG1, CD74-NRG1, DOC4-NRG1, SLC3A2-NRG1, RBPMS-NRG1, WRN-NRG1, SDC4-NRG1, RAB2IL1-NRG1, VAMP2-NRG1, KIF13B-NRG1, THAP7-NRG1, SMAD4-NRG1, MDK-NRG1, TNC-NRG1, DIP2B-NRG1, MR PL13-NRG1, PARP8-NRG1, ROCK1-NRG1, DPYSL2-NRG1, ATP1B1-NRG1, CDH6-NRG1, APP-NRG1, AKAP13-NRG1, THBS1-NRG1 , FOXA1-NRG1, PDE7A-NRG1, RAB3IL1-NRG1, CDK1-NRG1, BMPRIB-NRG1, TNFRSF10B-NRG1, MCPH1-NRG1 and SLC12A2-NRG2.

[0018] This disclosure also provides a HER3-binding antigen-binding molecule for use in a method of treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers comprise amplifications of one or more genes located on the long arm of chromosome 3. This disclosure also provides the use of the HER3-binding antigen-binding molecule for preparing a medicament for treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers comprise amplifications of one or more genes located on the long arm of chromosome 3. This disclosure also provides a method of treating or preventing HER3-related cancers in a subject, comprising administering to the subject a therapeutically or preventively effective amount of the HER3-binding antigen-binding molecule, wherein the HER3-related cancers comprise amplifications of one or more genes located on the long arm of chromosome 3. In some embodiments, the one or more genes located on the long arm of chromosome 3 are located within chromosome 3q26-3q28. In some embodiments, the one or more genes are selected from TP63, SOX2, and PIK3CA.

[0019] This disclosure also provides a HER3-binding antigen-binding molecule for use in a method of treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers comprise amplifications of one or more genes located on the short arm of chromosome 7. This disclosure also provides the use of the HER3-binding antigen-binding molecule for preparing a medicament for treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers comprise amplifications of one or more genes located on the short arm of chromosome 7. This disclosure also provides a method of treating or preventing HER3-related cancers in a subject, comprising administering to the subject a therapeutically or preventively effective amount of the HER3-binding antigen-binding molecule, wherein the HER3-related cancers comprise amplifications of one or more genes located on the short arm of chromosome 7. In some embodiments, the one or more genes located on the short arm of chromosome 7 are located within chromosome 7p11. In some embodiments, the gene is EGFR.

[0020] This disclosure also provides a HER3-binding antigen-binding molecule for use in a method of treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers comprise the deletion of one or more genes located on the short arm of chromosome 3. This disclosure also provides the use of the HER3-binding antigen-binding molecule for preparing a medicament for treating or preventing HER3-related cancers in a subject, wherein the HER3-related cancers comprise the deletion of one or more genes located on the short arm of chromosome 3. This disclosure also provides a method of treating or preventing HER3-related cancers in a subject, comprising administering to the subject a therapeutically or preventively effective amount of the HER3-binding antigen-binding molecule, wherein the HER3-related cancers comprise the deletion of one or more genes located on the short arm of chromosome 3. In some embodiments, the one or more genes located on the short arm of chromosome 3 are located within chromosome 3p21. In some embodiments, the gene is TUSC2.

[0021] According to various aspects of this disclosure, in some embodiments, the HER3-related cancer comprises an amplification of one or more genes selected from the following: TP63, SOX2, and PIK3CA.

[0022] In some implementations, the HER3-related cancer includes TP63 gene amplification, SOX2 gene amplification, and PIK3CA gene amplification.

[0023] This disclosure also provides an antigen-binding molecule that binds to HER3 for use in treating or preventing HER3-related cancers in subjects, wherein the HER3-related cancers comprise genetic variations that lead to increased MET expression or enhanced activity of gene products, and the method further includes administering an antagonist of HER3-mediated signaling.

[0024] This disclosure also provides the use of antigen-binding molecules that bind to HER3 for the preparation of medicaments for the treatment or prevention of HER3-related cancers in subjects, wherein the HER3-related cancers comprise genetic variations that lead to increased MET expression or enhanced activity of gene products, and the treatment methods further include administration of an antagonist of HER3-mediated signaling.

[0025] This disclosure also provides a method for treating or preventing HER3-related cancers in a subject, comprising administering to the subject a therapeutically or preventively effective amount of an antigen-binding molecule that binds to HER3, wherein the HER3-related cancers contain genetic variations that result in increased MET expression or enhanced activity of gene products, and the method further comprises administering an antagonist of HER3-mediated signaling.

[0026] According to various aspects of this disclosure, the HER3-related cancers: (i) contain genetic variants that lead to increased KRAS expression or enhanced gene product activity; or (ii) contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity; or (iii) contain genetic variants that lead to increased BRAF expression or enhanced gene product activity; or (iv) contain genetic variants that lead to decreased PTEN expression or reduced gene product activity.

[0027] In some embodiments, the HER3-related cancers include: (i) genetic variants that lead to increased KRAS expression or enhanced gene product activity; (ii) genetic variants that lead to increased PIK3CA expression or enhanced gene product activity; (iii) genetic variants that lead to increased BRAF expression or enhanced gene product activity; and (iv) genetic variants that lead to decreased PTEN expression or reduced gene product activity.

[0028] This disclosure also provides a method for screening subjects suitable for antigen-binding molecular therapy that binds to HER3, comprising:

[0029] (a) Analyze the subject's cancer to determine whether the cancer contains genetic variations that lead to increased MET expression or enhanced gene product activity; and

[0030] (b) If step (a) determines that the subject’s cancer does not contain such genetic variants, then the subject is selected to receive antigen-binding molecule therapy that binds to HER3.

[0031] In some implementations, the method includes:

[0032] (a)

[0033] (i) Analyze the subject's cancer to determine whether the cancer contains genetic variations that lead to increased KRAS expression or enhanced gene product activity; or

[0034] (ii) Analyze the subject's cancer to determine whether the cancer contains a PIK3CA activating mutation; or

[0035] (iii) Analyze the subject's cancer to determine whether the cancer contains genetic variations that lead to increased BRAF expression or enhanced gene product activity; or

[0036] (iv) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to reduced PTEN expression.

[0037] Or genetic variations that reduce the activity of gene products; and

[0038] (b) If step (a) determines that the subject’s cancer does not contain such genetic variants / mutations, then the subject is selected to receive antigen-binding molecule therapy that binds to HER3.

[0039] In some implementations, the method includes:

[0040] (a)

[0041] (i) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to increased KRAS expression or

[0042] Genetic variations that enhance the activity of gene products; and

[0043] (ii) Analyze the subject's cancer to determine whether the cancer contains a PIK3CA activating mutation; and

[0044] (iii) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to increased BRAF expression.

[0045] Or genetic variations that enhance the activity of gene products; and

[0046] (iv) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to reduced PTEN expression.

[0047] Or genetic variations that reduce the activity of gene products; and

[0048] (b) If step (a) determines that the subject’s cancer does not contain such genetic variants / mutations, then the subject is selected to receive antigen-binding molecule therapy that binds to HER3.

[0049] In some implementations, the method further includes:

[0050] (c) Administer the HER3-binding antigen-binding molecule to the selected treatment subject from step (b).

[0051] This disclosure also provides a method for screening subjects suitable for (i) HER3-mediated signaling antagonists and (ii) HER3-binding antigen-binding molecules, comprising:

[0052] (a) Analyze the subject's cancer to determine whether the cancer contains genetic variations that lead to increased MET expression or enhanced gene product activity; and

[0053] (b) If step (a) determines that the subject’s cancer contains such a genetic variant, then the subject is selected to receive (i) a HER3-mediated signaling antagonist and (ii) a HER3-binding antigen-binding molecule.

[0054] In some implementations, the method includes:

[0055] (a)

[0056] (i) Analyze the subject's cancer to determine whether the cancer contains genetic variations that lead to increased KRAS expression or enhanced gene product activity; or

[0057] (ii) Analyze the subject's cancer to determine whether the cancer contains a PIK3CA activating mutation; or

[0058] (iii) Analyze the subject's cancer to determine whether the cancer contains genetic variations that lead to increased BRAF expression or enhanced gene product activity; or

[0059] (iv) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to reduced PTEN expression.

[0060] Or genetic variations that reduce the activity of gene products; and

[0061] (b) If step (a) determines that the subject’s cancer contains such genetic variants / mutations, then the subject is selected to receive (i) a HER3-mediated signaling antagonist and (ii) a HER3-binding antigen-binding molecule.

[0062] In some implementations, the method includes:

[0063] (a)

[0064] (i) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to increased KRAS expression or

[0065] Genetic variations that enhance the activity of gene products; and

[0066] (ii) Analyze the subject's cancer to determine whether the cancer contains a PIK3CA activating mutation; and

[0067] (iii) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to increased BRAF expression.

[0068] Or genetic variations that enhance the activity of gene products; and

[0069] (iv) Analyze the subjects' cancers to determine whether the cancers contain substances that lead to reduced PTEN expression.

[0070] Or genetic variations that reduce the activity of gene products; and

[0071] (b) If step (a) determines that the subject’s cancer contains such genetic variants / mutations, then the subject is selected to receive (i) a HER3-mediated signaling antagonist and (ii) a HER3-binding antigen-binding molecule.

[0072] In some implementations, the method further includes:

[0073] (c) Administer (i) a HER3-mediated signal transduction antagonist and (ii) an antigen-binding molecule that binds to HER3 to the selected subjects in step (b).

[0074] According to various aspects of this disclosure, in some embodiments, the HER3-related cancers are selected from: solid tumors, breast cancer, breast carcinoma, ductal carcinoma, gastric cancer, gastric carcinoma, gastric adenocarcinoma, colorectal cancer, colorectal carcinoma, colorectal adenocarcinoma, head and neck cancer, head and neck squamous cell carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, ovarian cancer, ovarian carcinoma, ovarian serous adenocarcinoma, renal cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, renal papillary cell carcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic duct adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, esophageal cancer, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, bile duct cancer, uterine cancer, endometrial cancer of the uterine corpus, thyroid cancer. (carcinoma), pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma, and thymoma.

[0075] In some embodiments, the antigen-binding molecule that binds to HER3 is selected from: 10D1F, seribantumab, elgemtumab, patritumab, GSK2849330, lumretuzumab, CDX-3379, AV-203, barecetamab, TK-A3, TK-A4, MP-EV20, 1A5-3D4, 9F7-F11, 16D3-C1, NG33, A5, F4, huHER3-8, REGN1400, and zenocutuzumab.

[0076] In some embodiments, the HER3-binding antigen-binding molecule comprises:

[0077] (i) Heavy chain variable (VH) regions containing the following CDRs:

[0078] HC-CDR1 has the amino acid sequence of SEQ ID NO:40;

[0079] HC-CDR2 has the amino acid sequence of SEQ ID NO:43;

[0080] HC-CDR3, having the amino acid sequence of SEQ ID NO:48; and

[0081] (ii) Light chain variable (LH) regions containing the following CDRs:

[0082] LC-CDR1 has the amino acid sequence of SEQ ID NO:66;

[0083] LC-CDR2 has the amino acid sequence of SEQ ID NO:69;

[0084] LC-CDR3 has the amino acid sequence of SEQ ID NO:74.

[0085] In some embodiments, the HER3-binding antigen-binding molecule comprises:

[0086] (i) VH region containing the following CDRs:

[0087] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0088] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0089] HC-CDR3, having the amino acid sequence of SEQ ID NO:45; and

[0090] (ii) VL regions containing the following CDRs:

[0091] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0092] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0093] LC-CDR3 has the amino acid sequence of SEQ ID NO:70.

[0094] In some embodiments, the antigen-binding molecule comprises:

[0095] The VH region contains an amino acid sequence that has at least 70% sequence identity with the amino acid sequence of SEQ ID NO:33; and

[0096] The VL region contains an amino acid sequence that has at least 70% sequence identity with the amino acid sequence of SEQ ID NO:58.

[0097] In some embodiments, the antigen-binding molecule comprises:

[0098] A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:75; and

[0099] A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:76.

[0100] According to various aspects of this disclosure, in some embodiments, the method of treating or preventing HER3-related cancers further includes administering an EGFR antagonist to the subject.

[0101] In some embodiments, the EGFR antagonist is an antigen-binding molecule that binds to EGFR.

[0102] In some embodiments, the EGFR-binding antigen-binding molecule comprises:

[0103] (i) VH region containing the following CDRs:

[0104] HC-CDR1 has the amino acid sequence of SEQ ID NO:78;

[0105] HC-CDR2 has the amino acid sequence of SEQ ID NO:79;

[0106] HC-CDR3, having the amino acid sequence of SEQ ID NO:80; and

[0107] (ii) VL regions containing the following CDRs:

[0108] LC-CDR1 has the amino acid sequence of SEQ ID NO:82;

[0109] LC-CDR2 has the amino acid sequence of SEQ ID NO:83;

[0110] LC-CDR3 has the amino acid sequence of SEQ ID NO:84.

[0111] In some embodiments, the antigen-binding molecule that binds to EGFR includes:

[0112] The VH region contains an amino acid sequence that has at least 70% sequence identity with the amino acid sequence of SEQ ID NO:77; and

[0113] The VL region contains an amino acid sequence that has at least 70% sequence identity with the amino acid sequence of SEQ ID NO:81.

[0114] In some embodiments, the antigen-binding molecule that binds to EGFR includes:

[0115] A polypeptide comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:85; and

[0116] A polypeptide comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:86. Detailed Implementation

[0117] This disclosure is partly based on the inventors' discovery that cancers containing genetic variants that lead to increased MET expression (especially MET amplification) may respond poorly to anti-HER3 antibody therapy. Cancers lacking such genetic variants are identified as cancers sensitive to anti-HER3 antibody therapy. The inventors confirmed that, in addition to assessing PTEN, KRAS, PIK3CA, and BRAF mutation status as described in WO2023 / 017151A1, further detection of MET amplification status can predict a positive response to anti-HER3 antibody therapy with higher sensitivity (see Example 4).

[0118] This disclosure is also based on the inventors' discovery that cancers containing genetic variations (particularly amplification of such genes) that lead to increased expression of genes located on chromosome 3 3q26-3q28 may respond better to anti-HER3 antibody therapy. Cancers with such genetic variations are identified as cancers sensitive to anti-HER3 antibody therapy.

[0119] HER3 and HER3-mediated signal transduction

[0120] HER3 (also known as ERBB3, LCCS2, MDA-BF-1) is a protein identified by UniProt P21860.

[0121] The structure and function of HER3 are described in Cho and Leahy, Science, 2002, Vol. 297, No. 5585, pp. 1330-1333; Singer et al., Journal of Biological Chemistry, 2001, Vol. 276, pp. 44266-44274; Roskoski et al., Pharmacol. Res., 2014, Vol. 79, pp. 34-74; Bazley and Gullick, Endocrine-Related Cancer, 2005, pp. 17-27; and Mujoo et al., Oncotarget, 2014, Vol. 5, No. 21, pp. 10222-10236. All of these references are incorporated into this article by way of citation. HER3 is a single-transmembrane ErbB receptor tyrosine kinase with an N-terminal extracellular region (SEQ ID NO:9) containing two leucine-rich subdomains (domains I and III, SEQ ID NO:15 and 17, respectively) and two cysteine-rich subdomains (domains II and IV, SEQ ID NO:16 and 18, respectively). Domain II contains a β-hairpin dimer loop (SEQ ID NO:19), which participates in intermolecular interactions with other HER receptor molecules. The extracellular region is connected to the cytoplasmic region (SEQ ID NO:11) via a transmembrane region (SEQ ID NO:10). The cytoplasmic region contains a juxtamembrane segment (SEQ ID NO:12), a protein kinase domain (SEQ ID NO:13), and a C-terminal segment (SEQ ID NO:14).

[0122] In this specification, "HER3" refers to HER3 from any species, including HER3 isotypes, fragments, variants (including mutants) or homologs from any species.

[0123] As used herein, a “fragment,” “variant,” or “homolog” of a protein may optionally be defined as a molecule having at least 60%, preferably 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity with a reference protein (such as a reference isotype). In some embodiments, fragments, variants, isotypes, and homologs of a reference protein may be characterized by their ability to perform the functions performed by the reference protein.

[0124] A “fragment” typically refers to a portion of a reference protein. A “variant” typically refers to a protein whose amino acid sequence contains one or more amino acid substitutions, insertions, deletions, or other modifications relative to the reference protein, but still maintains a degree of sequence identity (e.g., at least 60%) comparable to the reference protein's amino acid sequence. A “homotype” typically refers to a variant of a reference protein expressed by a species of the same species as the reference protein (e.g., human HER3 isotypes 1 through 5, which are isotypes of each other). A “homolog” typically refers to a variant of a reference protein expressed by a species of a different species than the reference protein. For example, human HER3 isotype 1 (P21860-1, v1; SEQ ID NO: 1) and rhesus monkey HER3 (UniProt: F7HEH3-1, v2; SEQ ID NO: 20) are homologs. Homologs include orthologs.

[0125] The “fragment” of the reference protein may have any length (in terms of amino acid number), but may optionally be at least 20% of the length of the reference protein (i.e., the protein from which the fragment is derived), and the maximum length may be one of 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the length of the reference protein.

[0126] The minimum length of the HER3 fragment can be any of 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, or 1200 amino acids, and the maximum length can be any of 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1200, or 1300 amino acids.

[0127] In some embodiments, the HER3 is a mammalian HER3 (such as primate HER3 (rhesus monkey, cynomolgus monkey, non-human primate or human) and / or rodent HER3 (such as rat or mouse). Isotypes, fragments, variants, or homologs of HER3 are optionally defined as having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with an immature or mature HER3 isotype of a particular species (such as human).

[0128] The isotype, fragment, variant, or homolog may optionally be a functional isotype, fragment, variant, or homolog, such as having the functional properties / activities of a reference HER3 (e.g., human HER3 subtype 1), which can be analyzed by appropriate detection methods. For example, an isotype, fragment, variant, or homolog of HER3 may exhibit associations with one or more of the following: HER2, NRG1 (types I, II, III, IV, V, or VI), or NRG2 (α or β type).

[0129] In some embodiments, HER3 comprises or consists of an amino acid sequence that is identical to any one of SEQ ID NO:1 to 8 having an amino acid sequence identity of at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0130] In some embodiments, the HER3 fragment comprises or consists of an amino acid sequence that is identical to any one of SEQ ID NO: 9 to 19, for example, one of 9, 16 or 19, having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% of the amino acid sequence.

[0131] HER3 signaling involves receptor heterodimerization (with other ErBB receptors, such as HER2 and EGFR) and subsequent autophosphorylation via a protein kinase domain of tyrosine residues in the cytoplasm. HER3 lacks kinase activity and cannot form stable homodimers. Therefore, HER3 must initiate signal transduction by binding to a kinase-active heterodimer chaperone (such as EGFR or HER2) (Berger MB et al., FEBS Lett, 2004, Vol. 569, pp. 332-6; Kim HH et al., Biochem J, 1998, Vol. 334, pp. 189-95).

[0132] Multimerization (e.g., dimerization) of HER receptor family members is essential for activating cell growth signaling pathways. HER3 can dimerize with other HER family members in both ligand-dependent and ligand-independent ways. The HER3 extracellular domain (ECD) exists in a reversible equilibrium between a closed, inactive conformation and an open, active conformation, in which the dimerizing arms within domain II are exposed to allow dimerization along the domain II dimerization interface, particularly through the cysteine-rich CR1 region (Carraway KL et al., *Nature*, 1997, Vol. 387, pp. 512-6; Riese DJ et al., *Molecular and Cell Biology*, 1995, Vol. 15, pp. 5770-6; Harari D et al., *Oncogene*, 1999, Vol. 18, pp. 2681-9; Zhang D et al., *Proceedings of the National Academy of Sciences of the United States of America*, 1997, Vol. 94, pp. 9562-7; Meyer et al., *Nature*, 1995, Vol. 378, pp. 386-90; Jura N et al., *Proceedings of the National Academy of Sciences of the United States of America*). (USA) 2009, Vol. 106, pp. 21608-13; Fornaro L et al., *Nature Reviews Gastroenterology & Hepatology*, Vol. 8, 2011, pp. 369-83; Mota et al., *Oncotarget*, Vol. 5, 2015, pp. 89284-306). HER3 is "activated" when the equilibrium tilts towards the open conformation, increasing the probability of forming an active heterodimer. The traditional activation model is ligand-dependent, meaning that the equilibrium shifts when HER3 in the open conformation is stabilized by binding to its ligands, such as neuroregulatory proteins (NGRs), namely NRG1 (also known as regulatory protein / HRG) or NRG2. Furthermore, when any dimerizing chaperone reaches a sufficient concentration, it will drive the equilibrium towards the open conformation because it binds to and temporarily stabilizes HER3 in the open conformation. It is referred to as ligand-independent activation (Jura N et al., Proc Natl Acad Sci USA, 2009, Vol. 106, pp. 21608-13; Fornaro L et al., Nat Rev Gastroenterol Hepatol, 2011, Vol. 8, pp. 369-83; Mota et al., Oncotarget, 2015, Vol. 5, pp. 89284-306).

[0133] In this article, "HER3-mediated signaling" refers to signal transduction mediated by HER3 and / or HER3-containing multimeric ErBB family member receptor complexes. "Signal transduction" refers to signal transduction and other cellular processes that regulate cellular activity. HER3-mediated signaling can be mediated by HER3-containing receptor complexes, such as heteropolymeric complexes containing HER3 and other HER receptors (e.g., HER2, EGFR). HER3-mediated signaling can be ligand-dependent, such as triggered by NGR (NRG1 or NRG2) binding, or ligand-independent.

[0134] HER3-mediated signaling is transmitted intracellularly through the MAPK / ERK and PI3K / AKT / mTOR pathways to promote cell survival and proliferation. HER3-mediated signaling is described in Gala and Chandarlapaty, *Clinical Cancer Research*, 2014, Vol. 20, No. 6, pp. 1410-1416; Mishra et al., *Oncol Rev.*, 2018, Vol. 12, No. 1, p. 355; Baselga et al., *Nature Rev. Cancer*, 2009, Vol. 9, pp. 463-475; Yarden et al., *Nature Rev. Molecular Cell Biology*, 2001, Vol. 2, pp. 350-373; Mota et al., *Oncotarget*, 2015, Vol. 5, pp. 89284-89306; and Haikala and... These references are all incorporated into this article by way of citation from Clin. Cancer Res., 2021, Vol. 27, pp. 3528-3539.

[0135] In receptor complexes containing HER3, phosphorylated tyrosine residues in the protein kinase domain recruit the aptamer / effect protein GRB2 through interaction with its SH2 domain. Upon ligand stimulation, the activated receptor (EGFR / HER2) undergoes autophosphorylation, providing phosphotyrosine residues for GRB2 recruitment. GRB2 binds to the guanine nucleotide exchange factor SOS via its SH3 domain. Activated SOS in the GRB2-SOS complex promotes GDP removal, thereby activating Ras family GTPases, such as H-Ras, N-Ras, and K-Ras. Activated Ras GTPases then activate RAF kinases, such as A-Raf, B-Raf, and C-Raf. RAF kinases then phosphorylate and activate MEK1 and MEK2, which in turn rephosphorylate and activate MAPK (also known as ERK). Activated MAPK can directly regulate the activity of transcription factors such as c-Myc. Activated MAPK can also upregulate mRNA translation into protein via RSK phosphorylation, subsequently phosphorylating and activating the 40S ribosomal protein S6. Activated MAPK can also phosphorylate and activate MNK, which in turn phosphorylates and activates the transcription factor CREB.

[0136] Phosphorylated tyrosine residues in the HER3 protein kinase domain can also recruit the p85 subunit of PI3K through its SH2 domain. Binding to p85 induces allosteric activation of the p110α subunit of the PI3K lipid kinase. Activated PI3K leads to the conversion of PIP2 to PIP3, which recruits AKT and is phosphorylated and activated by mTORC2 and PDK1. Phosphorylated AKT possesses various activities, including activation of CREB and mTOR. PTEN antagonizes the PI3K / AKT / mTOR pathway signaling by dephosphorylating PIP3 to PIP2, and PP2A inhibits the PI3K / AKT / mTOR pathway by dephosphorylating AKT.

[0137] Oncogenic Src homology region 2 protein tyrosine phosphatase 2 (SHP2) can promote tumor progression and serves as a key hub connecting multiple oncogenic signaling pathways, such as PI3K / Akt and Ras / Raf / MAPK (Dong et al., Frontiers in Cell and Developmental Biology, March 11, 2021). GAB2 binds to GRB2 and phosphorylates multiple tyrosine residues, thereby enabling it to bind to the SH2 domains of SHP2 and p85 (Adams et al., Molecular Cancer Research, October 2012, Vol. 10, No. 10, pp. 1265-70; Liu et al., Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, pp. 984-989, 2016). These interactions induce conformational changes that alleviate the self-inhibition of the SHP2 catalytic site (Neel et al., Trends in Biochemistry, June 2003, Vol. 28, No. 6, pp. 284-93) and the inhibition of the p110 catalytic subunit in PI3K by p85 (Cuevas et al., Journal of Biochemistry, July 20, 2001, Vol. 276, No. 29, pp. 274-55-6). Studies have shown that SHP2 can activate RAS by directly dephosphorylating RAS (Bunda et al., Nature Communications, November 30, 2015, Vol. 6, No. 8859), inhibiting RASGAP (RAS GTPase activator protein) (Neel et al., Trends in Biochemistry, June 2003, Vol. 28, No. 6, pp. 284-93), and inhibiting SPRY (Hanafusa et al., Journal of Biochemistry, May 28, 2004, Vol. 279, No. 22, pp. 22992-5). SHP2 overexpression has been shown to enhance tumor invasion by activating the PI3K / Akt axis (Hu et al., Onco Targets Ther, 2017, Vol. 10, pp. 3881-3891), while SHP2 knockdown inhibits cell migration, and the tumor-promoting effect of SHP2 is partially related to the Akt signaling pathway (Cao et al., Pathol. Res. Pract., 2019, Vol. 215, pp. 152621).

[0138] STAT3 and STAT5 proteins are transcription factors that enhance the expression of p85α, p110α, and AKT1, thereby enhancing signal transduction through the PI3K / AKT signaling cascade (Radler et al., *Molecular Cell Endocrinology*, August 15, 2017, Vol. 451, pp. 31-39). HER3-mediated STAT3 activation leads to upregulation of SOX2 expression, promoting survival. Upon activation by JAK2, phosphorylated STAT5 binds to the SH2 domain of the PI3K p85α regulatory subunit in a PRL-dependent manner, suggesting that STAT5 may directly participate in the signal transduction of the PI3K complex. Another phosphorylated EGFR kinase is the cytokine-regulated tyrosine kinase Jak2, therefore the MAPK pathway can still be activated even in EGFR kinase-deficient mutants (Mishra et al., *Oncol Rev.*, 2018, Vol. 12, No. 1, p. 355; Baselga et al., *Nature Reviews Cancer*, 2009, Vol. 9, pp. 463-75). Through comprehensive observation of genetic models with overexpression or lack of STAT5 and AKT activity, or genetic models expressing PTEN mutants, it has been confirmed that the functions of STAT5 as a survival factor in normal breast development and as an oncogene in breast cancer development are mediated through the PI3K / AKT pathway (Radler et al., *Molecular Cell Endocrinol.*, August 15, 2017, Vol. 451, pp. 31-39).

[0139] Studies have shown that the proto-oncogene MET can phosphorylate and activate HER3. (See Frazier et al., *Oncogene*, 2019, Vol. 38, No. 11, pp. 1936-1950). The study indicates that increased MET expression enhances HER3 phosphorylation. HER3 preferentially interacts with MET during maturation in the secretory pathway, leading to the accumulation of phosphorylated HER3 in the Golgi apparatus.

[0140] In certain cancers, the PI3K / AKT signaling pathway is frequently altered due to gene amplification and / or mutation. The TP63 and SOX2 cell line genes are also present in the frequently amplified 3q26 to 3q28 chromosomal region (the region containing the PIK3CA gene). TP63, a member of the TP53 gene family, is expressed in the basal cell compartments of the skin, esophagus, lung airways, and larynx during development and homeostasis. TP63 has been used as a diagnostic marker for the differential diagnosis of lung squamous cell carcinoma from lung adenocarcinoma, and esophageal squamous cell carcinoma from esophageal adenocarcinoma. Preclinical data show that TP63 regulates NRG1 expression in SCC (see Hedge et al., eLife, 2019, Vol. 8, e46551), indicating that the HER3 signaling pathway is active in TP63-amplified squamous cell carcinoma.

[0141] Genetic variation, alleles and genotype

[0142] Various aspects and embodiments of this disclosure relate to therapeutic and preventive interventions for treating / preventing cancers characterized by the absence or presence of genetic variations in gene-encoding factors associated with HER3-mediated signaling.

[0143] In particular, this disclosure relates to cancers characterized by: the absence or presence of genetic variations that result in increased or decreased expression levels of genes and / or proteins encoding gene factors associated with HER3-mediated signaling, or genetic variations that result in increased or decreased activity of gene products of gene editing factors associated with HER3-mediated signaling.

[0144] The genetic variations presupposed in this disclosure include mutations, gene amplifications, and gene deletions.

[0145] As used herein, a “mutation” refers to a difference relative to a reference nucleotide sequence (e.g., the most common nucleotide sequence of a given gene, which may be referred to as the “wild-type” nucleotide sequence). In some embodiments, a mutation may be or include one or more of the following relative to a reference sequence (e.g., a wild-type sequence): nucleotide polymorphisms (e.g., single nucleotide polymorphisms (SNPs) or multiple nucleotide polymorphisms (MNPs)), insertions, deletions, frameshift mutations, missense mutations, or translocations.

[0146] Nucleotide polymorphisms can include replacing a nucleotide in a reference sequence with another different nucleotide (such as an SNP), or can include replacing two or more nucleotides in a reference sequence with different nucleotides (such as in the case of an MNP). An insertion is the introduction of one or more nucleotides into the nucleotide sequence of a reference sequence. A deletion is the removal / removal of one or more nucleotides from the nucleotide sequence of a reference sequence. A frameshift mutation is an insertion or deletion mutation that alters the reading frame of the RNA encoded by the mutated nucleotide sequence. Such frameshift mutations include insertions / deletions of nucleotides not divisible by 3. A missense mutation is a substitution (or insertion or deletion) that does not change the amino acid sequence of the peptide / polypeptide translated from the RNA encoded by the mutated nucleotide sequence. This mutation can result in the substitution of one or more nucleotides with different nucleotides, where the codon containing the non-substituted nucleotide and the codon containing the substituted nucleotide encode the same amino acid (as a result of genetic code degeneracy). Other such missense mutations can exist in non-protein-coding regions of a nucleotide sequence, such as introns. A translocation is the large-scale substitution of a nucleotide or nucleotide sequence in a reference sequence with different nucleotide / nucleotide sequences. Translocation is the result of recombination between nucleotide sequences, such as occurring in different chromosomes.

[0147] As used herein, an "activating mutation" refers to a known or predicted mutation that results in an increase in the expression level of an associated gene (gene or protein), and / or a known or predicted mutation that results in enhanced activity of a gene product. Activating mutations can produce one or more of the following in cells comprising one or more alleles of a gene containing the activating mutation: increased gene transcription; increased levels of gene-encoded RNA; decreased degradation of gene-encoded RNA; increased levels of gene product; increased levels of gene-encoded peptides / peptides; increased levels of normal splicing of pre-mRNA encoded by the gene; increased translation of gene-encoded peptide / peptide mRNA; increased levels of normal post-translational processing of gene-encoded peptides / peptides; increased levels of normal transport of gene-encoded peptides / peptides; decreased degradation of gene-encoded peptides / peptides; increased levels of one or more functional properties exhibited by the gene product; increased levels of one or more functional properties exhibited by gene-encoded peptides / peptides; new functional properties exhibited by the gene product; and / or new functional properties exhibited by gene-encoded peptides / peptides. In some embodiments, the activating mutation as described in this disclosure is not or does not include an amplification of the associated gene.

[0148] Conversely, an "inactivating mutation" refers to a mutation that results in a reduced expression level of a given gene, and / or a reduced activity of a gene product. Inactivating mutations can produce one or more of the following in cells comprising one or more alleles of a gene containing the inactivating mutation: reduced gene transcription; reduced levels of gene-encoded RNA; increased degradation of gene-encoded RNA; reduced levels of gene product; reduced levels of gene-encoded peptides / polypeptides; reduced levels of normal splicing of pre-mRNA encoded by the gene; reduced translation of mRNA encoding peptides / polypeptides; reduced levels of normal post-translational processing of gene-encoded peptides / polypeptides; reduced levels of normal transport of gene-encoded peptides / polypeptides; increased degradation of gene-encoded peptides / polypeptides; decreased levels of one or more functional properties exhibited by the gene product; and / or decreased levels of one or more functional properties exhibited by gene-encoded peptides / polypeptides. In some embodiments, the inactivating mutation described herein is not or does not include the deletion of the relevant gene.

[0149] As used herein, "amplification" refers to an increase in the copy number of a given gene or a fragment thereof. Therefore, "amplification" of a given reference gene means the presence of one or more additional copies of the given gene or an additional copy of the given gene fragment relative to the copy number of the gene / fragment present in a healthy (e.g., non-cancerous) diploid cell. Thus, a cell containing an amplification of a given reference gene includes one or more additional copies of the given gene or an additional copy of the given gene fragment relative to the copy number of the gene / fragment present in a healthy (e.g., non-cancerous) diploid cell. In some embodiments, a cell containing an amplification of a given reference gene / fragment includes two or more copies of the relevant gene / fragment. In some embodiments, the cell includes 3, 4, 5, 6, 7, 8, 9, 10 or more copies of the relevant gene / fragment.

[0150] The amplifications described in this disclosure include “gain,” which is sometimes used in the art to describe a relatively small increase in the copy number of a given gene or a fragment thereof (e.g., 3 or 4 copies of the relevant gene / fragment).

[0151] As used herein, "gene deletion" refers to a reduction in the copy number of a given gene or a fragment thereof. Therefore, "deletion" of a given reference gene means a reduction in the copy number of the given gene or a reduction in the copy number of a given gene fragment relative to the copy number of the gene / fragment present in a healthy (e.g., non-cancerous) diploid cell. Thus, a cell containing a deletion of a given reference gene has a reduced copy number of the given gene or a reduced copy number of the given gene fragment relative to the copy number of the gene / fragment present in a healthy (e.g., non-cancerous) diploid cell. In some embodiments, a cell containing a deletion of a given reference gene / fragment contains fewer than two copies of the relevant gene / fragment. In some embodiments, the cell contains 0 or 1 copies of the relevant gene / fragment.

[0152] Gene deletions as described in this disclosure include "shallow" and "deep" deletions. A shallow deletion may be a heterozygous deletion, meaning that only one copy of the gene / segment is missing. A deep deletion may be a homozygous deletion, meaning that both copies of the gene / segment are missing.

[0153] The gene fragment described in this disclosure may contain a continuous nucleotide sequence of a reference gene comprising at least 2%, such as at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, or at least 50% of the complete nucleotide sequence of a reference gene.

[0154] Gene amplification and deletion can be evaluated using copy number analysis algorithms, such as Genomic Identification of Significant Targets in Cancer (GISTIC) 2.0, described in Mermel et al., Genome Biology, 12:R41.

[0155] As described in this disclosure, genetic variations may result in increased or decreased expression of a given gene. "Expression" can refer to gene expression and / or protein expression. Gene expression can be assessed by techniques that detect and / or quantify RNA transcribed from the relevant gene and / or RNA encoded by the gene product of the relevant gene. Such techniques include, for example, quantitative real-time PCR (qRT-PCR). Protein expression can be assessed by techniques that detect and / or quantify peptides / peptides encoded by the relevant gene. Such techniques include, for example, antibody-based methods such as Western blotting, immunohistochemistry, immunocytochemistry, flow cytometry, etc.

[0156] It should be noted that protein expression of a given gene can also refer to the expression of the protein encoded by that gene. For example, "protein expression of MET" can also be expressed as "expression of c-Met".

[0157] As described in this disclosure, genetic variations may result in increased or decreased activity of the gene product of a given gene. As used herein, "gene product" refers to the molecule produced by the expression of a given gene. A gene product may be gene-encoded RNA, such as RNA transcribed from the gene, or RNA produced by post-transcriptional processing of RNA transcribed from the gene. A gene product may also be a gene-encoded peptide / polypeptide, such as a peptide / polypeptide formed by translating RNA transcribed from a gene (e.g., after post-transcriptional processing), or a peptide / polypeptide formed by post-translational processing of a protein translated from RNA transcribed from a gene. In some embodiments, the gene product is a gene-encoded peptide / polypeptide.

[0158] The activity of a gene product (such as a gene-encoded peptide / polypeptide) can be any functional property exhibited by that gene product. In some embodiments, the activity of a gene product refers to the functional property exhibited by the gene product encoded by the wild-type allele of the relevant gene. For example, the activity of the MET gene product can be a functional property exhibited by c-Met, such as tyrosine kinase activity.

[0159] Genetic variations that lead to increased expression of a given gene (gene and / or protein) are known or predicted to induce one or more of the following in cells containing such variations: increased gene copy number (e.g., more than two copies); enhanced gene transcription; increased levels of gene-encoded RNA; decreased degradation of gene-encoded RNA; increased levels of gene product; increased levels of gene-encoded peptides / peptides; increased levels of normal splicing of gene-encoded pre-mRNA; enhanced translation of gene-encoded peptides / peptides into mRNA; increased levels of normal post-translational processing of gene-encoded peptides / peptides; increased levels of normal transport of gene-encoded peptides / peptides; and / or decreased degradation of gene-encoded peptides / peptides. Genetic variations that lead to enhanced activity of a given gene product are known or predicted to induce one or more of the following in cells containing such variations: increased levels of one or more functional properties of the gene product; increased levels of one or more functional properties of the gene-encoded peptide / peptide; the gene product acquiring new functional properties; and / or the gene-encoded peptide / peptide exhibiting new functional properties.

[0160] Genetic variations that result in reduced expression of a given gene (gene and / or protein) are known or predicted to induce one or more of the following in cells containing such variations: reduced gene copy number (e.g., less than two copies); weakened gene transcription; decreased levels of gene-encoded RNA; increased degradation of gene-encoded RNA; decreased levels of gene product; decreased levels of gene-encoded peptides / peptides; decreased levels of normal splicing of gene-encoded pre-mRNA; decreased translation of gene-encoded peptides / peptides into mRNA; decreased levels of normal post-translational processing of gene-encoded peptides / peptides; decreased levels of normal transport of gene-encoded peptides / peptides; and / or increased degradation of gene-encoded peptides / peptides. Genetic variations that result in reduced activity of a given gene product are known or predicted to induce one or more of the following in cells containing such variations: decreased levels of one or more functional properties of the gene product; decreased levels of one or more functional properties of the gene-encoded peptide / peptide.

[0161] The most common nucleotide sequence version of a given gene is called its wild-type allele. The nucleotide sequence version of a given gene that contains genetic variation is called its "variant" or "mutated" allele. It is important to understand that the nucleotide sequence of a variant / mutated allele of a given gene differs from that of the wild-type allele.

[0162] In this document, cancer containing cells with a specific characteristic may be simply referred to as cancer with that characteristic. It should be understood that, in the embodiments described herein, cancer containing cells with a specific characteristic may be or may contain one or more tumors containing cells with that characteristic. That is, when a cancer is described as having a specific genetic variation, mutation state, allele, or genotype, it means that the cells of that cancer have the corresponding genetic variation, mutation state, allele, or genotype.

[0163] For example, describing a cancer as containing a given mutation means that the cancer contains cells with that mutation. Similarly, describing a cancer as homozygous for a given genetic variant / allele means that the cancer contains cells homozygous for that genetic variant / allele. Likewise, describing a cancer as heterozygous for a given genetic variant / allele means that the cancer contains cells heterozygous for that genetic variant / allele.

[0164] When a cancer is described as having a given genetic variant, mutation state, allele, or genotype, one or more cells in the cancer have the corresponding genetic variant, mutation state, allele, or genotype. In some embodiments, if a cancer is described as having a given genetic variant, mutation state, allele, or genotype, then a majority (>50%) of the cells in the cancer have the corresponding genetic variant, mutation state, allele, or genotype. In some embodiments, any one of ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, or 100% of the cells in the cancer have the corresponding genetic variant, mutation state, allele, or genotype. In some implementations, a cancer containing a given genetic variant / mutation / allele / genotype can be a cancer in which >10% (e.g., ≥20%, ≥30%, ≥40%, ≥50%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, or 100%) of the cells contain the genetic variant / mutation / allele / genotype.

[0165] In some implementations, a cancer that does not contain (i.e. lacks) a given genetic variant / mutation / allele / genotype is defined as a cancer in which <25% (e.g., ≤20%, ≤15%, ≤10%, ≤5%, ≤1%, or zero) of the cells contain the genetic variant / mutation / allele / genotype.

[0166] In this document, when describing a cell as containing a given genetic variant / mutation, it can be understood that one or more alleles of the relevant gene contain such a genetic variant / mutation. In some embodiments, a cell containing a given genetic variant / mutation may be a heterozygote containing the allele of said genetic variant / mutation; in some embodiments, a cell containing a given genetic variant / mutation may be a homozygote containing the allele of said genetic variant / mutation.

[0167] When describing a cell as not containing a given genetic variation / mutation, it can be understood that all alleles of the relevant gene do not contain the genetic variation / mutation (i.e., the cell is not homozygous or heterozygous for the mutation / mutation allele).

[0168] The various aspects and embodiments described in this disclosure involve genetic variations that lead to increased MET expression or enhanced activity of gene products.

[0169] Genetic variations known or predicted to increase MET / c-Met expression and / or activity are described in, for example, Tovar and Graveel, Vol. 5, No. 10, 2017, p. 205, which is incorporated herein by reference in its entirety.

[0170] In some implementations, genetic variations that lead to increased MET expression or enhanced activity of the gene product are called MET activating mutations. MET activating mutations include E34 (e.g., E34K), H150 (e.g., H150Y), E168 (e.g., E168D), L269 (e.g., L269V), L299 (e.g., L299F), S323 (e.g., S323G), M362 (e.g., M362T), N375 (e.g., N375S), C358 (e.g., C358Y), R970 (e.g., R970C), R988 (e.g., R988C), P1009 (e.g., P1009S), T1010 (e.g., T1010I), S1058 (e.g., S1058P), exon 14 skipping mutations, A1108 (e.g., A1108S), V1110 (e.g., V1110I), and H1112 (e.g., H1112R, H1112L, H1112I). H1124 (e.g., H1124D), G1137 (e.g., G1137V), M1149 (e.g., M1149T), T1191 (e.g., T1191I), V1206 (e.g., V1206L), L1213 (e.g., L1213V), D1228 (e.g., D1228V), Y1230 (e.g., Y1230C, Y1230H, Y123) Mutations in 0D), Y1235 (e.g., Y1235D), V1238 (e.g., V1238I), D1246 (e.g., D1246N), Y1248 (e.g., Y1248C, Y1248D, Y1248H), K1262 (e.g., K1262R), M1268 (e.g., M1268T, M1268I), and V1312 (e.g., V1312I).

[0171] Therefore, in some implementations, the MET activating mutation is selected from the following mutations: E34 (e.g., E34K), H150 (e.g., H150Y), E168 (e.g., E168D), L269 (e.g., L269V), L299 (e.g., L299F), S323 (e.g., S323G), M362 (e.g., M362T), N375 (e.g., N375S), C358 (e.g., C358Y), R970 (e.g., R970C), R988 (e.g., R988C), P1009 (e.g., P1009S), T1010 (e.g., T1010I), S1058 (e.g., S1058P), exon 14 skipping mutation, A1108 (e.g., A1108S), V1110 (e.g., V1110I), H1112 (e.g., H1112R, H111...). 2L, H1112I), H1124 (e.g., H1124D), G1137 (e.g., G1137V), M1149 (e.g., M1149T), T1191 (e.g., T1191I), V1206 (e.g., V1206L), L1213 (e.g., L1213V), D1228 (e.g., D1228V), Y1230 (e.g., Y1230C, Y1230) H, Y1230D), Y1235 (e.g., Y1235D), V1238 (e.g., V1238I), D1246 (e.g., D1246N), Y1248 (e.g., Y1248C, Y1248D, Y1248H), K1262 (e.g., K1262R), M1268 (e.g., M1268T, M1268I) or V1312 (e.g., V1312I).

[0172] In some implementations, the genetic variant leading to increased MET expression or enhanced gene product activity is or includes MET gene amplification. MET amplification can be identified using techniques well-known in the art, such as in situ hybridization. For example, MET amplification can be assessed using fluorescence in situ hybridization (FISH), see Awad et al., *J. Clin. Oncol.*, 2016, Vol. 34, No. 7, pp. 721-730, or Yeung et al., *J. Thorac. Oncol.*, 2015, No. 10, pp. 1292-300. MET amplification can also be assessed using next-generation sequencing (NGS), see Schubart et al., *Cancers (Basel)*, 2021, Vol. 13, No. 19, p. 5023.

[0173] In some embodiments, cells exhibiting MET amplification contain >2 copies of the MET gene, such as any one or more copies of 3, 4, 5, 6, 7, 8, 9, or 10, or >2 copies of MET gene fragments, such as any one or more copies of 3, 4, 5, 6, 7, 8, 9, or 10. MET amplification can be a result of gene duplication or polychromaticity. In some embodiments, MET-amplified cancer may contain a MET gene to chromosome 7 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, MET-amplified cancer may contain a MET gene to chromosome 7 centromere (CEN7) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cells with MET-amplified cancer may contain a MET gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (e.g., as determined by NGS).

[0174] In some embodiments, the genetic variant leading to increased MET expression or enhanced gene product activity is not or does not contain a MET gene mutation. In some embodiments, the genetic variant leading to increased MET expression or enhanced gene product activity refers to a genetic variant other than a MET mutation (if it is or contains MET amplification, as described above).

[0175] The various aspects and embodiments described in this disclosure also involve genetic variations that lead to increased KRAS expression or enhanced activity of gene products.

[0176] Genetic variants known or predicted to increase KRAS / K-Ras expression and / or activity are described in Hobbs and Der *Cancer Discovery*, Vol. 9, No. 6, pp. 696-698, 2019, which is incorporated herein by reference in its entirety. KRAS activating mutations include G12 mutations (e.g., G12A, G12D, G12R, G12C, G12S, and G12V), G13 mutations (e.g., G13D, G13C), Q61 mutations (e.g., Q61H, Q61L, Q61K, Q61R), A146 mutations (e.g., A146T, A146V), and K117 mutations (e.g., K117N). In some embodiments, the KRAS activating mutation described herein is G12C.

[0177] In some implementations, the genetic variant leading to increased KRAS expression or enhanced gene product activity is or includes KRAS gene amplification. KRAS amplification can be identified using techniques well-known in the art, such as in situ hybridization and NGS. For example, fluorescence in situ hybridization can be used to assess KRAS amplification; see Valtorta et al., *International Journal of Cancer*, 2013, Vol. 133, No. 5, pp. 1259-65.

[0178] In some embodiments, cells containing KRAS amplification contain >2 copies of the KRAS gene, such as any one or more copies of KRAS gene segments from the sequence 3, 4, 5, 6, 7, 8, 9, and 10. KRAS amplification can be a result of gene duplication or polychromatic amplification. In some embodiments, KRAS-amplified cancer may contain a KRAS gene to chromosome 12 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, KRAS-amplified cancer may contain a KRAS gene to chromosome 12 centromere (CEN12) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some implementations, cells of KRAS amplified carcinoma may contain KRAS gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9 or ≥10 (as determined by NGS).

[0179] In some embodiments, the genetic variant leading to increased KRAS expression or enhanced gene product activity is not or does not contain KRAS gene amplification. In some embodiments, the genetic variant leading to increased KRAS expression or enhanced gene product activity refers to genetic variants other than KRAS amplification (such as those containing KRAS activating mutations, as described above).

[0180] The various aspects and embodiments described in this disclosure involve genetic variations that lead to increased PIK3CA expression or enhanced activity of gene products.

[0181] Genetic variants known or predicted to increase PIK3CA / PI3K expression and / or activity are described in Ligresti et al., *Cell Cycle*, Vol. 8, No. 9, 2009, pp. 1352-1358, which is incorporated herein by reference in its entirety. PIK3CA activating mutations include H1047 mutations (e.g., H1047R, H1047L), E542 mutations (e.g., E542K, E542Q), E545 mutations (e.g., E545K), P449 mutations (e.g., P449T), and Q546 mutations (e.g., Q546R). In some embodiments, the PIK3CA activating mutations described herein are Q546R or P449T.

[0182] In some implementations, the genetic variant leading to increased PIK3CA expression or enhanced gene product activity is or includes PIK3CA gene amplification. PIK3CA amplification can be identified using techniques well-known in the art, such as in situ hybridization and NGS. For example, fluorescence in situ hybridization can be used to assess PIK3CA amplification; see Holst et al., ClinCancer Res., 2019, Vol. 25, No. 1, pp. 334-45.

[0183] In some embodiments, cells containing PIK3CA amplification contain >2 copies of the PIK3CA gene, such as any one or more copies of PIK3CA gene segments from 3, 4, 5, 6, 7, 8, 9, and 10. PIK3CA amplification can be a result of gene duplication or polychromaticity. In some embodiments, PIK3CA amplified cancer may contain a PIK3CA gene to chromosome 3 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, PIK3CA amplified cancer may contain a PIK3CA gene to chromosome 3 centromere (CEN3) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some implementations, cells of PIK3CA amplified carcinoma may contain PIK3CA gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9 or ≥10 (e.g., as determined by NGS).

[0184] In some embodiments, the genetic variant leading to increased PIK3CA expression or enhanced gene product activity is not or does not contain PIK3CA gene amplification. In some embodiments, the genetic variant leading to increased PIK3CA expression or enhanced gene product activity refers to genetic variants other than PIK3CA amplification (such as those containing PIK3CA activating mutations, as described above).

[0185] The various aspects and embodiments described in this disclosure involve genetic variations that lead to increased BRAF expression or enhanced activity of gene products.

[0186] Genetic variants known or predicted to increase BRAF / B-Raf expression and / or activity are described in Van Cutsem et al., *Journal of Clinical Immunology*, 2011, Vol. 29, No. 15, pp. 2011-2019, which is incorporated herein by reference in its entirety. BRAF activating mutations include V600 mutations (e.g., V600E, V600K), T119 mutations (e.g., T119S), and L597 mutations (e.g., L597R). In some embodiments, the activating mutations described in this disclosure are V600E, V600K, T119S, or L597R.

[0187] In some implementations, the genetic variation leading to increased BRAF expression or enhanced activity of the gene product is or includes BRAF gene amplification. BRAF amplification can be identified using techniques well-known in the art, such as in situ hybridization and NGS. For example, BRAF amplification can be assessed using fluorescence in situ hybridization; see Corcoran et al., *Science Signal*, 2010, Vol. 3, No. 149, ra84.

[0188] In some embodiments, cells containing BRAF amplification contain >2, such as any one or more copies of the BRAF gene from the sequence 3, 4, 5, 6, 7, 8, 9, and 10, or >2, such as any one or more copies of BRAF gene fragments from the sequence 3, 4, 5, 6, 7, 8, 9, and 10. BRAF amplification can be the result of gene duplication or polychromatic amplification. In some embodiments, BRAF-amplified cancer may contain a BRAF gene to chromosome 7 ratio ≥1 (e.g., determined by in situ hybridization), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, BRAF-amplified cancer may contain a BRAF gene to chromosome 7 centromere (CEN7) ratio ≥1 (e.g., determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cells containing BRAF-amplified cancer may contain a BRAF gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (e.g., determined by NGS).

[0189] In some embodiments, the genetic variant leading to increased BRAF expression or enhanced gene product activity is not or does not contain BRAF gene amplification. In some embodiments, the genetic variant leading to increased BRAF expression or enhanced gene product activity refers to genetic variants other than BRAF amplification (such as those containing BRAF activating mutations, as described above).

[0190] The various aspects and embodiments described in this disclosure involve genetic variations that lead to reduced PTEN expression or decreased activity of gene products.

[0191] Genetic variations known or predicted to reduce PTEN / PTEN expression and / or activity are described in Chang et al., *Biomolecules*, Vol. 9, No. 11, 2019, p. 713, which is incorporated herein by reference in its entirety. PTEN inactivation mutations include R130, R173, R233, K267, and N323 mutations.

[0192] In some implementations, the genetic variant leading to reduced PTEN expression or decreased activity of the gene product is or includes a PTEN gene deletion. PTEN deletions can be identified using techniques well-known in the art, such as in situ hybridization and NGS. For example, PTEN deletions can be assessed using fluorescence in situ hybridization; see Wang et al., *Neoplasia*, 2018, Vol. 20, No. 6, pp. 574-593.

[0193] In some embodiments, cells containing PTEN deletion contain <2, such as 1 or 0, PTEN gene copies. In some embodiments, cancers containing PTEN deletion may have a PTEN gene to chromosome 17 ratio ≤1 (e.g., as determined by ISH). In some embodiments, cancers containing PTEN deletion may have a PTEN gene to chromosome 17 centromere (CEN17) ratio ≤1 (e.g., as determined by ISH). In some embodiments, cells containing PTEN deletion cancer may have a PTEN gene copy number (GCN) <2, such as ≤1.5, ≤1, or ≤0.5 (e.g., as determined by NGS).

[0194] In some embodiments, the genetic variant that leads to reduced PTEN expression or decreased gene product activity is not or does not contain a PTEN mutation. In some embodiments, the genetic variant that leads to reduced PTEN expression or decreased gene product activity refers to a genetic variant other than a PTEN mutation (such as or containing a PTEN deletion, as described above).

[0195] The various aspects and embodiments described in this disclosure relate to genetic variations that lead to increased EGFR expression or enhanced activity of gene products.

[0196] Genetic variants known or predicted to increase EGFR / EGFR expression and / or activity are described in Gazdar, Oncogene, 2009, Vol. 28 (Supplement 1): S24-S31, which is incorporated herein by reference in its entirety. EGFR activating mutations include L858 mutations (e.g., L858R), G719 mutations (e.g., G719S, G719A, G719C), T790 mutations (e.g., T790M), V765 mutations (e.g., V765A), T783 mutations (e.g., T783A), and exon 19 deletion mutations (e.g., deletions involving L747 to E749).

[0197] In some implementations, the genetic variant leading to increased EGFR expression or enhanced activity of the gene product is or includes EGFR gene amplification. EGFR amplification can be identified using techniques well-known in the art, such as in situ hybridization and NGS. For example, EGFR amplification can be assessed using fluorescence in situ hybridization; see French et al., Neuro-Oncology, 2019, Vol. 21, No. 10, pp. 1263-1272.

[0198] In some embodiments, cells containing EGFR amplification contain >2, such as any one or more copies of the EGFR gene from the sequence 3, 4, 5, 6, 7, 8, 9, and 10, or >2, such as any one or more copies of EGFR gene fragments from the sequence 3, 4, 5, 6, 7, 8, 9, and 10. EGFR amplification can be the result of gene duplication or polychromatic amplification. In some embodiments, EGFR-amplified cancer may contain an EGFR gene to chromosome 7 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, EGFR-amplified cancer may contain an EGFR gene to chromosome 7 centromere (CEN7) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cells of EGFR-amplified cancer may contain an EGFR gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (e.g., as determined by NGS).

[0199] The various aspects and embodiments described in this disclosure involve genetic variations that lead to increased TP63 expression or enhanced activity of gene products.

[0200] In some implementations, the genetic variation leading to increased TP63 expression or enhanced gene product activity is or includes TP63 gene amplification. TP63 amplification can be identified using techniques well known in the art, such as in situ hybridization and NGS. For example, TP63 amplification can be assessed using fluorescence in situ hybridization; see Massion et al., *Cancer Research*, 2003, Vol. 63, No. 21, pp. 7113-21.

[0201] In some embodiments, cells containing TP63 amplification contain >2 copies of the TP63 gene, such as any one or more copies of TP63 gene segments from 3, 4, 5, 6, 7, 8, 9, and 10. TP63 amplification can be a result of gene duplication or polychromaticity. In some embodiments, TP63-amplified cancer may contain a TP63 gene to chromosome 3 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, TP63-amplified cancer may contain a TP63 gene to chromosome 3 centromere (CEN3) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some implementations, TP63 amplified cancer cells may contain TP63 gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9 or ≥10 (e.g., as determined by NGS).

[0202] The various aspects and embodiments described in this disclosure involve genetic variations that lead to increased SOX2 expression or enhanced activity of gene products.

[0203] In some implementations, the genetic variation leading to increased SOX2 expression or enhanced activity of the gene product is or includes SOX2 gene amplification. SOX2 amplification can be identified using techniques well known in the art, such as in situ hybridization and NGS. For example, SOX2 amplification can be assessed using fluorescence in situ hybridization; see Wilbertz et al., Modern Pathology, 2011, Vol. 24, pp. 944-953.

[0204] In some embodiments, cells containing SOX2 amplification contain >2 copies of the SOX2 gene, such as any one or more copies of SOX2 gene fragments from 3, 4, 5, 6, 7, 8, 9, and 10. SOX2 amplification can be a result of gene duplication or polychromatic amplification. In some embodiments, SOX2-amplified cancer may contain a SOX2 gene to chromosome 3 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, SOX2-amplified cancer may contain a SOX2 gene to chromosome 3 centromere (CEN3) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some implementations, SOX2 amplified cancer cells may contain SOX2 gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9 or ≥10 (e.g., as determined by NGS).

[0205] The various aspects and embodiments described in this disclosure relate to genetic variations that result in increased expression of one or more genes located on the long arm of chromosome 3 or enhanced activity of their gene products. In some embodiments, the genetic variation resulting in increased expression of one or more genes located on the long arm of chromosome 3 or enhanced activity of their gene products is or includes an amplification of the relevant gene. Amplification of one or more genes located on the long arm of chromosome 3 can be identified by techniques well known in the art, such as in situ hybridization and NGS.

[0206] In some embodiments, cells containing amplifications of one or more genes located on the long arm of chromosome 3 contain >2 copies of any one or more of the relevant genes, such as those from chromosomes 3, 4, 5, 6, 7, 8, 9, and 10, or >2 copies of fragments of any one or more of the relevant genes from chromosomes 3, 4, 5, 6, 7, 8, 9, and 10. Amplifications of one or more genes located on the long arm of chromosome 3 can be a result of gene duplication or polychromatosis. In some embodiments, cancers containing amplifications of one or more genes located on the long arm of chromosome 3 may contain a ratio of the relevant gene to chromosome 3 ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cancers containing amplifications of one or more genes located on the long arm of chromosome 3 may contain a ratio of the relevant gene to the centromere (CEN3) of chromosome 3 ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some implementations, cancer cells containing amplifications of one or more genes located on the long arm of chromosome 3 may contain relevant gene copy numbers (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (e.g., as determined by NGS).

[0207] As described in the various aspects and embodiments of this disclosure, genetic variations that result in increased expression of one or more genes located in the range of 3q26 to 3q28 (e.g., 3q26, 3q27, or 3q28) or enhanced activity of their gene products are involved. In some embodiments, the genetic variation resulting in increased expression of one or more genes located in the range of 3q26 to 3q28 (e.g., 3q26, 3q27, or 3q28) or enhanced activity of their gene products is or includes an amplification of the relevant gene. Amplification of one or more genes located in the range of 3q26 to 3q28 (e.g., 3q26, 3q27, or 3q28) can be identified by techniques well known in the art, such as in situ hybridization and NGS.

[0208] In some embodiments, cells containing amplifications of one or more genes located within 3q26 to 3q28 (e.g., within 3q26, 3q27, or 3q28) contain >2 copies of any one or more of the relevant gene segments, such as 3, 4, 5, 6, 7, 8, 9, or 10. Amplifications of one or more genes located within 3q26 to 3q28 (e.g., within 3q26, 3q27, or 3q28) can be a result of gene duplication or polychromaticity. In some embodiments, cancers containing amplifications of one or more genes located within 3q26 to 3q28 (e.g., within 3q26, 3q27, or 3q28) may contain a relevant gene to chromosome 3 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cancers containing amplifications of one or more genes located in 3q26 to 3q28 may have a relevant gene to chromosome 3 centromere (CEN3) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cancer cells containing amplifications of one or more genes located in 3q26 to 3q28 (e.g., within 3q26, 3q27, or 3q28) may have a relevant gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (e.g., as determined by NGS).

[0209] Genes located between 3q26 and 3q28 include TP63, PIK3CA, and SOX2. Genes located at 3q26 include PIK3CA (3q26.32) and SOX2 (3q26.33). Genes located at 3q28 include TP63.

[0210] The various aspects and embodiments described in this disclosure involve genetic variations that lead to amplification of the long arm of chromosome 3. Amplification of the long arm of chromosome 3 may be the result of multiple chromosomes.

[0211] Genes located on the long arm of chromosome 3 include: ADIPOQ, AMOTL2, ARHGAP31, TIMMDC1, C3orf70, CAMPD1, CCDC80, CD200R1, CHST13, CLDND1, CPN2, CPOX, DPPA2, DTX3L, DZIP3, EAF2, EFCC1, ETM1, ETV5, FAM3D, FAM43A, FAM162A, FBXO40, FILIP1L, GYG1, HACD2, HGD, IFT122, KIAA1257, LINC01279, LNCR5, LM. LN, LRRC15, LSG1, MB21D2, MCCC1, MORC1, MYLK, NEPRO, NFKBIZ, OTOL1, PARP14, PCCB, PDCD10, PIK3CA, PISRT1, PROSER1, RAB7, RASA2, RETNLB, RHO, RIOX2, SELT, SENP7, SERP1, SOX2, SOX2OT, SPG14, SRPRB, TEX55, TMEM44, TM4SF1, TMPRSS7, TP63, TRAT1, USH3A, ZBED2, and ZNF9. In some embodiments, one or more genes located on the long arm of chromosome 3 are selected from TP63, PIK3CA, and SOX2.

[0212] The various aspects and embodiments described in this disclosure relate to genetic variations leading to 3q26 amplification. The various aspects and embodiments described in this disclosure relate to genetic variations leading to 3q27 amplification. The various aspects and embodiments described in this disclosure relate to genetic variations leading to 3q28 amplification. The various aspects and embodiments described in this disclosure relate to genetic variations leading to amplification from 3q26 to 3q28. Such amplification may be the result of multiple chromosomes.

[0213] The various aspects and embodiments described in this disclosure relate to genetic variations that result in increased expression of one or more genes located on the short arm of chromosome 7 or enhanced activity of their gene products. In some embodiments, the genetic variation resulting in increased expression of one or more genes located on the short arm of chromosome 7 or enhanced activity of their gene products is or includes an amplification of the relevant gene. Amplification of one or more genes located on the short arm of chromosome 7 can be identified by techniques well known in the art, such as in situ hybridization and NGS.

[0214] In some embodiments, cells containing amplifications of one or more genes located on the short arm of chromosome 7 contain copies of >2, such as any one or more of the relevant genes from 3, 4, 5, 6, 7, 8, 9, and 10, or copies of >2, such as any one or more fragments of the relevant genes from 3, 4, 5, 6, 7, 8, 9, and 10. Amplifications of one or more genes located on the short arm of chromosome 7 can be a result of gene duplication or polychromatosis. In some embodiments, cancers containing amplifications of one or more genes located on the short arm of chromosome 7 may contain a relevant gene to chromosome 7 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, cancers containing amplifications of one or more genes located on the short arm of chromosome 7 may contain a relevant gene to chromosome 7 centromere (CEN7) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some implementations, cancer cells containing amplifications of one or more genes located on the short arm of chromosome 7 may contain relevant gene copy numbers (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (as determined by NGS).

[0215] The various aspects and embodiments described in this disclosure relate to genetic variations that result in increased expression of one or more genes located at 7p11 or enhanced activity of their gene products. In some embodiments, the genetic variation resulting in increased expression of one or more genes located at 7p11 or enhanced activity of their gene products is or includes an amplification of the relevant gene. Amplification of one or more genes located at 7p11 can be identified using techniques well known in the art, such as in situ hybridization and NGS.

[0216] In some embodiments, cells containing amplification of one or more genes located at 7p11 contain copies of >2, such as any one or more copies of the relevant gene from 3, 4, 5, 6, 7, 8, 9, 10, or >2, such as copies of any one or more fragments of the relevant gene from 3, 4, 5, 6, 7, 8, 9, 10. Amplification of one or more genes located at 7p11 can be a result of gene duplication or polychromaticity. In some embodiments, cancers containing amplification of one or more genes located at 7p11 may contain a ratio of the relevant gene to chromosome 7 ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, ≥2.5. In some embodiments, cancers containing amplification of one or more genes located at 7p11 may contain a ratio of the relevant gene to the centromere of chromosome 7 (CEN7) ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, ≥2.5. In some implementations, cancer cells containing amplification of one or more genes located at 7p11 may contain a related gene copy number (GCN) >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (as determined by NGS).

[0217] Genes located at 7p11 include EGFR (7p11.2).

[0218] The various aspects and embodiments described in this disclosure relate to genetic variations that lead to amplification of the short arm of chromosome 7. The various aspects and embodiments described in this disclosure also relate to genetic variations that lead to amplification of 7p11. Such amplification may be the result of multiple chromosomes.

[0219] The various aspects and embodiments described in this disclosure relate to genetic variations that result in reduced expression of one or more genes located on the short arm of chromosome 3 or decreased activity of their gene products. In some embodiments, the genetic variation resulting in reduced expression of one or more genes located on the short arm of chromosome 3 or decreased activity of their gene products is or includes the loss or deletion of the relevant gene. The loss or deletion of one or more genes located on the short arm of chromosome 3 can be identified by techniques well known in the art, such as in situ hybridization and NGS.

[0220] In some embodiments, cells containing deletions of one or more genes located on the short arm of chromosome 3 contain <2, such as 1 or 0, copies of the relevant gene, or <2, such as 1 or 0, copies of the relevant gene fragment. In some embodiments, cancers containing deletions of one or more genes located on the short arm of chromosome 3 may contain a relevant gene to chromosome 3 copy number ≤1 (e.g., as determined by ISH). In some embodiments, cancers containing deletions of one or more genes located on the short arm of chromosome 3 may contain a relevant gene to chromosome 3 centromere (CEN3) ratio ≤1 (e.g., as determined by ISH). In some embodiments, cells containing deletions of one or more genes located on the short arm of chromosome 3 may contain a relevant gene copy number (GCN) <2, such as ≤1.5, ≤1, or ≤0.5 (e.g., as determined by NGS).

[0221] The various aspects and embodiments described in this disclosure relate to genetic variations that result in reduced expression of one or more genes located at 3p21 (e.g., 3p21.3) or decreased activity of their gene products. In some embodiments, the genetic variation resulting in reduced expression of one or more genes located at 3p21 (e.g., 3p21.3) or decreased activity of their gene products is or includes the deletion of the relevant gene. Deletions of one or more genes located at 3p21 (e.g., 3p21.3) can be identified using techniques well known in the art, such as in situ hybridization and NGS.

[0222] In some embodiments, cells containing deletions of one or more genes located at 3p21 (e.g., 3p21.3) contain <2, such as 1 or 0, copies of the relevant gene, or <2, such as 1 or 0, copies of the relevant gene fragment. In some embodiments, cancers containing deletions of one or more genes located at 3p21 (e.g., 3p21.3) may have a relevant gene to chromosome 3 copy number ≤1 (e.g., as determined by ISH). In some embodiments, cancers containing deletions of one or more genes located at 3p21 may have a relevant gene to chromosome 3 centromere (CEN3) ratio ≤1 (e.g., as determined by ISH). In some embodiments, cells containing deletions of one or more genes located at 3p21 (e.g., 3p21.3) may have a relevant gene copy number (GCN) <2, such as ≤1.5, ≤1, or ≤0.5 (e.g., as determined by NGS).

[0223] Genes located at 3p21 (e.g., 3p21.3) include TUSC2.

[0224] The various aspects and embodiments described in this disclosure relate to genetic variations that result in deletions of the short arm of chromosome 3. The various aspects and embodiments described in this disclosure relate to genetic variations that result in deletions of 3p21.

[0225] This disclosure relates to genetic variations in genes encoding factors related to HER3-mediated signal transduction.

[0226] The various aspects and embodiments described in this disclosure relate to genetic variations that lead to increased expression of positive regulators of HER3-mediated signaling or enhanced activity of their gene products. The various aspects and embodiments described in this disclosure relate to genetic variations that lead to increased expression of positive regulators of the MAPK / ERK signaling pathway or enhanced activity of their gene products. The various aspects and embodiments described in this disclosure relate to genetic variations that lead to increased expression of positive regulators of the PI3K / AKT / mTOR signaling pathway or enhanced activity of their gene products.

[0227] The various aspects and embodiments described in this disclosure relate to genetic variations that result in reduced expression of negative regulators of HER3-mediated signaling or decreased activity of their gene products. The various aspects and embodiments described in this disclosure relate to genetic variations that result in reduced expression of negative regulators of the MAPK / ERK signaling pathway or decreased activity of their gene products. The various aspects and embodiments described in this disclosure relate to genetic variations that result in reduced expression of negative regulators of the PI3K / AKT / mTOR signaling pathway or decreased activity of their gene products.

[0228] HER3-mediated signaling can be analyzed by detecting HER3-mediated signaling-related phenomena, such as cell proliferation, and / or phosphorylation of one or more signaling molecules in the PI3K / AKT / mTOR and MAPK / ERK signaling pathways. For example, PI3K / AKT / mTOR and / or MAPK / ERK signaling levels can be assessed by detecting and quantifying phosphorylation of one or more components in the PI3K / AKT / mTOR and / or MAPK / ERK pathways. Such analyses can be performed in vitro in cell-based assays of HER3-mediated signaling, as described in Example 4.3 of WO2019 / 185878A1, 8.9.

[0229] As used in this paper, a "positive regulator" of signal transduction in a given pathway (such as a positive regulator of HER3-mediated signal transduction, a positive regulator of MAPK / ERK pathway signal transduction, or a positive regulator of PI3K / AKT / mTOR pathway signal transduction) refers to a factor whose expression / activity typically has a positive effect (i.e., enhances or amplifies) on the signal transduction of the relevant pathway. Increased expression levels and / or activity of positive regulators may lead to enhanced signal transduction in the relevant pathway (as determined by analysis of such signal-related phenomena); decreased expression levels and / or activity of positive regulators may lead to weakened signal transduction in the relevant pathway.

[0230] A "negative regulator" of signal transduction in a given pathway (such as a negative regulator of HER3-mediated signal transduction, a negative regulator of MAPK / ERK signal transduction, or a negative regulator of PI3K / AKT / mTOR signal transduction) refers to a factor whose expression / activity typically has a negative effect (i.e., inhibition or antagonism) on the signal transduction of the relevant pathway. Increased expression levels and / or activity of negative regulators may lead to decreased signal transduction levels in the relevant pathway (as determined by analysis of such signal-related phenomena); decreased expression levels and / or activity of negative regulators may lead to increased signal transduction levels in the relevant pathway.

[0231] In some embodiments, the gene encoding a positive regulator of HER3-mediated signal transduction may be selected from: KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, BRAF, and MET. In some embodiments, the gene encoding a positive regulator of HER3-mediated signal transduction may be selected from: KRAS, PIK3CA, PIK3CB, BRAF, and MET. In some embodiments, the positive regulator encoding HER3-mediated signal transduction is KRAS. In some embodiments, the positive regulator encoding HER3-mediated signal transduction is PIK3CA. In some embodiments, the positive regulator encoding HER3-mediated signal transduction is PIK3CB. In some embodiments, the positive regulator encoding HER3-mediated signal transduction is BRAF. In some embodiments, the positive regulator encoding HER3-mediated signal transduction is MET.

[0232] In some implementations, the gene encoding the positive regulator of HER3-mediated signal transduction may be selected from: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5.

[0233] In some embodiments, the gene encoding the positive regulator of MAPK / ERK pathway signaling is selected from: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, and BRAF. In some embodiments, the gene encoding the positive regulator of MAPK / ERK pathway signaling is KRAS. In some embodiments, the gene encoding the positive regulator of MAPK / ERK pathway signaling is BRAF.

[0234] In some implementations, the genes encoding positive regulators of signal transduction in the MAPK / ERK pathway are selected from: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1.

[0235] In some embodiments, the gene encoding the positive regulator of signal transduction in the PI3K / AKT / mTOR pathway may be selected from: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CA, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the gene encoding the positive regulator of signal transduction in the PI3K / AKT / mTOR pathway is PIK3CA. In some embodiments, the gene encoding the positive regulator of signal transduction in the PI3K / AKT / mTOR pathway is PIK3CB.

[0236] In some implementations, the genes encoding the positive regulators of signal transduction in the PI3K / AKT / mTOR pathway are selected from: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5.

[0237] In some embodiments, the gene encoding a negative regulator of HER3-mediated signal transduction may be selected from: PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the gene encoding a negative regulator of PI3K / AKT / mTOR signal transduction may be selected from: PTEN, PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1. In some embodiments, the gene encoding a negative regulator of MAPK / ERK signal transduction may be NF1. In some embodiments, the gene encoding a negative regulator of HER3-mediated signal transduction is PTEN. In some embodiments, the gene encoding a negative regulator of PI3K / AKT / mTOR signal transduction is PTEN.

[0238] In some embodiments, the gene encoding the negative regulator of HER3-mediated signal transduction may be selected from: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the gene encoding the negative regulator of PI3K / AKT / mTOR pathway signal transduction may be selected from: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1.

[0239] In some embodiments, the cancer to be treated / prevented comprises cells expressing HER3 ligands (such as NRG1 and / or NRG2). In some embodiments, the cancer to be treated / prevented comprises cells whose NRG1 and / or NRG2 expression levels are higher than those in corresponding non-cancerous / non-tumor tissues. Such cancers can be described as comprising cells that overexpress NRG1 and / or NRG2.

[0240] The HER3-binding antigen-binding molecules described herein bind HER3 with extremely high affinity when HER3 binds to NRG (when HER3 is in the "open" conformation) and when HER3 does not bind to NRG (when HER3 is in the "closed" conformation). Therefore, they are particularly suitable for the treatment / prevention of cancers characterized by HER3 ligand expression / overexpression, such as cancers or tumors containing cells that express / overexpress HER3 ligands.

[0241] The various aspects and embodiments described in this disclosure involve genetic variations (such as mutations or gene amplifications) that lead to increased expression of HER3 ligands (genes and / or proteins).

[0242] Genetic variations that lead to increased HER3 ligand expression can result in the expression of the HER3 ligand gene or protein, while in equivalent cells without such variations, the genomic nucleic acids do not express and / or encode the HER3 ligand. That is, the HER3 ligand may be a neoantigen resulting from genetic variations, so "increased expression" may originate from a lack of expression. For example, cells containing a CD74-NRG1 gene fusion will show increased expression of the CD74-NRG1 fusion peptide encoded by the gene fusion, compared to cells lacking the CD74-NRG1 gene fusion.

[0243] Genetic variations that lead to increased HER3 ligand expression may result in increased expression of the HER3 ligand gene or protein relative to the ligand encoded by genomic nucleic acids in equivalent cells that do not contain the variation. For example, cells containing genetic variations may lead to increased transcriptional levels of nucleic acids encoding NRG1 relative to the transcriptional levels of NRG1-encoding nucleic acids in equivalent cells that do not contain the variation.

[0244] In some embodiments, a genetic variation that leads to increased HER3 ligand expression may result in an increased gene expression level of that HER3 ligand relative to an equivalent cell that does not contain the genetic variation. In some embodiments, a genetic variation that leads to increased HER3 ligand expression may result in an increased HER3 ligand protein expression level relative to an equivalent cell that does not contain the genetic variation.

[0245] In some embodiments, a genetic variation that leads to increased HER3 ligand expression, relative to an equivalent cell that does not contain the genetic variation, can result in elevated levels of HER3 ligand on or within the cell surface of the cell containing the genetic variation. In some embodiments, a genetic variation that leads to increased HER3 ligand expression, relative to an equivalent cell that does not contain the genetic variation, can result in elevated secretion levels of HER3 ligand in the cell containing the genetic variation.

[0246] Cells exhibiting higher HER3 ligand expression levels compared to reference cells (e.g., due to mutation) can be described as "overexpressing" or "upregulating" HER3 ligand expression. For example, a cancer containing cells carrying a genetic variant that increase HER3 ligand expression compared to equivalent cells lacking the genetic variant can be termed a cancer containing cells with HER3 ligand overexpression / upregulation. In some implementations, the reference cells lacking the genetic variant can be non-cancerous cells (e.g., equivalent cell types) or cancerous cells (e.g., similar tumors).

[0247] In this document, "HER3 ligand" generally refers to a molecule capable of binding to HER3 through a HER3 ligand-binding region formed by HER3 domains I and III. In some embodiments, the HER3 ligand binds to HER3 by interacting with HER3 domains I and / or III. Exemplary HER3 ligands include neural regulatory proteins such as NRG1 and NRG2, which bind to HER3 by interacting with the HER3 ligand-binding region through their own EGF-like domains.

[0248] The HER3 ligand is preferably capable of binding to and stimulating signal transduction via the HER3 receptor and / or a HER3-containing receptor complex. As is understood in this disclosure, the HER3-containing receptor complex may also contain HER3 interacting partners as described herein, such as HER2, EGFR, HER4, HGFR, IGF1R, and / or cMet.

[0249] In some embodiments, the HER3 ligand can bind to HER3 receptor / receptor complexes expressed in cells other than those with increased HER3 ligand expression. For example, in some embodiments, the HER3 ligand can bind to cancer cells expressing HER3.

[0250] In some implementations, the HER3 ligand can bind to the HER3 receptor / receptor complex expressed by cells with increased HER3 ligand expression.

[0251] In some implementations, the cancer to be treated / prevented comprises: (i) cells expressing HER3, and (ii) the cells expressing HER3 ligands (such as those with increased HER3 ligand expression, as a result of genetic variations that lead to increased HER3 ligand expression).

[0252] In some implementations, the cancer to be treated / prevented comprises cells that are: (i) cells that express HER3, and (ii) cells that also express HER3 ligands (e.g., cells with increased HER3 ligand expression, such as as a result of genetic variations that lead to increased HER3 ligand expression).

[0253] In some embodiments, the HER3 ligand comprises or consists of an amino acid sequence of the HER3 binding domain of the HER3 ligand or a derived amino acid sequence of the HER3 binding domain of the HER3 ligand. The amino acid sequence derived from the HER3 binding domain of the HER3 ligand may contain at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the amino acid sequence from which it is derived.

[0254] In some embodiments, the HER3 ligand comprises an EGF-like domain or a HER3-binding fragment thereof capable of binding HER3. In some embodiments, the HER3-binding EGF-like domain / fragment is or is derived from a member of the EGF family (such as heparin-binding EGF-like growth factor (HB-EGF), transforming growth factor-α (TGF-α), amphotericin (AR), epithelial regulatory protein (EPR), epigenetic protein, β-cytokinin (BTC), NRG1, NRG2, NRG3, or NRG4).

[0255] Exemplary HER3 ligands include neuroregulatory proteins (NRGs). Neuroregulatory proteins include NRG1 (including its α, α2b, and α3 isoforms), NRG2, NRG3, and NRG4. In some embodiments, NRGs are selected from NRG1, NRG2, NRG3, and NRG4. In some embodiments, NRGs are selected from NRG1 and NRG2.

[0256] The EGF-like domain of human NRG1 is composed of amino acids 178-222 of UniProt:Q02297-1, through which it binds to HER3. The EGF-like domain of human NRG2 is composed of amino acids 341-382 of UniProt:O14511-1. The EGF-like domain of human NRG3 is composed of amino acids 286-329 of UniProt:B9EGV5-1. The EGF-like domain of human NRG4 is composed of amino acids 5-46 of UniProt:Q8WWG1-1. In some embodiments, the EGF-like domain / fragment comprises or consists of an amino acid sequence having at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the EGF-like domain of the NRG (NRG1, NRG2, NRG3, or NRG4).

[0257] In some implementations, the HER3 ligand is not an EGFR family protein (such as HER3, HER2, EGFR, HER4, HGFR, IGF1R, cMet).

[0258] In some embodiments, the genetic variant leading to increased HER3 ligand expression is an NRG gene fusion. In some embodiments, the HER3 ligand is the product of an NRG gene fusion (i.e., the encoded polypeptide). In some embodiments, the cancer comprises cells with an NRG gene fusion. As used herein, “NRG gene fusion” refers to a genetic variant encoding a polypeptide comprising: (i) the amino acid sequence of an NRG protein (such as NRG1, NRG2, NRG3, or NRG4; such as NRG1 or NRG2), and (ii) the amino acid sequence of a protein other than an NRG protein.

[0259] NRG gene fusions can be identified and characterized by appropriate molecular assays, methods well known to those skilled in the art.

[0260] The NRG gene fusion described in this disclosure is preferably in the forward orientation (i.e., the nucleotide sequence encoding NRG is located at the 3' end of the transcript) and encodes a fusion polypeptide containing an EGF-like domain capable of binding HER3.

[0261] It should be understood that the NRG gene fusion preferably encodes the HER3 ligand described herein. In some embodiments, the NRG gene fusion encodes a polypeptide comprising the HER3-binding domain of the NRG protein. In some embodiments, the NRG gene fusion encodes a polypeptide comprising the EGF-like domain of the NRG protein, or an amino acid sequence capable of binding to HER3 and having at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the EGF-like domain of the NRG protein.

[0262] In some embodiments, the NRG gene fusion encodes a fusion polypeptide containing a transmembrane domain. In some embodiments, the NRG gene fusion encodes a fusion polypeptide containing a transmembrane domain of a protein other than the NRG protein.

[0263] In some embodiments, the NRG gene fusion is an NRG1 gene fusion. In some embodiments, the NRG1 gene fusion encodes a polypeptide containing the EGF-like domain of NRG1, or an amino acid sequence that can bind to HER3 and has at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the EGF-like domain of NRG1.

[0264] NRG1 gene fusions are operable fusions in tissues of unknown origin that are targeted by HER3 inhibition. These fusions arise from multiple interchromosomal gene translocations, leading to overproduction and aberrant activation of HER3-binding NRG1 ligands, thereby inducing tumorigenesis. Studies have confirmed a causal relationship between HER3 pathway activation and NRG1 gene fusions. NRG1 fusions are enriched in subjects with mucinous non-small cell lung cancer and pancreatic ductal adenocarcinoma: 8–32% of mucinous non-small cell lung cancer subjects have NRG1 gene fusions, while 10–20% (up to 70% in small studies) of subjects with KRAS wild-type PDAC have detectable NRG1 gene fusions. The therapeutic efficacy of monoclonal anti-HER3 antibody therapy targeting NRG1 gene fusions has been demonstrated in WO2021 / 048274A1.

[0265] NRG1 gene fusions are described in WO2021 / 048274A1, WO2018 / 182422A1, WO2019 / 051155A1, Dhanasekaran et al., *Nature Communications*, 2014, Vol. 5, p. 5893; Drilon et al., *Cancer Discovery*, 2018, Vol. 8, No. 6, pp. 686-695; Nagasaka et al., *Journal of Thoracic Oncology*, 2019, Vol. 14, No. 8, pp. 1354-1359; and Jonna et al., *Clinical Cancer Research*, 2019, Vol. 25, No. 16, pp. 4966-4972. All these references are incorporated herein by reference. The diversity of NRG1 gene fusions may stem from NRG1's location on chromosome 8, which is particularly susceptible to genomic translocation events. (See *Genes, Chromosomes, Cancer*, 2003, Vol. 37, No. 4, pp. 333-45).

[0266] In some implementations, the NRG1 gene fusion is selected from: CLU-NRG1, CD74-NRG1, DOC4-NRG1, SLC3A2-NRG1, RBPMS-NRG1, WRN-NRG1, SDC4-NRG1, RAB2IL1-NRG1, VAMP2-NRG1, KIF13B-NRG1, THAP7-NRG1, SMAD4-NRG1, MDK-NRG1, TNC-NRG1, DIP2B-NRG1, MR PL13-NRG1, PARP8-NRG1, ROCK1-NRG1, DPYSL2-NRG1, ATP1B1-NRG1, CDH6-NRG1, APP-NRG1, AKAP13-NRG1, THBS1-NRG1, FOXA1-NRG1, PDE7A-NRG1, RAB3IL1-NRG1, CDK1-NRG1, BMPRIB-NRG1, TNFRSF10B-NRG1, and MCPH1-NRG1. In some embodiments, the NRG1 gene is fused to CLU-NRG1.

[0267] The CD74-NRG1 gene fusion is described in Fernandez-Cuesta et al., *Cancer Discovery*, 2014, Vol. 4, pp. 415-422, and in Nakaoku et al., *Clinical Cancer Research*, 2014, Vol. 20, pp. 3087-3093. The DOC4-NRG1 gene fusion is described in Liu et al., *Oncogene*, 1999, Vol. 18, No. 50, pp. 7110-7114, and in Wang et al., *Oncogene*, 1999, Vol. 18, No. 41, pp. 5718-5721. The SLC3A2-NRG1 gene fusion is described in Nakaoku et al., *Clinical Cancer Research*, 2014, Vol. 20, pp. 3087-3093; Shin et al., *Oncotarget*, 2016, Vol. 7, pp. 69450-69465; and Shin et al., *Molecular Cancer Therapy*, 2018, Vol. 17, No. 9, pp. 2024-2033. The RBPMS-NRG1, WRN-NRG1, RAB2IL1-NRG1, and SDC4-NRG1 gene fusions are described in Dhanasekaran et al., *Nature Communications*, 2014, Vol. 5, p. 5893. VAMP2-NRG1 gene fusion was described by Jung et al., *J Thorac Oncol.*, 2015, Vol. 10, No. 7, pp. 1107-1111, and by Shim et al., *J Thorac Oncol.*, 2015, Vol. 10, No. 8, pp. 1156-1162. KIF13B-NRG1 gene fusion was described by Xia et al., *Int J Surg Pathol.*, 2017, Vol. 25, No. 3, pp. 238-240. SMAD4-NRG1, AKAP13-NRG1, THBS1-NRG1, FOXA1-NRG1, PDE7A-NRG1, RAB3IL1-NRG1, and THAP7-NRG1 gene fusions were described by Drilon et al., *Cancer Discov.*, 2018, Vol. 8, No. 6, pp. 686-695. The gene fusions of MDK-NRG1, TNC-NRG1, DIP2B-NRG1, MRPL13-NRG1, PARP8-NRG1, ROCK1-NRG1, and DPYSL2-NRG1 are described by Jonna et al., Clin Cancer Res., 2019, Vol. 25, No. 16, pp. 4966-4972.The ATP1B1-NRG1 gene fusion was described by Drilon et al., *Cancer Discov.*, 2018, Vol. 8, No. 6, pp. 686-695, and by Jones et al., *Annals of Oncology*, 2017, Vol. 28, pp. 3092-3097. The CLU-NRG1 gene fusion was described by Drilon et al., *Cancer Discov.*, 2018, Vol. 8, No. 6, pp. 686-695, and by Nagasaka et al., *Journal of Thoracic Oncology*, 2019, Vol. 14, No. 8, pp. 1354-1359.

[0268] In some embodiments, the NRG gene fusion is an NRG2 gene fusion. In some embodiments, the NRG2 gene fusion encodes a polypeptide containing the EGF-like domain of NRG2, or an amino acid sequence that can bind HER3 and has at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the EGF-like domain of NRG2.

[0269] NRG2 gene fusions include SLC12A2-NRG2 (see WO2021 / 048274A1, WO2015 / 093557A1) and ZNF208-NRG2 (see Dupain et al., Molecular Therapy, Vol. 27, No. 1, pp. 200-218, 2019).

[0270] cancer

[0271] This disclosure relates to the treatment and prevention of cancer.

[0272] As described in this disclosure, cancer can be any abnormal cell proliferation (or any disease manifested as abnormal cell proliferation), growth, or tumor. The cancer can be benign or malignant. The cancer can be primary or secondary (e.g., metastatic). The growth or tumor can be any abnormal cell growth or proliferation and can be located in any organ / tissue (and / or originate from cells in any organ / tissue).

[0273] Cancer may originate from the following cells: adrenal glands, adrenal medulla, anus, appendix, bladder, blood, bones, bone marrow, brain, breast, cecum, central nervous system (including or excluding the brain), cerebellum, cervix, colon, duodenum, endometrium, epithelial cells (such as renal epithelium), gallbladder, esophagus, glial cells, heart, ileum, jejunum, kidneys, lacrimal glands, larynx, liver, lungs, lymph nodes, lymphoblasts, maxilla, mediastinum, mesentery, myometrium, nasopharynx, greater omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous system, peritoneum, pleura, prostate, salivary glands, sigmoid colon, skin, small intestine, soft tissue, spleen, stomach, testes, thymus, thyroid gland, tongue, tonsils, trachea, uterus, vulva, and / or leukocytes.

[0274] The cancer may be or include one or more tumors. The cancer may be glioma, medulloblastoma, meningioma, neurofibroma, ependymoma, schwannoma, neurofibrosarcoma, astrocytoma and oligodendroglioma, melanoma, mesothelioma, myeloma, lymphoma, non-Hodgkin lymphoma (NHL), Hodgkin lymphoma, cutaneous T-cell lymphoma (CTCL), leukemia, chronic myeloid leukemia (CML), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS), liver cancer, epidermoid carcinoma, prostate cancer, breast cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, ovarian cancer, pancreatic cancer, thymic carcinoma, hematologic malignancy, or sarcoma.

[0275] In some embodiments, the cancers described in this disclosure are selected from: solid tumors, breast cancer, breast carcinoma, ductal carcinoma, gastric cancer, gastric carcinoma, gastric adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, head and neck cancer, squamous cell carcinoma of the head and neck (SCCHN), lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, ovarian cancer, ovarian carcinoma, ovarian serous adenocarcinoma, renal cancer, renal cell carcinoma, clear cell renal carcinoma, renal cell adenocarcinoma, papillary renal carcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic duct adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, esophageal cancer, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, bile duct cancer, uterine cancer, endometrial cancer of the uterine corpus, thyroid cancer. (carcinoma), pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma, and thymoma.

[0276] In some embodiments, the cancer is selected from: cancers containing cells expressing / overexpressing EGFR family members (such as HER3, EGFR, HER2, or HER4), cancers containing cells expressing / overexpressing HER3, cancers containing cells expressing / overexpressing EGFR, cancers containing cells expressing / overexpressing both HER3 and EGFR, cancers containing cells with mutations leading to increased HER3 ligand expression, cancers containing cells with mutations leading to increased EGFR ligand expression, cancers containing NRG gene fusion cells, cancers containing NRG1 gene fusion cells, or cancers containing NRG2 gene fusion cells, solid tumors, hematologic malignancies, squamous cell carcinoma, EGFR-amplified squamous cell carcinoma, breast cancer, breast carcinoma, invasive breast cancer, ductal carcinoma, metastatic breast cancer, triple-negative breast cancer, HER2-positive breast cancer, gastric cancer, gastric carcinoma, gastric adenocarcinoma, gastrointestinal adenocarcinoma, colorectal cancer, metastatic colorectal cancer, colon cancer, colorectal cancer. Colorectal cancer, colorectal cancer, colorectal adenocarcinoma, head and neck cancer, squamous cell carcinoma of the head and neck, lung cancer, non-small cell lung cancer, lung adenocarcinoma, invasive mucinous lung adenocarcinoma, squamous cell carcinoma of the lung, squamous cell carcinoma of the lung, squamous non-small cell lung cancer, advanced squamous non-small cell lung cancer, metastatic squamous non-small cell lung cancer, ovarian cancer. (carcinoma), ovarian serous adenocarcinoma, ovarian serous cystadenocarcinoma, fallopian tube cancer, renal cancer, renal cell carcinoma, clear cell renal carcinoma, renal cell adenocarcinoma, papillary renal cell carcinoma, pancreatic cancer, exocrine gland cancer, pancreatic adenocarcinoma, pancreatic duct adenocarcinoma, advanced pancreatic duct adenocarcinoma, metastatic pancreatic duct adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, oral cancer, oropharyngeal cancer, esophageal cancer, esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, bile duct cancer, gallbladder cancer, uterine cancer, endometrial cancer, uterine corpus endometrial cancer, uterine carcinosarcoma, thyroid cancer, thyroid cancer Pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, retinoblastoma, sarcoma, soft tissue sarcoma, peritoneal cancer, thymoma, neuroendocrine tumor, nasopharyngeal neuroendocrine tumor, squamous cell carcinoma of the skin, astrocytoma, low-grade astrocytoma, high-grade astrocytoma, anaplastic astrocytoma, glioblastoma multiforme.

[0277] In some embodiments, the cancer to be treated / prevented comprises cells expressing EGFR family members (such as HER3, EGFR, HER2, or HER4) and / or cells expressing EGFR family member ligands. In some embodiments, the cancer to be treated / prevented is a cancer positive for EGFR family members. In some embodiments, the cancer comprises cells overexpressing EGFR family members and / or EGFR family member ligands. Overexpression can be identified by detecting expression levels higher than those in equivalent non-cancerous / non-tumor tissue.

[0278] Expression levels can be determined by any suitable method. Expression can be gene expression or protein expression. Gene expression can be determined by detecting the mRNA encoding HER3, for example using quantitative real-time PCR (qRT-PCR). Protein expression can be determined by antibody-based methods, such as Western blotting, immunohistochemistry, immunocytochemistry, flow cytometry, or ELISA.

[0279] In some embodiments, the cancer is HER3 pathology-associated cancer. That is, in some embodiments, the cancer is a cancer caused or aggravated by HER3 expression, a cancer where HER3 expression is a risk factor, and / or a cancer where HER3 expression is positively correlated with the occurrence, development, progression, severity, or metastasis of the cancer. The cancer is characterized by HER3 expression, for example, the cancer contains cells expressing HER3 (i.e., cells of tumor tissue). Such cancers may be referred to as HER3-positive. HER3-positive cancer is cancer that contains cells expressing HER3 (e.g., on the cell surface). HER3-positive cancers may overexpress HER3.

[0280] In some implementations, the cancer to be treated / prevented comprises cells carrying genetic variations (such as mutations) that can lead to increased expression and / or activity of HER3 (gene and / or protein) compared to equivalent cells carrying a reference allele that does not contain genetic variations (such as an unmutated or “wild-type” allele).

[0281] In some implementations, mutations that lead to increased HER3 expression can increase the level of HER3 on or in the cell surface of cells containing the mutation relative to equivalent cells that do not contain the mutation.

[0282] Cells with increased HER3 expression relative to reference cells (e.g., due to mutation) may be termed "overexpressing" HER3 or "upregulated" HER3. For example, a cancer comprising cells carrying a mutation that results in increased HER3 expression relative to equivalent cells lacking this mutation may be termed a cancer comprising cells with HER3 overexpression / upregulated expression. In some embodiments, the reference cells lacking the mutation may be non-cancerous cells (e.g., equivalent cell types) or cancerous cells (e.g., equivalent tumor cells).

[0283] In some embodiments, the cancer to be treated / prevented, as described in this disclosure, is characterized by increased HER3 expression and / or activity (i.e., gene and / or protein expression) in organs / tissues / subjects affected by the disease / symptom compared to normal organs / tissues / subjects (i.e., no disease / symptom). In some embodiments, the cells and / or tumors of the cancer to be treated / prevented are characterized by increased HER3 expression and / or activity compared to the expression and / or activity levels of equivalent non-cancerous / non-tumor tissues.

[0284] As a result of HER3 gene amplification, HER3-overexpressing cancers may overexpress HER3. In some embodiments, the cancer to be treated / prevented, as described in this disclosure, is a HER3-amplified cancer.

[0285] HER3 gene amplification can be identified using techniques well-known in the field, such as in situ hybridization. For example, fluorescence in situ hybridization can be used to assess HER3 amplification; see Chung et al., *Journal of Gynecologic Oncology*, 2019, Vol. 30, No. 5, p. 75.

[0286] In some embodiments, cells containing HER3 amplification contain >2, such as any one or more copies of the HER3 gene from the sequence 3, 4, 5, 6, 7, 8, 9, and 10, or >2, such as any one or more copies of HER3 gene fragments from the sequence 3, 4, 5, 6, 7, 8, 9, and 10. HER3 amplification may be the result of gene duplication or polychromatic amplification. In some embodiments, HER3-amplified cancer may contain a HER3 gene to chromosome 12 ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, HER3-amplified cancer may contain a HER3 gene to chromosome 12 centromere (CEN12) ratio ≥1 (e.g., as determined by ISH), such as ≥1.5, ≥2, or ≥2.5. In some embodiments, HER3-amplified cancer cells may contain HER3 gene copy numbers >2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, or ≥10 (e.g., as determined by NGS).

[0287] For information on HER3 and its relevance and role in cancer, see Mishra et al., *Oncol Rev.*, 2018, Vol. 12, No. 1, p. 355; Karachaliou et al., *BioDrugs*, 2017, Vol. 31, No. 1, pp. 63-73; and Zhang et al., *Acta Biochimica et Biophysica Sinica*, 2016, Vol. 48, No. 1, pp. 39-48. These references are all incorporated herein by reference. Mishra et al., *Oncol Rev.*, 2018, Vol. 12, No. 1, p. 355, also describe targeted interventions for HER3 in cancer treatment, including monoclonal anti-HER3 antibody therapy.

[0288] As described in some aspects and embodiments of this disclosure, the cancer described herein differs from the cancer described in WO2023 / 017151A1. That is, in some embodiments, the cancer described herein is not the cancer described in WO2023 / 017151A1. In some aspects and embodiments, the cancer described herein includes one or more features that distinguish it from the cancer described in WO2023 / 017151A1.

[0289] For illustrative purposes, in some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3) do not contain genetic variations that lead to increased MET expression or enhanced gene product activity (e.g., no MET amplification). Further, in some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3) are cancers containing genetic variations that lead to increased expression or enhanced gene product activity of one or more genes located on chromosomes 3q26-3q28 (such as genes selected from TP63, SOX2, and PIK3CA; or amplification of one or more of TP63, SOX2, and PIK3CA). In some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3) are cancers containing genetic variations that lead to increased expression or enhanced gene product activity of one or more genes located on the short arm of chromosome 7 (such as one or more genes located on chromosome 7p11, such as EGFR; or EGFR amplification). In some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3) are cancers containing genetic variations that result in reduced expression or reduced activity of gene products of one or more genes located on the short arm of chromosome 3 (such as one or more genes located on chromosome 3p21, such as TUSC2, or TUSC2 deletion).

[0290] Conversely, in some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3 in combination with HER3-mediated signal transduction antagonists) contain genetic variations (such as MET amplification) that lead to increased MET expression or enhanced gene product activity. Further, in some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3 in combination with HER3-mediated signal transduction antagonists) do not contain genetic variations that lead to increased expression or enhanced gene product activity of one or more genes located on chromosomes 3q26-3q28 (such as one or more genes selected from TP63, SOX2, and PIK3CA, without amplification of one or more of TP63, SOX2, and PIK3CA). In some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3 in combination with HER3-mediated signal transduction antagonists) do not contain genetic variations that result in increased expression or enhanced gene product activity of one or more genes located on the short arm of chromosome 7 (such as one or more genes located on chromosome 7p11, such as EGFR, if EGFR amplification is not included). In some embodiments, cancers as described in this disclosure (such as cancers treated / prevented using antigen-binding molecules that bind to HER3 in combination with HER3-mediated signal transduction antagonists) do not contain genetic variations that result in decreased expression or reduced gene product activity of one or more genes located on the short arm of chromosome 3 (such as one or more genes located on chromosome 3p21, such as TUSC2, if TUSC2 deletion is not included).

[0291] Cancers characterized by genetic variants that result in enhanced HER3-mediated signaling due to deletion.

[0292] The various aspects and embodiments described in this disclosure relate to cancers lacking genetic variants that lead to enhanced HER3-mediated signaling. Such cancers may be considered more sensitive / more susceptible / less resistant (and therefore more likely to produce a good response) to therapeutic / preventive interventions with antigen-binding molecules that bind to HER3 (e.g., as monotherapy). It should be understood that the cancers described in this section can be further characterized based on the preceding “Cancer” section.

[0293] In the various aspects and embodiments described in this disclosure, cancers to be treated / prevented using antigen-binding molecules that bind to HER3 can be characterized by one or more of the following features:

[0294] (1a)(i) is not a homozygote of the MET activating mutation (the MET activating mutation described in this paper).

[0295] (1a)(ii) are not heterozygous for MET activation mutations (MET activation mutations as described in this article).

[0296] (1a)(iii) Does not include MET amplification (MET amplification as described herein).

[0297] (1a)(iv) do not contain genetic variations that lead to increased MET expression or enhanced activity of gene products.

[0298] (2a)(i) is not a homozygote of the KRAS activating mutation (KRAS activating mutation as described in this article).

[0299] (2a)(ii) are not heterozygous for KRAS activation mutations (KRAS activation mutations as described in this article).

[0300] (2a)(iii) Does not include KRAS amplification (KRAS amplification as described herein).

[0301] (2a)(iv) do not contain genetic variations that lead to increased KRAS expression or enhanced activity of gene products.

[0302] (3a)(i) is not a homozygote of the PIK3CA activating mutation (the PIK3CA activating mutation described in this paper).

[0303] (3a)(ii) are not heterozygous for the PIK3CA activating mutation (the PIK3CA activating mutation described in this paper).

[0304] (3a)(iii) Does not contain PIK3CA amplification (PIK3CA amplification as described herein).

[0305] (3a)(iv) Do not contain genetic variations that lead to increased PIK3CA expression or enhanced activity of gene products.

[0306] (4a)(i) is not a homozygote of the BRAF activating mutation (the BRAF activating mutation described in this article).

[0307] (4a)(ii) are not heterozygous for BRAF activating mutations (BRAF activating mutations as described in this article).

[0308] (4a)(iii) Does not contain BRAF amplification (BRAF amplification as described herein).

[0309] (4a)(iv) do not contain genetic variations that lead to increased BRAF expression or enhanced activity of gene products.

[0310] (5a)(i) does not contain genetic variations that result in reduced PTEN expression or decreased activity of gene products.

[0311] (5a)(ii) are not homozygous for the PTEN inactivation mutation (PTEN inactivation mutation as described in this article).

[0312] (5a)(iii) are not heterozygous for PTEN inactivation mutations (PTEN inactivation mutations as described in this article).

[0313] (5a)(iv) Does not contain PTEN deletion (PTEN deletion as described in this article).

[0314] (6a)(i) includes genetic variations that lead to increased TP63 expression or enhanced activity of the gene product.

[0315] (6a)(ii) includes TP63 amplification (TP63 amplification as described herein).

[0316] (7a)(i) contains genetic variations that lead to increased PIK3CA expression or enhanced activity of the gene product.

[0317] (7a)(ii) includes PIK3CA amplification (PIK3CA amplification as described herein).

[0318] (8a)(i) includes genetic variations that lead to increased SOX2 expression or enhanced activity of gene products.

[0319] (8a)(ii) includes SOX2 amplification (the SOX2 amplification described herein).

[0320] (9a)(i) includes genetic variations that lead to increased expression of the HER3 ligand (as described herein).

[0321] (9a)(ii) includes NRG gene fusions (the NRG gene fusions described herein).

[0322] (9a)(iii) Includes NRG1 gene fusion (the NRG1 gene fusion described herein).

[0323] (10a)(i) includes genetic variations that lead to increased EGFR expression or enhanced activity of gene products.

[0324] (10a)(ii) includes EGFR amplification (EGFR amplification as described herein).

[0325] (11a)(i) does not contain genetic variations that result in reduced TUSC2 expression or decreased activity of gene products.

[0326] (11a)(ii) does not include TUSC2 deletion (the TUSC2 deletion described herein).

[0327] In some implementations, cancer treated / prevented using antigen-binding molecules that bind to HER3 can be characterized by one of the following combinations of features (refer to the preceding paragraph): (1a), (2a); (1a), (3a); (1a), (4a); (1a), (5a); (1a), (6a); (1a), (7a); (1a), (8a); (1a), (9a); (1a), (10a); (1a), (11a); (1a), (2a), (3a); (1a), (2a), (4a); (1a), (2a), (5a); (1a), (2a), (6a); (1a), (2a), (7a); (1a), (2a), (8a); (1a) ), (2a), (9a); (1a), (2a), (10a); (1a), (2a), (11a); (1a), (3a), (4a); (1 a), (3a), (5a); (1a), (3a), (6a); (1a), (3a), (7a); (1a), (3a), (8a); (1a) , (3a), (9a); (1a), (3a), (10a); (1a), (3a), (11a); (1a), (4a), (5a); (1a) , (4a), (6a); (1a), (4a), (7a); (1a), (4a), (8a); (1a), (4a), (9a); (1a), ( 4a), (10a); (1a), (4a), (11a); (1a), (5a), (6a); (1a), (5a), (7a); (1a), (5a), (8a); (1a), (5a), (9a); (1a), (5a), (10a); (1a), (5a), (11a); (1a), (6a), (7a); (1a), (6a), (8a); (1a), (6a), (9a); (1a), (6a), (10a); (1a), ( 6a), (11a); (1a), (7a), (8a); (1a), (7a), (9a); (1a), (7a), (10a); (1a), ( 7a), (11a); (1a), (8a), (9a); (1a), (8a), (10a); (1a), (8a), (11a); (1a) , (9a), (10a); (1a), (9a), (11a); (1a), (10a), (11a); (1a), (2a), (3a), (4 a); (1a), (2a), (3a), (5a); (1a), (2a), (3a), (6a); (1a), (2a), (3a), (7a) ;(1a), (2a), (3a), (8a); (1a), (2a), (3a), (9a); (1a), (2a), (3a), (10a);(1a)、(2a)、(3a)、(11a);(1a)、(2a)、(4a)、(5a);(1a)、(2a)、(4a)、(6a);(1a)、(2a)、(4a)、(7a);(1a)、(2a)、(4a)、(8a);(1a)、(2a)、(4a)、(9a);(1a) (2a), (4a), (10a); (1a), (2a), (4a), (11a); (1a), (2a), (5a), (6a); (1a), (2a), (5a), (7a); (1a), (2a), (5a), (8a); (1a), (2a), (5a), (9a); (1a), (2 a) (5a) (10a) (1a) (2a) (5a) (11a) (1a) (2a) (6a) (7a) (1a) (2a) (6a) (8a) (1a) (2a) (6a) (9a) (1a) (2a) (6a) (10a) (1a) (2a) (6a) (11a); (1a) (2a) (7a) (8a); (1a) (2a) (7a) (9a); (1a) (2a) (7a) (10a); (1a) (2a) (7a) (11a); (1a) (2a) (8a) (9a); (1a) (2a) ( 8a)、(10a);(1a)、(2a)、(8a)、(11a);(1a)、(2a)、(9a)、(10a);(1a)、(2a)、(9a)、(11a);(1a)、(2a)、(10a)、(11a);(1a)、(3a)、(4a)、(5a);(1a)、(3a)、 (4a)、(6a);(1a)、(3a)、(4a)、(7a);(1a)、(3a)、(4a)、(8a);(1a)、(3a)、(4a)、(9a);(1a)、(3a)、(4a)、(10a);(1a)、(3a)、(4a)、(11a);(1a)、(3a)、(5a )、(6a);(1a)、(3a)、(5a)、(7a);(1a)、(3a)、(5a)、(8a);(1a)、(3a)、(5a)、(9a);(1a)、(3a)、(5a)、(10a);(1a)、(3a)、(5a)、(11a);(1a)、(3a)、(6a)、( 7a); (1a), (3a), (6a), (8a); (1a), (3a), (6a), (9a); (1a), (3a), (6a), (10a); (1a), (3a), (6a), (11a); (1a), (3a), (7a), (8a); (1a), (3a), (7a), (9a);(1a)、(3a)、(7a)、(10a);(1a)、(3a)、(7a)、(11a);(1a)、(3a)、(8a)、(9a);(1a)、(3a)、(8a)、(10a);(1a)、(3a)、(8a)、(11a);(1a)、(3a)、(9a)、(10a) (1a), (3a), (9a), (11a); (1a), (3a), (10a), (11a); (1a), (4a), (5a), (6a); (1a), (4a), (5a), (7a); (1a), (4a), (5a), (8a); (1a), (4a), (5a), (9a) (1a)、(4a)、(5a)、(10a);(1a)、(4a)、(5a)、(11a);(1a)、(4a)、(6a)、(7a); (1a)、(4a)、(6a)、(8a);(1a)、(4a)、(6a)、(9a);(1a)、(4a)、(6a)、(10a);( 1a)、(4a)、(6a)、(11a);(1a)、(4a)、(7a)、(8a);(1a)、(4a)、(7a)、(9a);(1 a)、(4a)、(7a)、(10a);(1a)、(4a)、(7a)、(11a);(1a)、(4a)、(8a)、(9a);(1 a)、(4a)、(8a)、(10a);(1a)、(4a)、(8a)、(11a);(1a)、(4a)、(9a)、(10a);( 1a)、(4a)、(9a)、(11a);(1a)、(4a)、(10a)、(11a);(1a)、(5a)、(6a)、(7a); (1a)、(5a)、(6a)、(8a);(1a)、(5a)、(6a)、(9a);(1a)、(5a)、(6a)、(10a);(1a)、(5a)、(6a)、(11a);(1a)、(5a)、(7a)、(8a);(1a)、(5a)、(7a)、(9a);(1 a)、(5a)、(7a)、(10a);(1a)、(5a)、(7a)、(11a);(1a)、(5a)、(8a)、(9a);(1a)、(5a)、(8a)、(10a);(1a)、(5a)、(8a)、(11a);(1a)、(5a)、(9a)、(10a);( 1a)、(5a)、(9a)、(11a);(1a)、(5a)、(10a)、(11a);(1a)、(6a)、(7a)、(8a);(1a)、(6a)、(7a)、(9a);(1a)、(6a)、(7a)、(10a);(1a)、(6a)、(7a)、(11a);(1a)、(6a)、(8a)、(9a);(1a)、(6a)、(8a)、(10a);(1a)、(6a)、(8a)、(11a); (1a)、(6a)、(9a)、(10a);(1a)、(6a)、(9a)、(11a);(1a)、(6a)、(10a)、(11a) (1a), (7a), (8a), (9a); (1a), (7a), (8a), (10a); (1a), (7a), (8a), (11a); (1a), (7a), (9a), (10a); (1a), (7a), (9a), (11a); (1a), (7a), (10a), (11a) a); (1a), (8a), (9a), (10a); (1a), (8a), (9a), (11a); (1a), (8a), (10a), (11a); (1a), (9a), (10a), (11a); (1a), (2a), (3a), (4a), (5a); (1a), (2a) 3a), (4a), (6a); (1a), (2a), (3a), (4a), (7a); (1a), (2a), (3a), (4a), (8a); (1a), (2a), (3a), (4a), (9a); (1a), (2a), (3a), (4a), (10a); (1a), (2a) (3a)、(4a)、(11a);(1a)、(2a)、(3a)、(5a)、(6a);(1a)、(2a)、(3a)、(5a)、(7a);(1a)、(2a)、(3a)、(5a)、(8a);(1a)、(2a)、(3a)、(5a)、(9a);(1a)、(2a) (3a), (5a), (10a); (1a), (2a), (3a), (5a), (11a); (1a), (2a), (3a), (6a), (7a); (1a), (2a), (3a), (6a), (8a); (1a), (2a), (3a), (6a), (9a); (1a), (2 a) (3a) (6a) (10a) (1a) (2a) (3a) (6a) (11a) (1a) (2a) (3a) (7a) (8a) (1a) (2a) (3a) (7a) (9a) (1a) (2a) (3a) (7a) (10a) (1a) (2a), (3a), (7a), (11a); (1a), (2a), (3a), (8a), (9a); (1a), (2a), (3a), (8a), (10a); (1a), (2a), (3a), (8a), (11a); (1a), (2a), (3a), (9a), (10a);(1a), (2a), (3a), (9a), (11a); (1a), (2a), (3a), (10a), (11a); (1a), (2a), (4a), (5a), (6a); (1a), (2a), (4a), (5a), (7a); (1a), (2a), (4a), (5a), (8 a) (1a), (2a), (4a), (5a), (9a) (1a), (2a), (4a), (5a), (10a) (1a), (2a), (4a), (5a), (11a) (1a), (2a), (4a), (6a), (7a) (1a), (2a), (4a), (6a) (8a); (1a), (2a), (4a), (6a), (9a); (1a), (2a), (4a), (6a), (10a); (1a), (2a), (4a), (6a), (11a); (1a), (2a), (4a), (7a), (8a); (1a), (2a), (4a), (7a (9a); (1a); (2a); (4a); (7a); (10a); (1a); (2a); (4a); (7a); (11a); (1a); (2a); (4a); (8a); (9a); (1a); (2a); (4a); (8a); (10a); (1a); (2a); (4a) (8a)、(11a);(1a)、(2a)、(4a)、(9a)、(10a);(1a)、(2a)、(4a)、(9a)、(11a);(1a)、(2a)、(4a)、(10a)、(11a);(1a)、(2a)、(5a)、(6a)、(7a);(1a)、(2a)、 (5a), (6a), (8a); (1a), (2a), (5a), (6a), (9a); (1a), (2a), (5a), (6a), (10a); (1a), (2a), (5a), (6a), (11a); (1a), (2a), (5a), (7a), (8a); (1a), (2a (5a), (7a), (9a); (1a), (2a), (5a), (7a), (10a); (1a), (2a), (5a), (7a), (11a); (1a), (2a), (5a), (8a), (9a); (1a), (2a), (5a), (8a), (10a); (1a) (2a), (5a), (8a), (11a); (1a), (2a), (5a), (9a), (10a); (1a), (2a), (5a), (9a), (11a); (1a), (2a), (5a), (10a), (11a); (1a), (2a), (6a), (7a), (8a);(1a)、(2a)、(6a)、(7a)、(9a);(1a)、(2a)、(6a)、(7a)、(10a);(1a)、(2a)、(6a)、(7a)、(11a);(1a)、(2a)、(6a)、(8a)、(9a);(1a)、(2a)、(6a)、(8a)、 (10a); (1a), (2a), (6a), (8a), (11a); (1a), (2a), (6a), (9a), (10a); (1a), (2a), (6a), (9a), (11a); (1a), (2a), (6a), (10a), (11a); (1a), (2a), ( (7a), (8a), (9a); (1a), (2a), (7a), (8a), (10a); (1a), (2a), (7a), (8a), (11a); (1a), (2a), (7a), (9a), (10a); (1a), (2a), (7a), (9a), (11a); (1a) (2a), (7a), (10a), (11a); (1a), (2a), (8a), (9a), (10a); (1a), (2a), (8a), (9a), (11a); (1a), (2a), (8a), (10a), (11a); (1a), (2a), (9a), (10a) (11a); (1a), (3a), (4a), (5a), (6a); (1a), (3a), (4a), (5a), (7a); (1a), (3a), (4a), (5a), (8a); (1a), (3a), (4a), (5a), (9a); (1a), (3a), (4a) (5a)、(10a);(1a)、(3a)、(4a)、(5a)、(11a);(1a)、(3a)、(4a)、(6a)、(7a);(1a)、(3a)、(4a)、(6a)、(8a);(1a)、(3a)、(4a)、(6a)、(9a);(1a)、(3a)、(4 a)、(6a)、(10a);(1a)、(3a)、(4a)、(6a)、(11a);(1a)、(3a)、(4a)、(7a)、(8a);(1a)、(3a)、(4a)、(7a)、(9a);(1a)、(3a)、(4a)、(7a)、(10a);(1a)、( 3a)、(4a)、(7a)、(11a);(1a)、(3a)、(4a)、(8a)、(9a);(1a)、(3a)、(4a)、( 8a)、(10a);(1a)、(3a)、(4a)、(8a)、(11a);(1a)、(3a)、(4a)、(9a)、(10a);(1a)、(3a)、(4a)、(9a)、(11a);(1a)、(3a)、(4a)、(10a)、(11a);(1a)、(3a)、(5a)、(6a)、(7a);(1a)、(3a)、(5a)、(6a)、(8a);(1a)、(3a)、(5a)、(6a) (9a); (1a); (3a); (5a); (6a); (10a); (1a); (3a); (5a); (6a); (11a); (1a); (3a); (5a); (7a); (8a); (1a); (3a); (5a); (7a); (9a); (1a); (3a); (5a) (7a)、(10a);(1a)、(3a)、(5a)、(7a)、(11a);(1a)、(3a)、(5a)、(8a)、(9a);(1a)、(3a)、(5a)、(8a)、(10a);(1a)、(3a)、(5a)、(8a)、(11a);(1a)、(3a) (5a), (9a), (10a); (1a), (3a), (5a), (9a), (11a); (1a), (3a), (5a), (10a), (11a); (1a), (3a), (6a), (7a), (8a); (1a), (3a), (6a), (7a), (9a); (1a (3a), (6a), (7a), (10a); (1a), (3a), (6a), (7a), (11a); (1a), (3a), (6a), (8a), (9a); (1a), (3a), (6a), (8a), (10a); (1a), (3a), (6a), (8a), (11a) a); (1a), (3a), (6a), (9a), (10a); (1a), (3a), (6a), (9a), (11a); (1a), (3a), (6a), (10a), (11a); (1a), (3a), (7a), (8a), (9a); (1a), (3a), (7a) (8a)、(10a);(1a)、(3a)、(7a)、(8a)、(11a);(1a)、(3a)、(7a)、(9a)、(10a);(1a)、(3a)、(7a)、(9a)、(11a);(1a)、(3a)、(7a)、(10a)、(11a);(1a)、(3a (8a), (9a), (10a); (1a), (3a), (8a), (9a), (11a); (1a), (3a), (8a), (10a), (11a); (1a), (3a), (9a), (10a), (11a); (1a), (4a), (5a), (6a), (7a);(1a)、(4a)、(5a)、(6a)、(8a);(1a)、(4a)、(5a)、(6a)、(9a);(1a)、(4a)、( 5a)、(6a)、(10a);(1a)、(4a)、(5a)、(6a)、(11a);(1a)、(4a)、(5a)、(7a)、 (8a); (1a), (4a), (5a), (7a), (9a); (1a), (4a), (5a), (7a), (10a); (1a), (4a), (5a), (7a), (11a); (1a), (4a), (5a), (8a), (9a); (1a), (4a), (5a) 8a)、(10a);(1a)、(4a)、(5a)、(8a)、(11a);(1a)、(4a)、(5a)、(9a)、(10a) ;(1a)、(4a)、(5a)、(9a)、(11a);(1a)、(4a)、(5a)、(10a)、(11a);(1a)、(4a )、(6a)、(7a)、(8a);(1a)、(4a)、(6a)、(7a)、(9a);(1a)、(4a)、(6a)、(7a) 、(10a);(1a)、(4a)、(6a)、(7a)、(11a);(1a)、(4a)、(6a)、(8a)、(9a);(1a) 、(4a)、(6a)、(8a)、(10a);(1a)、(4a)、(6a)、(8a)、(11a);(1a)、(4a)、(6a) )、(9a)、(10a);(1a)、(4a)、(6a)、(9a)、(11a);(1a)、(4a)、(6a)、(10a)、( 11a);(1a)、(4a)、(7a)、(8a)、(9a);(1a)、(4a)、(7a)、(8a)、(10a);(1a)、 (4a)、(7a)、(8a)、(11a);(1a)、(4a)、(7a)、(9a)、(10a);(1a)、(4a)、(7a)、 (9a)、(11a);(1a)、(4a)、(7a)、(10a)、(11a);(1a)、(4a)、(8a)、(9a)、(10 a);(1a)、(4a)、(8a)、(9a)、(11a);(1a)、(4a)、(8a)、(10a)、(11a);(1a)、( 4a), (9a), (10a), (11a); (1a), (5a), (6a), (7a), (8a); (1a), (5a), (6a), (7a), (9a); (1a), (5a), (6a), (7a), (10a); (1a), (5a), (6a), (7a), (11a);(1a)、(5a)、(6a)、(8a)、(9a);(1a)、(5a)、(6a)、(8a)、(10a);(1a)、(5a)、(6a)、(8a)、(11a);(1a)、(5a)、(6a)、(9a)、(10a);(1a)、(5a)、(6a)、(9a)、( (11a); (1a), (5a), (6a), (10a), (11a); (1a), (5a), (7a), (8a), (9a); (1a), (5a), (7a), (8a), (10a); (1a), (5a), (7a), (8a), (11a); (1a), (5a), (7a) (9a)、(10a);(1a)、(5a)、(7a)、(9a)、(11a);(1a)、(5a)、(7a)、(10a)、(11a);(1a)、(5a)、(8a)、(9a)、(10a);(1a)、(5a)、(8a)、(9a)、(11a);(1a)、(5a (8a), (10a), (11a); (1a), (5a), (9a), (10a), (11a); (1a), (6a), (7a), (8a), (9a); (1a), (6a), (7a), (8a), (10a); (1a), (6a), (7a), (8a), (11a); (1 a)、(6a)、(7a)、(9a)、(10a);(1a)、(6a)、(7a)、(9a)、(11a);(1a)、(6a)、(7a)、(10a)、(11a);(1a)、(6a)、(8a)、(9a)、(10a);(1a)、(6a)、(8a)、(9a)、( (11a); (1a), (6a), (8a), (10a), (11a); (1a), (6a), (9a), (10a), (11a); (1a), (7a), (8a), (9a), (10a); (1a), (7a), (8a), (9a), (11a); (1a), (7a), (8a) (10a), (11a); (1a), (7a), (9a), (10a), (11a); (1a), (8a), (9a), (10a), (11a); (1a), (2a), (3a), (4a), (5a), (6a); (1a), (2a), (3a), (4a), (5a), (7a) a); (1a), (2a), (3a), (4a), (5a), (8a); (1a), (2a), (3a), (4a), (5a), (9a); (1a), (2a), (3a), (4a), (5a), (10a); (1a), (2a), (3a), (4a), (5a), (11a);(1a)、(2a)、(3a)、(4a)、(6a)、(7a);(1a)、(2a)、(3a)、(4a)、(6a)、(8a);(1a)、(2a)、(3a)、(4a)、(6a)、(9a);(1a)、(2a)、(3a)、(4a)、(6a)、(10a);( (1a) (2a) (3a) (4a) (6a) (11a) (1a) (2a) (3a) (4a) (7a) (8a) (1a) (2a) (3a) (4a) (7a) (9a) (1a) (2a) (3a) (4a) (7a) (10a) (1 a)、(2a)、(3a)、(4a)、(7a)、(11a);(1a)、(2a)、(3a)、(4a)、(8a)、(9a);(1a)、(2a)、(3a)、(4a)、(8a)、(10a);(1a)、(2a)、(3a)、(4a)、(8a)、(11a);( (1a), (2a), (3a), (4a), (9a), (10a); 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(1a), (2a), (4a), (8a), (9a), (10a); (1a), (2a), (4a), (8a), (9a), (11a); (1a), (2a), (4a), (8a), (10a), (11a) (1a), (2a), (4a), (9a), (10a), (11a); (1a), (2a), (5a), (6a), (7a), (8a); (1a), (2a), (5a), (6a), (7a), (9a); (1a), (2a), (5a), (6a), (7a), (10a) (1a), (2a), (5a), (6a), (7a), (11a); (1a), (2a), (5a), (6a), (8a), (9a); (1a), (2a), (5a), (6a), (8a), (10a); (1a), (2a), (5a), (6a), (8a), (11a) (1a), (2a), (5a), (6a), (9a), (10a); (1a), (2a), (5a), (6a), (9a), (11a); (1a), (2a), (5a), (6a), (10a), (11a); (1a), (2a), (5a), (7a), (8a), (9a);(1a), (2a), (5a), (7a), (8a), (10a); (1a), (2a), (5a), (7a), (8a), (11a); (1a), (2a), (5a), (7a), (9a), (10a); (1a), (2a), (5a), (7a), (9a), (11a); (1a), (2a), (5a), (7a), (10a), (11a); (1a), (2a), (5a), (8a), (9a), (10a); (1a), (2a), (5a), (8a), (9a), (11a); (1a), (2a), (5a), (8a), (10a), (11a (1a), (2a), (5a), (9a), (10a), (11a); (1a), (2a), (6a), (7a), (8a), (9a); (1a), (2a), (6a), (7a), (8a), (10a); (1a), (2a), (6a), (7a), (8a), (11a) (1a), (2a), (6a), (7a), (9a), (10a); 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(1a), (2a), (3a), (5a), (6a), (7a), (8a), (9a), (11a); (1a) (2a), (3a), (5a), (6a), (7a), (8a), (10a), (11a); (1a), (2a), (3a), (5a), (6a), (7a), (9a), (10a), (11a); (1a), (2a), (3a), (5a), (6a), (8a), (9a) (10a)、(11a);(1a)、(2a)、(3a)、(5a)、(7a)、(8a)、(9a)、(10a)、(11a);(1a)、(2a)、(3a)、(6a)、(7a)、(8a)、(9a)、(10a)、(11a);(1a)、(2a)、(4a)、( (5a), (6a), (7a), (8a), (9a), (10a); (1a), (2a), (4a), (5a), (6a), (7a), (8a), (9a), (11a); (1a), (2a), (4a), (5a), (6a), (7a), (8a), (10a), (11a);(1a)、(2a)、(4a)、(5a)、(6a)、(7a)、(9a)、(10a)、(11a);(1a)、(2a)、(4a)、(5a)、(6a)、(8a)、(9a)、(10a)、(11a);(1a)、(2a)、(4a)、(5a)、(7a)、( (8a), (9a), (10a), (11a); (1a), (2a), (4a), (6a), (7a), (8a), (9a), (10a), (11a); (1a), (2a), (5a), (6a), (7a), (8a), (9a), (10a), (11a); (1a) (3a), (4a), (5a), (6a), (7a), (8a), (9a), (10a); (1a), (3a), (4a), (5a), (6a), (7a), (8a), (9a), (11a); (1a), (3a), (4a), (5a), (6a), (7a), (8a) (10a)、(11a);(1a)、(3a)、(4a)、(5a)、(6a)、(7a)、(9a)、(10a)、(11a);(1a)、(3a)、(4a)、(5a)、(6a)、(8a)、(9a)、(10a)、(11a);(1a)、(3a)、(4a)、 (5a), (7a), (8a), (9a), (10a), (11a); (1a), (3a), (4a), (6a), (7a), (8a), (9a), (10a), (11a); (1a), (3a), (5a), (6a), (7a), (8a), (9a), (10a) (11a); (1a), (4a), (5a), (6a), (7a), (8a), (9a), (10a), (11a); (1a), (2a), (3a), (4a), (5a), (6a), (7a), (8a), (9a), (10a); (1a), (2a), (3a), (4a) a) (5a) (6a) (7a) (8a) (9a) (11a) (1a) (2a) (3a) (4a) (5a) (6a) (7a) (8a) (10a) (11a) (1a) (2a) (3a) (4a) (5a) (6a) (7a) (9a), (10a), (11a); (1a), (2a), (3a), (4a), (5a), (6a), (8a), (9a), (10a), (11a); (1a), (2a), (3a), (4a), (5a), (7a), (8a), (9a), (10a), (11a);(1a), (2a), (3a), (4a), (6a), (7a), (8a), (9a), (10a), (11a); (1a), (2a), (3a) , (5a), (6a), (7a), (8a), (9a), (10a), (11a); (1a), (2a), (4a), (5a), (6a), (7a ), (8a), (9a), (10a), (11a); (1a), (3a), (4a), (5a), (6a), (7a), (8a), (9a), (1 0a), (11a); (1a), (2a), (3a), (4a), (5a), (6a), (7a), (8a), (9a), (10a), (11a). ;

[0328] In the combinations described above, “(1a)” is independently selected from (1a)(i), (1a)(ii), (1a)(iii), and (1a)(iv) above; and “(2a)” is independently selected from (2a)(i), (2a)(ii), (2a)(iii), and (2a)(iv) above; and “(3a)” is independently selected from (3a)(i), (3a)(ii), (3a)(iii), and (3a)(iv) above; and “(4a)” is independently selected from (4a)(i), (4a)(ii), and (4a)(i) above. (ii) and (4a)(iv); and “(5a)” is independently selected from (5a)(i), (5a)(ii), (5a)(iii) and (5a)(iv) above; and “(6a)” is independently selected from (6a)(i) and (6a)(ii) above; and “(7a)” is independently selected from (7a)(i) and (7a)(ii) above; and “(8a)” is independently selected from (8a)(i) and (8a)(ii) above; and “(9a)” is independently selected from (9a)(i), (9a)(ii) and (9a)(iii) above.

[0329] All possible combinations of the features described above are included. For illustrative purposes only, cancers containing the feature combinations “(1a), (2a)” specifically cover cancers containing the following features: (1a)(i) and (2a)(i); (1a)(i) and (2a)(ii); (1a)(i) and (2a)(iii); (1a)(i) and (2a)(iv); (1a)(ii) and (2a)(i); (1a)(ii) and (2a)(ii); (1a)(ii) and (2a)(iii); (1a)(ii) and (2a)(iii); (1a)(ii) and (2a)(iii); (1a)(ii) and (2a)(iii); (1a)(ii) and (2a)(iii); (1a)(ii) and (2a)(iii); (1a)(ii) and (2a)(iii) a)(ii) and (2a)(iv); (1a)(iii) and (2a)(i); (1a)(iii) and (2a)(ii); (1a)(iii) and (2a)(iii); (1a)(iii) and (2a)(iv); (1a)(iv) and (2a)(i); (1a)(iv) and (2a)(ii); (1a)(iv) and (2a)(iii); and (1a)(iv) and (2a)(iv).

[0330] In embodiments of cancer characterized according to (7a)(i) or (7a)(ii) above, it can no longer be characterized according to (3a)(iii) or (3a)(iv) above. Conversely, in embodiments of cancer characterized according to (3a)(iii) or (3a)(iv) above, it can no longer be characterized according to (7a)(i) or (7a)(ii) above. However, in some embodiments, cancer characterized by (3a)(i) or (3a)(ii) above can be further characterized by (7a)(i) or (7a)(ii) above.

[0331] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by the HER3-binding antigen-binding molecule is not homozygous for activating mutations of the following genes encoding positive regulators of HER3-mediated signal transduction: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer is not a heterozygote selected from the following genes encoding positive regulators of HER3-mediated signal transduction: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer does not contain genetic variations that result in increased expression of genes or enhanced activity of gene products encoding HER3-mediated positive regulators of signal transduction selected from the following: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5.

[0332] In this document, it should be understood that the term “gene” refers to “at least one gene” and includes “one or more genes”. Therefore, a “gene” selected from the given list can be (depending on the number of genes listed in the table) one gene from the table, or any one or more of 2, 3, 4, 5, 6, 7, 8, 9, 10 genes, or all genes listed in the list.

[0333] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by the HER3-binding antigen-binding molecule is not homozygous for activating mutations in genes encoding positive regulators of the MAPK / ERK pathway signaling: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1. In some embodiments, the cancer is not a heterozygote for activating mutations in genes selected from the following gene encoding positive regulators of MAPK / ERK pathway signaling: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1. In some embodiments, the cancer does not contain genetic variations selected from the following that result in increased expression of genes encoding positive regulators of the MAPK / ERK pathway signaling pathway or enhanced activity of gene products: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1.

[0334] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by the HER3-binding antigen-binding molecule is not homozygous for activating mutations in genes encoding positive regulators of the PI3K / AKT / mTOR pathway signaling: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer is not a heterozygote of an activating mutation of a gene encoding a positive regulator of the PI3K / AKT / mTOR pathway: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer does not contain genetic variations selected from the following that lead to increased expression of genes encoding positive regulators of the PI3K / AKT / mTOR pathway or enhanced activity of gene products: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5.

[0335] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule is not homozygous for an inactivating mutation of a gene encoding a negative regulator of HER3-mediated signal transduction, selected from the following: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the cancer is not heterozygous for an inactivating mutation of a gene encoding a negative regulator of HER3-mediated signal transduction, selected from the following: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the cancer does not contain a genetic variant selected from the following that results in reduced expression of a gene encoding a negative regulator of HER3-mediated signal transduction or reduced activity of its gene product: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0336] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using antigen-binding molecules binding to HER3 is not homozygous for an NF1 gene inactivation mutation. In some embodiments, the cancer of the non-inactivation mutation heterozygote is not heterozygous for an NF1 gene inactivation mutation. In some embodiments, the cancer that does not contain genetic variations that result in reduced expression or decreased activity of the gene product is not homozygous for an NF1 inactivation mutation.

[0337] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule is not homozygous for gene inactivation mutations encoding negative regulators of the PI3K / AKT / mTOR pathway, selected from the following: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1. In some embodiments, the cancer is not heterozygous for gene inactivation mutations encoding negative regulators of the PI3K / AKT / mTOR pathway, selected from the following: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1. In some embodiments, the cancer does not contain genetic variations selected from the following that result in reduced expression or decreased activity of gene products encoding negative regulators of the PI3K / AKT / mTOR pathway: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1.

[0338] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity. In some embodiments, the cancer (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations.

[0339] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain PIK3CA activating mutations.

[0340] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer (i) does not contain MET amplification, and (ii) does not contain BRAF activating mutations.

[0341] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that result in increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that result in decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain PTEN inactivation mutations.

[0342] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain PIK3CA activating mutations.

[0343] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain BRAF activating mutations.

[0344] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain PTEN inactivating mutations.

[0345] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain PIK3CA activating mutations, and (iii) does not contain BRAF activating mutations.

[0346] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain PIK3CA activating mutations, and (iii) does not contain PTEN inactivating mutations.

[0347] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain BRAF activating mutations, and (iii) does not contain PTEN inactivating mutations.

[0348] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iv) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain PIK3CA activating mutations, and (iv) does not contain BRAF activating mutations.

[0349] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iv) does not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain PIK3CA activating mutations, and (iv) does not contain PTEN inactivating mutations.

[0350] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity, and (iv) does not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain BRAF activating mutations, and (iv) does not contain PTEN inactivating mutations.

[0351] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity, and (iv) does not contain genetic variants that lead to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain PIK3CA activating mutations, and (iii) does not contain BRAF activating mutations, and (iv) does not contain PTEN inactivating mutations.

[0352] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using the HER3-binding antigen-binding molecule: (i) does not contain genetic variants that lead to increased MET expression or enhanced gene product activity, and (ii) does not contain genetic variants that lead to increased KRAS expression or enhanced gene product activity, and (iii) does not contain genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iv) does not contain genetic variants that lead to increased BRAF expression or enhanced gene product activity, and (v) does not contain genetic variants that lead to decreased PTEN expression or decreased gene product activity. In some embodiments, the cancer: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain PIK3CA activating mutations, and (iv) does not contain BRAF activating mutations, and (v) does not contain PTEN inactivating mutations.

[0353] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by the antigen-binding molecule that binds to HER3: (i) does not contain MET amplification, and (ii) does not contain KRAS activating mutations, and (iii) does not contain PIK3CA activating mutations, and (iv) does not contain BRAF activating mutations, and (v) does not contain PTEN deletions.

[0354] Cancers characterized by the presence of genetic variants that lead to enhanced HER3-mediated signaling.

[0355] The various aspects and embodiments described in this disclosure relate to cancers containing genetic variations that lead to enhanced HER3-mediated signaling. Such cancers are considered to be less sensitive / less susceptible / more resistant to therapeutic / preventive interventions using HER3-binding antigen-binding molecules as monotherapy (and therefore may respond poorly). Administration of HER3-mediated signaling antagonists may effectively make the cancer sensitive / susceptible to therapeutic / preventive interventions using HER3-binding antigen-binding molecules (and thus respond better). It should be understood that the cancers described in this section may be further characterized according to the features described in the preceding section on “Cancer”.

[0356] In the various aspects and embodiments described in this disclosure, cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist can be characterized by one or more of the following features:

[0357] (1b)(i) is a homozygote of the MET activation mutation (the MET activation mutation described in this paper).

[0358] (1b)(ii) are heterozygotes for the MET activation mutation (the MET activation mutation described in this paper).

[0359] (1b)(iii) Includes MET amplification (MET amplification as described herein).

[0360] (1b)(iv) Includes genetic variations that lead to increased MET expression or enhanced activity of gene products.

[0361] (2b)(i) is a homozygote of the KRAS activation mutation (the KRAS activation mutation described in this paper).

[0362] (2b)(ii) are heterozygotes for KRAS activation mutations (KRAS activation mutations as described in this paper).

[0363] (2b)(iii) Includes KRAS amplification (KRAS amplification as described herein).

[0364] (2b)(iv) Includes genetic variations that lead to increased KRAS expression or enhanced activity of gene products.

[0365] (3b)(i) is a homozygote of the PIK3CA activating mutation (the PIK3CA activating mutation described in this paper).

[0366] (3b)(ii) are heterozygotes of the PIK3CA activating mutation (the PIK3CA activating mutation described in this paper).

[0367] (3b)(iii) Includes PIK3CA amplification (PIK3CA amplification as described herein).

[0368] (3b)(iv) Includes genetic variations that lead to increased PIK3CA expression or enhanced activity of the gene product.

[0369] (4b)(i) is a homozygote of the BRAF activating mutation (the BRAF activating mutation described in this paper).

[0370] (4b)(ii) is a heterozygote of the BRAF activation mutation (the BRAF activation mutation described in this paper).

[0371] (4b)(iii) Includes BRAF amplification (BRAF amplification as described herein).

[0372] (4b)(iv) Includes genetic variations that lead to increased BRAF expression or enhanced activity of gene products.

[0373] (5b)(i) includes genetic variations that result in reduced PTEN expression or decreased activity of gene products.

[0374] (5b)(ii) is a homozygote of the PTEN inactivation mutation (PTEN inactivation mutation as described in this article).

[0375] (5b)(iii) is a heterozygote of the PTEN inactivation mutation (PTEN inactivation mutation as described in this article).

[0376] (5b)(iv) Contains PTEN deletion (PTEN deletion as described in this article).

[0377] (6b)(i) includes genetic variations that lead to increased TP63 expression or enhanced activity of the gene product.

[0378] (6b)(ii) includes TP63 amplification (TP63 amplification as described herein).

[0379] (7b)(i) includes genetic variations that lead to increased SOX2 expression or enhanced activity of gene products.

[0380] (7b)(ii) includes SOX2 amplification (the SOX2 amplification described herein).

[0381] (8b)(i) includes genetic variations that lead to increased expression of the HER3 ligand (as described herein).

[0382] (8b)(ii) includes NRG gene fusions (the NRG gene fusions described herein).

[0383] (8b)(iii) Includes NRG1 gene fusion (the NRG1 gene fusion described herein).

[0384] (9b)(i) includes genetic variations that lead to increased EGFR expression or enhanced activity of gene products.

[0385] (9b)(ii) includes EGFR amplification (EGFR amplification as described herein).

[0386] (10b)(i) does not contain genetic variations that result in reduced TUSC2 expression or decreased activity of the gene product.

[0387] (10b)(ii) does not include TUSC2 deletion (the TUSC2 deletion described herein).

[0388] In some implementations, cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist can be characterized by one of the following combinations of features (see previous paragraph): (1b), (2b); (1b), (3b); (1b), (4b); (1b), (5b); (1b), (6b); (1b), (7b); (1b), (8b); (1b), (9b); (1b), (10b); (1b), (2b), (3b); (1b), (2b), (4b); (1b), (2b), (5b); (1b), (2b), (6b); (1b), (2b), (7b); (1b), (2b), ( 8b); (1b), (2b), (9b); (1b), (2b), (10b); (1b), (3b), (4b); (1b), (3b), (5 b); (1b), (3b), (6b); (1b), (3b), (7b); (1b), (3b), (8b); (1b), (3b), (9b) ;(1b), (3b), (10b); (1b), (4b), (5b); (1b), (4b), (6b); (1b), (4b), (7b); (1b), (4b), (8b); (1b), (4b), (9b); (1b), (4b), (10b); (1b), (5b), (6b); ( 1b), (5b), (7b); (1b), (5b), (8b); (1b), (5b), (9b); (1b), (5b), (10b); (1 b), (6b), (7b); (1b), (6b), (8b); (1b), (6b), (9b); (1b), (6b), (10b); (1b) ), (7b), (8b); (1b), (7b), (9b); (1b), (7b), (10b); (1b), (8b), (9b); (1b) , (8b), (10b); (1b), (9b), (10b); (1b), (2b), (3b), (4b); (1b), (2b), (3b) , (5b); (1b), (2b), (3b), (6b); (1b), (2b), (3b), (7b); (1b), (2b), (3b), ( 8b); (1b), (2b), (3b), (9b); (1b), (2b), (3b), (10b); (1b), (2b), (4b), (5 b); (1b), (2b), (4b), (6b); (1b), (2b), (4b), (7b); (1b), (2b), (4b), (8b) ;(1b), (2b), (4b), (9b); (1b), (2b), (4b), (10b); (1b), (2b), (5b), (6b);(1b)、(2b)、(5b)、(7b);(1b)、(2b)、(5b)、(8b);(1b)、(2b)、(5b)、(9b);(1 b)、(2b)、(5b)、(10b);(1b)、(2b)、(6b)、(7b);(1b)、(2b)、(6b)、(8b);(1b )、(2b)、(6b)、(9b);(1b)、(2b)、(6b)、(10b);(1b)、(2b)、(7b)、(8b);(1b) 、(2b)、(7b)、(9b);(1b)、(2b)、(7b)、(10b);(1b)、(2b)、(8b)、(9b);(1b)、 (2b)、(8b)、(10b);(1b)、(2b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b);(1b)、(3b)、(4b)、(6b);(1b)、(3b)、(4b)、(7b);(1b)、(3b)、(4b)、(8b);(1b)、(3 b)、(4b)、(9b);(1b)、(3b)、(4b)、(10b);(1b)、(3b)、(5b)、(6b);(1b)、(3b)、(5b)、(7b);(1b)、(3b)、(5b)、(8b);(1b)、(3b)、(5b)、(9b);(1b)、(3b)、( 5b)、(10b);(1b)、(3b)、(6b)、(7b);(1b)、(3b)、(6b)、(8b);(1b)、(3b)、(6b)、(9b);(1b)、(3b)、(6b)、(10b);(1b)、(3b)、(7b)、(8b);(1b)、(3b)、(7b) )、(9b);(1b)、(3b)、(7b)、(10b);(1b)、(3b)、(8b)、(9b);(1b)、(3b)、(8b)、(10b);(1b)、(3b)、(9b)、(10b);(1b)、(4b)、(5b)、(6b);(1b)、(4b)、(5b)、 (7b);(1b)、(4b)、(5b)、(8b);(1b)、(4b)、(5b)、(9b);(1b)、(4b)、(5b)、(10b);(1b)、(4b)、(6b)、(7b);(1b)、(4b)、(6b)、(8b);(1b)、(4b)、(6b)、(9b );(1b)、(4b)、(6b)、(10b);(1b)、(4b)、(7b)、(8b);(1b)、(4b)、(7b)、(9b);(1b)、(4b)、(7b)、(10b);(1b)、(4b)、(8b)、(9b);(1b)、(4b)、(8b)、(10b);(1b)、(4b)、(9b)、(10b);(1b)、(5b)、(6b)、(7b);(1b)、(5b)、(6b)、(8b);(1b)、(5b)、(6b)、(9b);(1b)、(5b)、(6b)、(10b);(1b)、(5b)、(7b)、(8b);( 1b)、(5b)、(7b)、(9b);(1b)、(5b)、(7b)、(10b);(1b)、(5b)、(8b)、(9b);( 1b)、(5b)、(8b)、(10b);(1b)、(5b)、(9b)、(10b);(1b)、(6b)、(7b)、(8b);( 1b)、(6b)、(7b)、(9b);(1b)、(6b)、(7b)、(10b);(1b)、(6b)、(8b)、(9b);( 1b)、(6b)、(8b)、(10b);(1b)、(6b)、(9b)、(10b);(1b)、(7b)、(8b)、(9b);( 1b)、(7b)、(8b)、(10b);(1b)、(7b)、(9b)、(10b);(1b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b);(1b)、(2b)、(3b)、(4b)、(6b);(1b)、(2b)、(3 b)、(4b)、(7b);(1b)、(2b)、(3b)、(4b)、(8b);(1b)、(2b)、(3b)、(4b)、(9b );(1b)、(2b)、(3b)、(4b)、(10b);(1b)、(2b)、(3b)、(5b)、(6b);(1b)、(2b) ,(3b)、(5b)、(7b);(1b)、(2b)、(3b)、(5b)、(8b);(1b)、(2b)、(3b)、(5b)、(9b);(1b)、(2b)、(3b)、(5b)、(10b);(1b)、(2b)、(3b)、(6b)、(7b);(1b)、(2 b)、(3b)、(6b)、(8b);(1b)、(2b)、(3b)、(6b)、(9b);(1b)、(2b)、(3b)、(6b )、(10b);(1b)、(2b)、(3b)、(7b)、(8b);(1b)、(2b)、(3b)、(7b)、(9b);(1b) ,(2b)、(3b)、(7b)、(10b);(1b)、(2b)、(3b)、(8b)、(9b);(1b)、(2b)、(3b)、(8b)、(10b);(1b)、(2b)、(3b)、(9b)、(10b);(1b)、(2b)、(4b)、(5b)、(6b);(1b)、(2b)、(4b)、(5b)、(7b);(1b)、(2b)、(4b)、(5b)、(8b);(1b)、(2b)、( 4b)、(5b)、(9b);(1b)、(2b)、(4b)、(5b)、(10b);(1b)、(2b)、(4b)、(6b)、(7 b);(1b)、(2b)、(4b)、(6b)、(8b);(1b)、(2b)、(4b)、(6b)、(9b);(1b)、(2b) 、(4b)、(6b)、(10b);(1b)、(2b)、(4b)、(7b)、(8b);(1b)、(2b)、(4b)、(7b)、 (9b);(1b)、(2b)、(4b)、(7b)、(10b);(1b)、(2b)、(4b)、(8b)、(9b);(1b)、(2b)、(4b)、(8b)、(10b);(1b)、(2b)、(4b)、(9b)、(10b);(1b)、(2b)、(5b)、( 6b)、(7b);(1b)、(2b)、(5b)、(6b)、(8b);(1b)、(2b)、(5b)、(6b)、(9b);(1b )、(2b)、(5b)、(6b)、(10b);(1b)、(2b)、(5b)、(7b)、(8b);(1b)、(2b)、(5b) 、(7b)、(9b);(1b)、(2b)、(5b)、(7b)、(10b);(1b)、(2b)、(5b)、(8b)、(9b); (1b)、(2b)、(5b)、(8b)、(10b);(1b)、(2b)、(5b)、(9b)、(10b);(1b)、(2b)、 (6b)、(7b)、(8b);(1b)、(2b)、(6b)、(7b)、(9b);(1b)、(2b)、(6b)、(7b)、(1 0b);(1b)、(2b)、(6b)、(8b)、(9b);(1b)、(2b)、(6b)、(8b)、(10b);(1b)、(2 b)、(6b)、(9b)、(10b);(1b)、(2b)、(7b)、(8b)、(9b);(1b)、(2b)、(7b)、(8b )、(10b);(1b)、(2b)、(7b)、(9b)、(10b);(1b)、(2b)、(8b)、(9b)、(10b);(1 b)、(3b)、(4b)、(5b)、(6b);(1b)、(3b)、(4b)、(5b)、(7b);(1b)、(3b)、(4b)、(5b)、(8b);(1b)、(3b)、(4b)、(5b)、(9b);(1b)、(3b)、(4b)、(5b)、(10b);(1b)、(3b)、(4b)、(6b)、(7b);(1b)、(3b)、(4b)、(6b)、(8b);(1b)、(3b)、(4b)、(6b)、(9b);(1b)、(3b)、(4b)、(6b)、(10b);(1b)、(3b)、(4b)、(7b)、(8b );(1b)、(3b)、(4b)、(7b)、(9b);(1b)、(3b)、(4b)、(7b)、(10b);(1b)、(3b)、(4b)、(8b)、(9b);(1b)、(3b)、(4b)、(8b)、(10b);(1b)、(3b)、(4b)、(9b) (10b);(1b)、(3b)、(5b)、(6b)、(7b);(1b)、(3b)、(5b)、(6b)、(8b);(1b)、(3b)、(5b)、(6b)、(9b);(1b)、(3b)、(5b)、(6b)、(10b);(1b)、(3b)、(5b)、(7 b)、(8b);(1b)、(3b)、(5b)、(7b)、(9b);(1b)、(3b)、(5b)、(7b)、(10b);(1b)、(3b)、(5b)、(8b)、(9b);(1b)、(3b)、(5b)、(8b)、(10b);(1b)、(3b)、(5b) ,(9b)、(10b);(1b)、(3b)、(6b)、(7b)、(8b);(1b)、(3b)、(6b)、(7b)、(9b);(1b)、(3b)、(6b)、(7b)、(10b);(1b)、(3b)、(6b)、(8b)、(9b);(1b)、(3b)、( 6b)、(8b)、(10b);(1b)、(3b)、(6b)、(9b)、(10b);(1b)、(3b)、(7b)、(8b)、(9b);(1b)、(3b)、(7b)、(8b)、(10b);(1b)、(3b)、(7b)、(9b)、(10b);(1b)、( 3b), (8b), (9b), (10b);(1b), (4b), (5b), (6b), (7b);(1b), (4b), (5b), (6b), (8b);(1b), (4b), (5b), (6b), (9b);(1b), (4b), (5b), (6b), (10b);(1b )、(4b)、(5b)、(7b)、(8b);(1b)、(4b)、(5b)、(7b)、(9b);(1b)、(4b)、(5b)、(7b)、(10b);(1b)、(4b)、(5b)、(8b)、(9b);(1b)、(4b)、(5b)、(8b)、(10b);(1b)、(4b)、(5b)、(9b)、(10b);(1b)、(4b)、(6b)、(7b)、(8b);(1b)、(4b)、(6b)、(7b)、(9b);(1b)、(4b)、(6b)、(7b)、(10b);(1b)、(4b)、(6b)、(8b)、(9 b);(1b)、(4b)、(6b)、(8b)、(10b);(1b)、(4b)、(6b)、(9b)、(10b);(1b)、(4b)、(7b)、(8b)、(9b);(1b)、(4b)、(7b)、(8b)、(10b);(1b)、(4b)、(7b)、(9b) ,(10b);(1b)、(4b)、(8b)、(9b)、(10b);(1b)、(5b)、(6b)、(7b)、(8b);(1b)、(5b)、(6b)、(7b)、(9b);(1b)、(5b)、(6b)、(7b)、(10b);(1b)、(5b)、(6b)、( 8b)、(9b);(1b)、(5b)、(6b)、(8b)、(10b);(1b)、(5b)、(6b)、(9b)、(10b);( 1b)、(5b)、(7b)、(8b)、(9b);(1b)、(5b)、(7b)、(8b)、(10b);(1b)、(5b)、(7b )、(9b)、(10b);(1b)、(5b)、(8b)、(9b)、(10b);(1b)、(6b)、(7b)、(8b)、(9b );(1b)、(6b)、(7b)、(8b)、(10b);(1b)、(6b)、(7b)、(9b)、(10b);(1b)、(6b) ,(8b)、(9b)、(10b);(1b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b);(1b)、(2b)、(3b)、(4b)、(5b)、(7b);(1b)、(2b)、(3b)、(4b)、(5 b)、(8b);(1b)、(2b)、(3b)、(4b)、(5b)、(9b);(1b)、(2b)、(3b)、(4b)、(5b) 、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b);(1b)、(2b)、(3b)、(4b)、(6b)、( 8b);(1b)、(2b)、(3b)、(4b)、(6b)、(9b);(1b)、(2b)、(3b)、(4b)、(6b)、(10b);(1b)、(2b)、(3b)、(4b)、(7b)、(8b);(1b)、(2b)、(3b)、(4b)、(7b)、(9b);(1b)、(2b)、(3b)、(4b)、(7b)、(10b);(1b)、(2b)、(3b)、(4b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b)、(8b)、(10b);(1b)、(2b)、(3b)、(4b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(6b)、(7b);(1b)、(2b)、(3b)、(5b)、(6b)、(8b);(1b)、(2b)、(3b)、(5b)、(6b)、(9b);(1b)、(2b)、(3b)、(5b)、(6b)、(10b) ;(1b)、(2b)、(3b)、(5b)、(7b)、(8b);(1b)、(2b)、(3b)、(5b)、(7b)、(9b);(1b)、(2b)、(3b)、(5b)、(7b)、(10b);(1b)、(2b)、(3b)、(5b)、(8b)、(9b);(1b)、(2b)、(3b)、(5b)、(8b)、(10b);(1b)、(2b)、(3b)、(5b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(9b)、(10b);(1b)、(2b)、(3b)、(6b)、(7b)、(8b);(1b)、(2b)、(3b)、(6b)、(7b)、(8b);(1b)、(2b)、(3b)、(6b)、(7b)、(9b); (1b)、(2b)、(3b)、(6b)、(7b)、(10b);(1b)、(2b)、(3b)、(6b)、(8b)、(9b);(1b)、(2b)、(3b)、(6b)、(8b)、(10b);(1b)、(2b)、(3b)、(6b)、(9b)、(10b);(1b)、(2b)、(3b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(7b)、(8b)、(10b);(1b)、(2b)、(3b)、(7b)、(8b)、(10b);(1b)、(2b)、(3b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(8b)、(9b)、(10b) );(1b)、(2b)、(4b)、(5b)、(6b)、(7b);(1b)、(2b)、(4b)、(5b)、(6b)、(8b);(1b)、(2b)、(4b)、(5b)、(6b)、(9b);(1b)、(2b)、(4b)、(5b)、(6b)、(10b) ;(1b)、(2b)、(4b)、(5b)、(7b)、(8b);(1b)、(2b)、(4b)、(5b)、(7b)、(9b);(1b)、(2b)、(4b)、(5b)、(7b)、(10b);(1b)、(2b)、(4b)、(5b)、(8b)、(9b);(1b)、(2b)、(4b)、(5b)、(8b)、(10b);(1b)、(2b)、(4b)、(5b)、(9b)、(10b);(1b)、(2b)、(4b)、(6b)、(7b)、(8b);(1b)、(2b)、(4b)、(6b)、(7b)、(9b);(1b)、(2b)、(4b)、(6b)、(7b)、(10b);(1b)、(2b)、(4b)、(6b)、(8b)、(9b);(1b)、(2b)、(4b)、(6b)、(8b)、(10b);(1b)、(2b)、(4b)、(6b)、(9b)、(10b) ;(1b)、(2b)、(4b)、(7b)、(8b)、(9b);(1b)、(2b)、(4b)、(7b)、(8b)、(10b);(1b)、(2b)、(4b)、(7b)、(9b)、(10b);(1b)、(2b)、(4b)、(8b)、(9b)、(10b );(1b)、(2b)、(5b)、(6b)、(7b)、(8b);(1b)、(2b)、(5b)、(6b)、(7b)、(9b) ;(1b)、(2b)、(5b)、(6b)、(7b)、(10b);(1b)、(2b)、(5b)、(6b)、(8b)、(9b) ;(1b)、(2b)、(5b)、(6b)、(8b)、(10b);(1b)、(2b)、(5b)、(6b)、(9b)、(10b );(1b)、(2b)、(5b)、(7b)、(8b)、(9b);(1b)、(2b)、(5b)、(7b)、(8b)、(10b) ;(1b)、(2b)、(5b)、(7b)、(9b)、(10b);(1b)、(2b)、(5b)、(8b)、(9b)、(10b );(1b)、(2b)、(6b)、(7b)、(8b)、(9b);(1b)、(2b)、(6b)、(7b)、(8b)、(10b) ;(1b)、(2b)、(6b)、(7b)、(9b)、(10b);(1b)、(2b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(7b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(6b)、(7b );(1b)、(3b)、(4b)、(5b)、(6b)、(8b);(1b)、(3b)、(4b)、(5b)、(6b)、(9b);(1b)、(3b)、(4b)、(5b)、(6b)、(10b);(1b)、(3b)、(4b)、(5b)、(7b)、(8b);(1b)、(3b)、(4b)、(5b)、(7b)、(9b);(1b)、(3b)、(4b)、(5b)、(7b)、(10b);(1b)、(3b)、(4b)、(5b)、(8b)、(9b);(1b)、(3b)、(4b)、(5b)、(8b)、(10b);(1b)、(3b)、(4b)、(5b)、(9b)、(10b);(1b)、(3b)、(4b)、(6b)、(7b)、(8b);(1b)、(3b)、(4b)、(6b)、(7b)、(9b);(1b)、(3b)、(4b)、(6b)、(7b)、(10b); (1b)、(3b)、(4b)、(6b)、(8b)、(9b);(1b)、(3b)、(4b)、(6b)、(8b)、(10b);(1b)、(3b)、(4b)、(6b)、(9b)、(10b);(1b)、(3b)、(4b)、(7b)、(8b)、(9b);(1b)、(3b)、(4b)、(7b)、(8b)、(10b);(1b)、(3b)、(4b)、(7b)、(9b)、(10b);(1b)、(3b)、(4b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(8b)、(9b)、(10b);(1b)、(3b)、(5b)、(6b)、(7b)、(8b); (1b)、(3b)、(5b)、(6b)、(7b)、(9b);(1b)、(3b)、(5b)、(6b)、(7b)、(10b);(1b)、(3b)、(5b)、(6b)、(8b)、(9b);(1b)、(3b)、(5b)、(6b)、(8b)、(10b);(1b)、(3b)、(5b)、(6b)、(9b)、(10b);(1b)、(3b)、(5b)、(7b)、(8b)、(9b);(1b)、(3b)、(5b)、(7b)、(8b)、(10b);(1b)、(3b)、(5b)、(7b)、(9b)、(10b); (1b)、(3b)、(5b)、(8b)、(9b)、(10b);(1b)、(3b)、(6b)、(7b)、(8b)、(9b);(1b)、(3b)、(6b)、(7b)、(8b)、(10b);(1b)、(3b)、(6b)、(7b)、(9b)、(10b) ;(1b)、(3b)、(6b)、(8b)、(9b)、(10b);(1b)、(3b)、(7b)、(8b)、(9b)、(10b);(1b)、(4b)、(5b)、(6b)、(7b)、(8b);(1b)、(4b)、(5b)、(6b)、(7b)、(9b);(1b)、(4b)、(5b)、(6b)、(7b)、(10b);(1b)、(4b)、(5b)、(6b)、(8b)、(9b);(1b)、(4b)、(5b)、(6b)、(8b)、(10b);(1b)、(4b)、(5b)、(6b)、(9b)、(10b);(1b)、(4b)、(5b)、(7b)、(8b)、(9b);(1b)、(4b)、(5b)、(7b)、(8b)、(10b);(1b)、(4b)、(5b)、(7b)、(8b)、(10b);(1b)、(4b)、(5b)、(7b)、(9b)、(10b);(1b)、(4b)、(5b)、(8b)、(9b)、(10 b);(1b)、(4b)、(6b)、(7b)、(8b)、(9b);(1b)、(4b)、(6b)、(7b)、(8b)、(10b);(1b)、(4b)、(6b)、(7b)、(9b)、(10b);(1b)、(4b)、(6b)、(8b)、(9b)、(10b);(1b)、(4b)、(7b)、(8b)、(9b)、(10b);(1b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(5b)、(6b)、(7b)、(9b)、( 10b);(1b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(6b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(7b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(8b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(9b);(1b) ,(2b)、(3b)、(4b)、(5b)、(7b)、(8b);(1b)、(2b)、(3b)、(4b)、(5b)、(7b)、(9b);(1b)、(2b)、(3b)、(4b)、(5b)、(7b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b)、(5b)、(8b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b)、(8b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b)、(9b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b) )、(6b)、(8b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b)、(7b)、(8b)、(10b); (1b)、(2b)、(3b)、(4b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(6b)、(7b)、(8b);(1b)、(2b)、(3b)、(5 b)、(6b)、(7b)、(9b);(1b)、(2b)、(3b)、(5b)、(6b)、(7b)、(10b);(1b)、(2b )、(3b)、(5b)、(6b)、(8b)、(9b);(1b)、(2b)、(3b)、(5b)、(6b)、(8b)、(10b); (1b)、(2b)、(3b)、(5b)、(6b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(5b)、(7b)、(8b)、(10b);(1b)、(2b)、(3b)、(5 b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(8b)、(9b)、(10b);(1b)、(2 b)、(3b)、(6b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(6b)、(7b)、(8b)、(10b) ;(1b)、(2b)、(3b)、(6b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(4b)、( 5b), (6b), (7b), (8b);(1b), (2b), (4b), (5b), (6b), (7b), (9b);(1b), (2b), (4b), (5b), (6b), (7b), (10b);(1b), (2b), (4b), (5b), (6b), (8b), (9b);(1b)、(2b)、(4b)、(5b)、(6b)、(8b)、(10b);(1b)、(2b)、(4b)、(5b)、(6b)、(9b)、(10b);(1b)、(2b)、(4b)、(5b)、(7b)、(8b)、(9b);(1b)、(2b)、(4b)、(5b) )、(7b)、(8b)、(10b);(1b)、(2b)、(4b)、(5b)、(7b)、(9b)、(10b);(1b)、(2b)、(4b)、(5b)、(8b)、(9b)、(10b);(1b)、(2b)、(4b)、(6b)、(7b)、(8b)、(9b); (1b)、(2b)、(4b)、(6b)、(7b)、(8b)、(10b);(1b)、(2b)、(4b)、(6b)、(7b)、(9b)、(10b);(1b)、(2b)、(4b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(4b)、(7 b)、(8b)、(9b)、(10b);(1b)、(2b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(2b )、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(2b)、(5b)、(6b)、(7b)、(9b)、(10b) ;(1b)、(2b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(6b)、(7b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、( 5b), (6b), (7b), (8b);(1b), (3b), (4b), (5b), (6b), (7b), (9b);(1b), (3b), (4b), (5b), (6b), (7b), (10b);(1b), (3b), (4b), (5b), (6b), (8b), (9b); (1b)、(3b)、(4b)、(5b)、(6b)、(8b)、(10b);(1b)、(3b)、(4b)、(5b)、(6b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(7b)、(8b)、(9b);(1b)、(3b)、(4b)、(5b) )、(7b)、(8b)、(10b);(1b)、(3b)、(4b)、(5b)、(7b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(6b)、(7b)、(8b)、(9b);(1b)、(3b)、(4b)、(6b)、(7b)、(8b)、(10b);(1b)、(3b)、(4b)、(6b)、(7b)、(9b)、(10b);(1b)、(3b)、(4b)、(6b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(7b)、(8b)、(9b)、(10b);(1b)、(3b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(3b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(3b)、(5b)、(6b)、(7b)、(9b) )、(10b);(1b)、(3b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(3b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(3b)、(6b)、(7b)、(8b)、(9b)、(10b);(1b)、(4b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(4b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(4b)、(5b)、(6b)、(7b)、(9b)、(10b);(1b)、(4b)、(5b)、(6b)、(7b)、(9b)、(10b);(1b)、(4b)、(5b)、(6b)、(7b)、(9b)、(10b);(1b)、(4b)、(5b)、(6b)、(7b)、(9b)、( 8b)、(9b)、(10b);(1b)、(4b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(4b)、(6b)、(7b)、(8b)、(9b)、(10b);(1b)、(5b)、(6b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(7b)、(8b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(7b)、(9b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(7b)、(10b); (1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(8b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(6b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b)、(5b)、(7b)、(8b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(5b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b)、(8b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(4b)、(7b)、(8b)、(9b)、(10b); (1b)、(2b)、(3b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(2b)、(3b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(2b)、(3b)、(5b)、(6b)、(7b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(3b)、(5b)、(7b)、(9b)、(10b); (1b)、(2b)、(4b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(2b)、(4b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(2b)、(4b)、(5b)、(6b)、(7b)、(9b)、(10b);(1b)、(2b)、(4b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(2b)、(4b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(4b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b)、(2b)、(4b)、(6b)、(7b)、(9b)、(10b); (1b)、(2b)、(5b)、(6b)、(7b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(6b)、(7b)、(8b)、(9b);(1b)、(3b)、(4b)、(5b)、(6b)、(7b)、(8b)、(10b);(1b)、(3b)、(4b)、(5b)、(6b)、(7b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(6b)、(8b)、(9b)、(10b);(1b)、(3b)、(4b)、(5b)、(7b)、(8b)、(9b)、(10b);(1b), (3b), (4b), (6b), (7b), (8b), (9b), (10b); (1b), (3b), (5b), (6b), (7b), (8b), (9 b), (10b); (1b), (4b), (5b), (6b), (7b), (8b), (9b), (10b); (1b), (2b), (3b), (4b), (5b) ), (6b), (7b), (8b), (9b); (1b), (2b), (3b), (4b), (5b), (6b), (7b), (8b), (10b); (1b), (2b), (3b), (4b), (5b), (6b), (7b), (9b), (10b); (1b), (2b), (3b), (4b), (5b), (6b), (8b) ), (9b), (10b); (1b), (2b), (3b), (4b), (5b), (7b), (8b), (9b), (10b); (1b), (2b), (3b) , (4b), (6b), (7b), (8b), (9b), (10b); (1b), (2b), (3b), (5b), (6b), (7b), (8b), (9b), (1 0b); (1b), (2b), (4b), (5b), (6b), (7b), (8b), (9b), (10b); (1b), (3b), (4b), (5b), (6b) ), (7b), (8b), (9b), (10b); (1b), (2b), (3b), (4b), (5b), (6b), (7b), (8b), (9b), (10b). ;

[0389] In the combinations described above, “(1b)” is independently selected from (1b)(i), (1b)(ii), (1b)(iii), and (1b)(iv); and “(2b)” is independently selected from (2b)(i), (2b)(ii), (2b)(iii), and (2b)(iv) above; and “(3b)” is independently selected from (3b)(i), (3b)(ii), (3b)(iii), and (3b)(iv) above; and “(4b)” is independently selected from (4b) above. (i), (4b)(ii), (4b)(iii) and (4b)(iv); and “(5b)” is independently selected from (5b)(i), (5b)(ii), (5b)(iii) and (5b)(iv) above; and “(6b)” is independently selected from (6b)(i) and (6b)(ii) above; and “(7b)” is independently selected from (7b)(i) and (7b)(ii) above; and “(8b)” is independently selected from (8b)(i) and (8b)(ii) above.

[0390] All possible combinations of the features described above are included. For illustrative purposes only, cancers containing the feature combinations “(1b), (2b)” specifically cover cancers containing the following features: (1b)(i) and (2b)(i); (1b)(i) and (2b)(ii); (1b)(i) and (2b)(iii); (1b)(i) and (2b)(iv); (1b)(ii) and (2b)(i); (1b)(ii) and (2b)(ii); (1b)(ii) and (2b)(iii); (1b)(ii) and (2b)(iii); (1 b)(ii) and (2b)(iv); (1b)(iii) and (2b)(i); (1b)(iii) and (2b)(ii); (1b)(iii) and (2b)(iii); (1b)(iii) and (2b)(iv); (1b)(iv) and (2b)(i); (1b)(iv) and (2b)(ii); (1b)(iv) and (2b)(iii); and (1b)(iv) and (2b)(iv).

[0391] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist is homozygous for activating mutations in genes encoding positive regulators of HER3-mediated signal transduction, selected from the following: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer is a heterozygote for an activating mutation of a gene encoding a HER3-mediated positive regulator of signal transduction, selected from the following: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer includes genetic variations that lead to increased expression of genes or enhanced activity of gene products encoding HER3-mediated positive regulators of signal transduction selected from the following: PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5.

[0392] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist is a homozygous gene for activating mutations of the following genes encoding positive regulators of the MAPK / ERK pathway: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1. In some embodiments, the cancer is a heterozygote for activating mutations in genes encoding positive signaling regulators of the MAPK / ERK pathway, selected from the following: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1. In some embodiments, the cancer includes genetic variations selected from the following that result in increased expression of genes encoding positive regulators of the MAPK / ERK pathway signaling pathway or enhanced activity of gene products: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1.

[0393] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented by using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist is a homozygous gene for activating mutations of the following genes encoding positive regulators of the PI3K / AKT / mTOR pathway: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer is a heterozygote of an activating mutation in a gene selected from the following gene encoding a positive regulator of the PI3K / AKT / mTOR pathway: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5. In some embodiments, the cancer includes genetic variations selected from the following that lead to increased expression of genes encoding positive signaling regulators of the PI3K / AKT / mTOR pathway or enhanced activity of gene products: ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CB, PIK3CD, mTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5.

[0394] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist is a homozygous individual with an inactivating mutation in a gene encoding a negative regulator of HER3-mediated signal transduction selected from the following: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the cancer is a heterozygous individual with an inactivating mutation in a gene encoding a negative regulator of HER3-mediated signal transduction selected from the following: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the cancer comprises a genetic variant selected from the following that results in reduced expression of a gene encoding a negative regulator of HER3-mediated signal transduction or reduced activity of its gene product: PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0395] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist is homozygous for an NF1 gene inactivation mutation. In some embodiments, the cancer of a heterozygous inactivation mutation is heterozygous for an NF1 gene inactivation mutation. In some embodiments, the cancer containing genetic variations that result in reduced expression or decreased activity of the gene product is homozygous for an NF1 inactivation mutation.

[0396] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist is a homozygous individual with an inactivating mutation in a gene encoding a negative regulator of the PI3K / AKT / mTOR pathway, selected from the following: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1. In some embodiments, the cancer is a heterozygous individual with an inactivating mutation in a gene encoding a negative regulator of the PI3K / AKT / mTOR pathway, selected from the following: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1. In some embodiments, the cancer includes a genetic variant selected from the following that results in reduced expression of a gene encoding a negative regulator of the PI3K / AKT / mTOR pathway or reduced activity of its gene product: PPP2CA, PIK3R1, PIK3R2, BAD, and PHLPP1.

[0397] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist (i) comprises a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) comprises a genetic variant that results in increased KRAS expression or enhanced gene product activity. In some embodiments, the cancer (i) comprises MET amplification, and (ii) comprises a KRAS activating mutation.

[0398] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in increased PIK3CA expression or enhanced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains a PIK3CA activating mutation.

[0399] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist includes: (i) a genetic variant leading to increased MET expression or enhanced gene product activity, and (ii) a genetic variant leading to increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer (i) includes MET amplification, and (ii) includes a BRAF activating mutation.

[0400] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains PTEN inactivation mutations.

[0401] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in increased KRAS expression or enhanced gene product activity, and (iii) contains a genetic variant that results in increased PIK3CA expression or enhanced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains a KRAS activating mutation, and (iii) contains a PIK3CA activating mutation.

[0402] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in increased KRAS expression or enhanced gene product activity, and (iii) contains a genetic variant that results in increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains a KRAS activating mutation, and (iii) contains a BRAF activating mutation.

[0403] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in increased KRAS expression or enhanced gene product activity, and (iii) contains a genetic variant that results in decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains a KRAS activating mutation, and (iii) contains a PTEN inactivating mutation.

[0404] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in increased PIK3CA expression or enhanced gene product activity, and (iii) contains a genetic variant that results in increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains a PIK3CA activating mutation, and (iii) contains a BRAF activating mutation.

[0405] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, (ii) contains a genetic variant that results in increased PIK3CA expression or enhanced gene product activity, and (iii) contains a genetic variant that results in decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, (ii) contains a PIK3CA activating mutation, and (iii) contains a PTEN inactivating mutation.

[0406] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity, and (ii) contains a genetic variant that results in increased BRAF expression or enhanced gene product activity, and (iii) contains a genetic variant that results in decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) contains MET amplification, and (ii) contains a BRAF activating mutation, and (iii) contains a PTEN inactivating mutation.

[0407] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist includes: (i) a genetic variant leading to increased MET expression or enhanced gene product activity; (ii) a genetic variant leading to increased KRAS expression or enhanced gene product activity; (iii) a genetic variant leading to increased PIK3CA expression or enhanced gene product activity; and (iv) a genetic variant leading to increased BRAF expression or enhanced gene product activity. In some embodiments, the cancer includes: (i) MET amplification; (ii) KRAS activating mutation; (iii) PIK3CA activating mutation; and (iv) BRAF activating mutation.

[0408] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist includes: (i) a genetic variant leading to increased MET expression or enhanced gene product activity; (ii) a genetic variant leading to increased KRAS expression or enhanced gene product activity; (iii) a genetic variant leading to increased PIK3CA expression or enhanced gene product activity; and (iv) a genetic variant leading to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer includes: (i) MET amplification; (ii) KRAS activating mutation; (iii) PIK3CA activating mutation; and (iv) PTEN inactivating mutation.

[0409] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist: (i) contains a genetic variant that results in increased MET expression or enhanced gene product activity; (ii) contains a genetic variant that results in increased KRAS expression or enhanced gene product activity; (iii) contains a genetic variant that results in increased BRAF expression or enhanced gene product activity; and (iv) contains a genetic variant that results in decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer: (i) contains MET amplification; (ii) contains a KRAS activating mutation; (iii) contains a BRAF activating mutation; and (iv) contains a PTEN inactivating mutation.

[0410] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist includes: (i) a genetic variant leading to increased MET expression or enhanced gene product activity; (ii) a genetic variant leading to increased PIK3CA expression or enhanced gene product activity; (iii) a genetic variant leading to increased BRAF expression or enhanced gene product activity; and (iv) a genetic variant leading to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer includes: (i) MET amplification; (ii) PIK3CA activating mutation; (iii) BRAF activating mutation; and (iv) PTEN inactivating mutation.

[0411] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented using an antigen-binding molecule that binds to HER3 in combination with a HER3-mediated signal transduction antagonist includes: (i) a genetic variant leading to increased MET expression or enhanced gene product activity; (ii) a genetic variant leading to increased KRAS expression or enhanced gene product activity; (iii) a genetic variant leading to increased PIK3CA expression or enhanced gene product activity; (iv) a genetic variant leading to increased BRAF expression or enhanced gene product activity; and (v) a genetic variant leading to decreased PTEN expression or reduced gene product activity. In some embodiments, the cancer includes: (i) MET amplification; (ii) KRAS activating mutation; (iii) PIK3CA activating mutation; (iv) BRAF activating mutation; and (v) PTEN inactivating mutation.

[0412] Cancer characterized by the presence of genetic variations that lead to increased expression of genes on the long arm of chromosome 3 or enhanced activity of gene products.

[0413] The various aspects and embodiments described in this disclosure relate to cancers comprising genetic variations that result in increased expression of genes on the long arm of chromosome 3 or enhanced activity of gene products. It should be understood that the cancers described in this section may be further characterized according to the features described in the preceding “Cancer” section.

[0414] Early-stage squamous cell carcinoma is characterized by amplification of the long arm of chromosome 3 (Chr3q), a known genetic aberration associated with exposure to carcinogens (Rooney et al., *Oncologist*, 2013, Vol. 18, No. 6, pp. 707-716). Chr3q amplification leads to enhanced transcriptional activity of several oncogenes, including TP63 (which directly promotes the expression of the HER3 ligand NRG1), SOX2 (which directly promotes the expression of the EGFR ligand), and the PIK3CA gene (which increases PI3K pathway activation) (Perez-Moreno et al., *Clinical Cancer Research*, 2012, Vol. 8, No. 9, pp. 2443-2451).

[0415] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure is characterized by one or more of the following features:

[0416] (6a)(i) includes genetic variations that lead to increased TP63 expression or enhanced activity of the gene product.

[0417] (6a)(ii) includes TP63 amplification (TP63 amplification as described herein).

[0418] (7a)(i) contains genetic variations that lead to increased PIK3CA expression or enhanced activity of the gene product.

[0419] (7a)(ii) includes PIK3CA amplification (PIK3CA amplification as described herein).

[0420] (8a)(i) includes genetic variations that lead to increased SOX2 expression or enhanced activity of gene products.

[0421] (8a)(ii) includes SOX2 amplification (the SOX2 amplification described herein).

[0422] In some implementation examples, the cancer to be treated / prevented, as described in this disclosure, can be characterized by one of the following combinations of features (refer to the preceding paragraph): (6a); (7a); (8a); (6a), (7a); (6a), (8a); (7a), (8a); (6a), (7a), (8a).

[0423] In the combinations described above, “(6a)” is independently selected from (6a)(i) and (6a)(ii) above; “(7a)” is independently selected from (7a)(i) and (7a)(ii) above; and “(8a)” is independently selected from (8a)(i) and (8a)(ii) above.

[0424] All possible combinations of the features described above are included. For illustrative purposes only, cancers containing the feature combination “(6a),(7a)” specifically cover cancers containing the following features: (6a)(i) and (7a)(i); (6a)(i) and (7a)(ii); (6a)(ii) and (7a)(i); and (6a)(ii) and (7a)(ii).

[0425] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes an amplification of the long arm of chromosome 3.

[0426] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes genetic variations that result in increased expression or enhanced activity of gene products selected from the following: ADIPOQ, AMOTL2, ARHGAP31, TIMMDC1, C3orf70, CAMPD1, CCDC80, CD200R1, CHST13, CLDND1, CPN2, CPOX, DPPA2, DTX3L, DZIP3, EAF2, EFCC1, ETM1, ETV5, FAM3D, FAM43A, FAM162A, FBXO40, FILIP1L, GYG1, HACD2, HGD, IFT122, K The following genes are listed: IAA1257, LINC01279, LNCR5, LMLN, LRRC15, LSG1, MB21D2, MCCC1, MORC1, MYLK, NEPRO, NFKBIZ, OTOL1, PARP14, PCCB, PDCD10, PIK3CA, PISRT1, PROSER1, RAB7, RASA2, RETNLB, RHO, RIOX2, SELT, SENP7, SERP1, SOX2, SOX2OT, SPG14, SRPRB, TEX55, TMEM44, TM4SF1, TMPRSS7, TP63, TRAT1, USH3A, ZBED2, and ZNF9. In some embodiments, the cancer includes genetic variations that result in increased expression or enhanced activity of gene products selected from the following genes: TP63, PIK3CA, and SOX2.

[0427] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure comprises amplifications of genes selected from: ADIPOQ, AMOTL2, ARHGAP31, TIMMDC1, C3orf70, CAMPD1, CCDC80, CD200R1, CHST13, CLDND1, CPN2, CPOX, DPPA2, DTX3L, DZIP3, EAF2, EFCC1, ETM1, ETV5, FAM3D, FAM43A, FAM162A, FBXO40, FILIP1L, GYG1, HACD2, HGD, IFT122, KIAA1257, L The gene sequences selected from INC01279, LNCR5, LMLN, LRRC15, LSG1, MB21D2, MCCC1, MORC1, MYLK, NEPRO, NFKBIZ, OTOL1, PARP14, PCCB, PDCD10, PIK3CA, PISRT1, PROSER1, RAB7, RASA2, RETNLB, RHO, RIOX2, SELT, SENP7, SERP1, SOX2, SOX2OT, SPG14, SRPRB, TEX55, TMEM44, TM4SF1, TMPRSS7, TP63, TRAT1, USH3A, ZBED2, and ZNF9 are included. In some embodiments, the cancer comprises amplifications of genes selected from TP63, PIK3CA, and SOX2.

[0428] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes genetic variations that lead to increased TP63 expression or enhanced activity of the gene product. In some embodiments, the cancer includes TP63 amplification.

[0429] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented, as described in this disclosure, includes genetic variations that lead to increased PIK3CA expression or enhanced activity of the gene product. In some embodiments, the cancer includes PIK3CA amplification.

[0430] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes genetic variations that lead to increased SOX2 expression or enhanced activity of the gene product. In some embodiments, the cancer includes SOX2 amplification.

[0431] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure (i) includes genetic variants that lead to increased TP63 expression or enhanced gene product activity, and (ii) includes genetic variants that lead to increased PIK3CA expression or enhanced gene product activity. In some embodiments, the cancer (i) includes TP63 amplification, and (ii) includes PIK3CA amplification.

[0432] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure (i) includes genetic variants that result in increased TP63 expression or enhanced gene product activity, and (ii) includes genetic variants that result in increased SOX2 expression or enhanced gene product activity. In some embodiments, the cancer (i) includes TP63 amplification, and (ii) includes SOX2 amplification.

[0433] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure (i) includes genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (ii) includes genetic variants that lead to increased SOX2 expression or enhanced gene product activity. In some embodiments, the cancer (i) includes PIK3CA amplification, and (ii) includes SOX2 amplification.

[0434] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure (i) includes genetic variants that lead to increased TP63 expression or enhanced gene product activity, and (ii) includes genetic variants that lead to increased PIK3CA expression or enhanced gene product activity, and (iii) includes genetic variants that lead to increased SOX2 expression or enhanced gene product activity. In some embodiments, the cancer (i) includes TP63 amplification, and (ii) includes PIK3CA amplification, and (iii) includes SOX2 amplification.

[0435] Cancer characterized by the presence of genetic variations that lead to increased expression of genes on the short arm of chromosome 7 or enhanced activity of gene products.

[0436] The various aspects and embodiments described in this disclosure relate to cancers comprising genetic variations that result in increased expression of genes on the short arm of chromosome 7 or enhanced activity of gene products. It should be understood that the cancers described in this section may be further characterized according to the features described in the preceding section on “Cancer”.

[0437] Frequent amplification of the short arm (Chr7p) of chromosome 7 encoding EGFR was found in LUSC (Couceiro et al., Revista Portuguesa de Pneumologia, 2010, Vol. 16, No. 3, pp. 453-462).

[0438] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure is characterized by one or more of the following features:

[0439] (10a)(i) includes genetic variations that lead to increased EGFR expression or enhanced activity of gene products.

[0440] (10a)(ii) includes EGFR amplification (EGFR amplification as described herein).

[0441] In various aspects and embodiments as described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes an amplification of the short arm of chromosome 7.

[0442] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes genetic variations that lead to increased EGFR expression or enhanced activity of its gene product. In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes EGFR amplification.

[0443] Cancer characterized by the presence of genetic variations that lead to reduced gene expression or decreased gene product activity on the short arm of chromosome 3.

[0444] The various aspects and embodiments described in this disclosure relate to cancers comprising genetic variations that result in reduced expression of genes on the short arm of chromosome 3 or decreased activity of gene products. It should be understood that the cancers described in this section may be further characterized according to the features described in the preceding section on “Cancer”.

[0445] Early-stage squamous cell carcinoma is characterized by the deletion of the short arm of chromosome 3 (Chr3p), a known genetic aberration associated with exposure to carcinogens (Rooney et al., *Oncologist*, 2013, Vol. 18, No. 6, pp. 707-716). Chr3p deletion leads to the loss of several putative tumor suppressor proteins, including TUSC2, a tumor suppressor candidate protein that can inhibit EGFR (Perez-Moreno et al., *Clinical Cancer Research*, 2012, Vol. 8, No. 9, pp. 2443-2451). TUSC2 deletion (TCGA) has been found in approximately 12-17% of squamous cell carcinoma patients. Furthermore, studies have shown that TUSC2 can negatively regulate EGFR signaling (Dai et al., *PLoS One*, 2015, Vol. 10, No. 6, p. e0123967; Cao et al., *Scientific Reports*, 2016, Vol. 6, p. 35741).

[0446] In the various aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure is characterized by one or more of the following features:

[0447] (11a)(i) does not contain genetic variations that result in reduced TUSC2 expression or decreased gene product activity;

[0448] (11a)(ii) does not include TUSC2 deletion (the TUSC2 deletion described herein).

[0449] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes a deletion of the short arm of chromosome 3.

[0450] In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes genetic variations that result in reduced TUSC2 expression or decreased activity of the gene product. In all aspects and embodiments described in this disclosure, the cancer to be treated / prevented as described in this disclosure includes TUSC2 deletion.

[0451] antigen-binding molecules

[0452] This disclosure relates to the therapeutic and preventative uses of antigen-binding molecules that bind to HER3.

[0453] An "antigen-binding molecule" is a molecule capable of binding to a given target antigen. Antigen-binding molecules include antibodies (i.e., immunoglobulins (Igs)) and their antigen-binding fragments. As described herein, "antibody" includes monoclonal antibodies, polyclonal antibodies, monospecific and multispecific (e.g., bispecific, trispecific, etc.) antibodies, and antigen-binding molecules derived from antibodies, such as scFv, scFab, bispecific antibodies, trispecific antibodies, scFv-Fc, microantibodies, and single-domain antibodies (e.g., VhH, etc.). Antigen-binding fragments of antibodies include, for example, Fv, Fab, F(ab')2, and F(ab') fragments. In some embodiments, the antigen-binding molecule may be an antibody or its antigen-binding fragment.

[0454] The antigen-binding molecules described in this disclosure also include antibody-derived molecules, such as molecules comprising an antigen-binding region / domain derived from an antibody. Antibody-derived antigen-binding molecules may comprise an antigen-binding region / domain containing or composed of an antigen-binding region of an antibody (such as an antigen-binding fragment of an antibody). In some embodiments, the antigen-binding region / domain of the antibody-derived antigen-binding molecule may be or comprise an Fv (e.g., present as scFv) or Fab region of the antibody, or the entire antibody. For example, antigen-binding molecules described in this disclosure include antibody-drug conjugates (ADCs) comprising a (cytotoxic) drug portion (described below). The antigen-binding molecules described in this disclosure also include multispecific antigen-binding molecules, such as immune cell adaptor molecules containing a domain for recruiting (effective) immune cells (see Goebeler and Bargou, *Nature Reviews Clinical Oncology*, 2020, Vol. 17, pp. 418-434 and Ellerman, *Methods*, 2019, Vol. 154, pp. 102-117, both of which are incorporated herein by reference in their entirety), including BiTE, BiKE, and TriKE. The antigen-binding molecules described in this disclosure also include chimeric antigen receptors (CARs), which are recombinant receptors that simultaneously provide antigen binding and T cell activation functions (for the structure, function, and engineering of CARs, see Dotti et al., *Immunol Rev*, 2014, Vol. 257, No. 1 and Jayaraman et al., *EBioMedicine*, 2020, Vol. 58, pp. 102-931, both of which are incorporated herein by reference in their entirety).

[0455] The antigen-binding molecules described in this disclosure comprise one or more portions capable of binding to a target antigen. In some embodiments, the portion capable of binding to the target antigen comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) of an antibody capable of specifically binding to the target antigen. In some embodiments, the portion capable of binding to the target antigen comprises or is composed of an aptamer capable of binding to the target antigen, such as a nucleic acid aptamer (see Zhou and Rossi, *Nature Reviews Drug Discovery*, 2017, Vol. 16, No. 3, pp. 181-202). In some embodiments, the portion capable of binding the target antigen comprises or is composed of antigen-binding peptides / polypeptides, such as peptide aptamers, thioredoxins, monoclonal antibody mimics, anticalcin, Kunitz domains, avimer, cysteine ​​knot peptides, fynomers, atrimers, DARPin, affinity molecules, nanobodies (i.e., single-domain antibodies sdAbs), affilin, armadillo repeat protein (ArmRP), OBody, or fibronectin, see Reverdatto et al., Current Topics in Medicinal Chemistry (Curr Top Med Chem), 2015, Vol. 15, No. 12, pp. 1082-1101, which is incorporated herein by reference in its entirety (see also Boersma et al., J Biol Chem, 2011, Vol. 286, pp. 41273-85 and Emanuel et al., Monoclonal Antibodies (Mabs), 2011, Vol. 3, pp. 38-48).

[0456] As used herein, a "peptide" refers to a chain of two or more amino acid monomers linked by peptide bonds. Peptides typically have a length of about 2 to 50 amino acids. A "polypeptide" is a polymer chain composed of two or more peptides. Polypeptides typically have a length of more than about 50 amino acids.

[0457] As described in this disclosure, antigen-binding molecules typically comprise antigen-binding domains VH and VL containing antibodies capable of specifically binding to target antigens. The antigen-binding domain consisting of VH and VL is also referred to herein as the Fv region.

[0458] Antigen-binding molecules may be or contain antigen-binding peptides or antigen-binding peptide complexes. Antigen-binding molecules may contain multiple peptides that collectively constitute an antigen-binding domain. The peptides may be bound covalently or non-covalently. In some embodiments, the peptide constitutes a component of a larger-scale peptide comprising this peptide (e.g., scFv containing VH and VL, or scFab containing VH-CH1 and VL-CL).

[0459] Antigen-binding molecules can refer to non-covalent or covalent complexes of multiple polypeptides (such as 2, 3, 4, 6 or 8 polypeptides), such as IgG-like antigen-binding molecules containing two heavy chain polypeptides and two light chain polypeptides.

[0460] As described in this disclosure, antigen-binding molecules can be designed and prepared from sequences of monoclonal antibodies capable of binding to HER3. Antigen-binding regions of antibodies, such as single-chain variable fragments (scFv), Fab, and F(ab')2 fragments, can also be used / provided. An "antigen-binding region" is any antibody fragment capable of binding to the specific target of a given antibody.

[0461] Antibodies typically contain six complementarity-determining regions (CDRs): three in the heavy chain variable region (HC-CDR1, HC-CDR2, and HC-CDR3) and three in the light chain variable region (LC-CDR1, LC-CDR2, and LC-CDR3). These six CDRs together constitute the antibody determinant cluster, which is the portion of the antibody that binds to the target antigen.

[0462] The VH and VL regions contain frame areas (FRs) located on both sides of each CDR, which provide support for the CDR. From the N end to the C end, the VH region contains the following structure: N end - [HC-FR1] - [HC-CDR1] - [HC-FR2] - [HC-CDR2] - [HC-FR3] - [HC-CDR3] - [HC-FR4] - C end; the VL region contains the following structure: N end - [LC-FR1] - [LC-CDR1] - [LC-FR2] - [LC-CDR2] - [LC-FR3] - [LC-CDR3] - [LC-FR4] - C end.

[0463] There are currently several conventions for defining antibody CDR and FR, such as those described by Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991; Chothia et al., Journal of Molecular Biology, 1987, Vol. 196, pp. 901-917; and the VBASE2 system, described by Retter et al., Nucleic Acids Research, 2005, Vol. 33, pp. 671-674. The CDR and FR of the VH and VL regions of the antibody clones described herein are defined according to the international IMGT (Immunogenetics) information system (LeFranc et al., Nucleic Acids Res., 2015, Vol. 43, D413-22), which adopts the IMGT V-DOMAIN numbering rules described by Lefranc et al., Developmental and Comparative Immunology, 2003, Vol. 27, pp. 55-77. In a preferred embodiment, the CDR and FR of the antigen-binding molecule as described herein are defined according to the IMGT information system.

[0464] The VH and VL regions of the antigen-binding region of an antibody together constitute the Fv region. In some embodiments, the antigen-binding molecule as described in this disclosure comprises or consists of an Fv region that binds to HER3. In some embodiments, the VH and VL regions of the Fv region are linked by a linker peptide sequence to form a single polypeptide chain, i.e., a single-chain Fv (scFv).

[0465] The antigen-binding region of an antibody comprises the VL region and the light chain constant region (CL), as well as the VH region and the heavy chain constant 1 region (CH1), which together constitute the Fab region. In some embodiments, the antigen-binding molecule includes a Fab region containing VH, CH1, VL, and CL (such as Cκ or Cλ). In some embodiments, the Fab region includes a polypeptide containing VH and CH1 regions (such as a VH-CH1 fusion polypeptide) and a polypeptide containing VL and CL (such as a VL-CL fusion polypeptide). In some embodiments, the Fab region includes a polypeptide containing VH and CL regions (such as a VH-CL fusion polypeptide) and a polypeptide containing VL and CH1 (such as a VL-CH1 fusion polypeptide), i.e., in some embodiments, the Fab region is a CrossFab region. In some embodiments, the VH, CH1, VL, and CL of the Fab region or CrossFab region are linked by linker peptide regions to form a single polypeptide chain, i.e., a single-chain Fab (scFab) or a single-chain CrossFab (scCrossFab).

[0466] In some implementations, the antigen-binding molecules described herein comprise or consist of whole antibodies that bind HER3. As used herein, “whole antibody” refers to an antibody having a structure substantially similar to that of immunoglobulins (Ig). Different types of immunoglobulins and their structures are described by Schroeder and Cavacini, *J Allergy Clin Immunology*, 2010, Vol. 125, No. 202, pp. 41-52, which is incorporated herein by reference in its entirety.

[0467] Immunoglobulins G (IgG) are glycoproteins of approximately 150 kDa, consisting of two heavy chains and two light chains. From the N-terminus to the C-terminus, the heavy chain contains the VH domain followed by a constant region of the heavy chain containing three constant domains (CH1, CH2, and CH3). Similarly, the light chain contains the VL domain followed by the CL domain. Based on the heavy chain, immunoglobulins can be classified as IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM. The light chains can be κ or λ type.

[0468] In some implementations, the antigen-binding molecule comprises or consists of IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM that bind to HER3.

[0469] In some embodiments, the antigen-binding molecule includes a CDR region capable of binding to HER3. In some embodiments, the antigen-binding molecule includes an FR region capable of binding to HER3. In some embodiments, the antigen-binding molecule includes both a CDR region and an FR region capable of binding to HER3. That is, in some embodiments, the antigen-binding molecule includes a VH region and a VL region capable of binding to HER3.

[0470] In some embodiments, the antigen-binding molecule capable of binding HER3, as described in this disclosure, may be selected from: any of the embodiments described in WO2019185878A1 (which is incorporated herein by reference in its entirety), 10D1F (described in WO2019185878A1), seribantumab (also known as MM-121, described in Schoeberl et al., *Sci. Signal*, 2009, Vol. 2, No. 77, p. 31), elgemtumab (also known as LJM-716, described in Garner et al., *Cancer Research*, 2013, Vol. 73, pp. 6024-6035), and patritumab (also known as U-1287 and AMG-888, described in Shimizu et al., *Cancer Chemotherapy and Pharmacology*). Pharmacol.) 2017 Vol. 79 No. 3 pp. 489-495), GSK2849330 (described by Clarke et al., *European Journal of Cancer*, Vol. 50, pp. 98-99), lumretuzumab (also known as RG7116 and RO-5479599, described by Mirschberger et al., *Cancer Research*, Vol. 73 No. 16, pp. 5183-5194), CDX-3379 (also known as KTN3379, described by Lee et al., *Proceedings of the National Academy of Sciences of the United States of America*, Vol. 112 No. 43, pp. 13225), AV-203 (also known as CAN-017, described by Meetze et al., *European Journal of Cancer*, Vol. 79 No. 3, pp. 489-495), GSK2849330 (described by Clarke et al., *European Journal of Cancer*, Vol. 50, pp. 98-99), lumretuzumab (also known as RG7116 and RO-5479599, described by Mirschberger et al., *Cancer Research*, Vol. 73, pp. 16, pp. 5183-5194), CDX-3379 (also known as KTN3379, described by Lee et al., *Proceedings of the National Academy of Sciences of the United States of America*, Vol. 112, pp. 43, pp. 13225), AV-203 (also known as CAN-017, described by Meetze et al., *European Journal of Cancer*, Vol. 73, pp. 16, pp. 13225), AV-20 Cancer, 2012, Vol. 48, p. 126; barecetamab (also known as ISU104, described by Kim et al., Cancer Research, 2018, Vol. 78 (Supplement 13), Abstract #830); TK-A3 and TK-A4 (described by Malm et al., Monoclonal Antibodies, 2016, Vol. 8, pp. 1195-1209); MP-EV20 (described by Sala et al., Translational Oncology).(2013, Vol. 6, pp. 676-84), 1A5-3D4 (described by Wang et al., *Cancer Lett*, Vol. 380, 2016, pp. 20-30), 9F7-F11 and 16D3-C1 (described by Lazrek et al., *Neoplasia*, Vol. 15, 2013, pp. 335-47), NG33, A5 and F4 (described by Gaborit et al., *Proceedings of the National Academy of Sciences of the United States of America*, Vol. 112, 2015, pp. 839-44), huHER3-8 (described by Kugel et al., *Cancer Research*, Vol. 74, 2014, pp. 4122-32), REGN1400 (described by Zhang et al., *Molecular Cancer Therapy*). (Ther) 2014, Vol. 13, pp. 1345-1355) and zenocutuzumab (also known as MCLA-128, described in de Vries Schultink et al., *Clinical Pharmacokinetics*, Vol. 59, pp. 875-884, 2020).

[0471] In some embodiments, the antigen-binding molecule is selected from 10D1F and seribantumab. In some embodiments, the antigen-binding molecule is 10D1F.

[0472] In some embodiments, the antigen-binding molecule comprises a CDR region or VH and VL regions selected from the following HER3-binding antibody clones: 10D1_c89, 10D1, 10D1_c75, 10D1_c76, 10D1_c77, 10D1_c78v1, 10D1_c78v2, 10D1_11B, 10D1_c85v1, 10D1_c85v2, 10D1_c85o1, 10D1_c85o2, 10D1_c87, 10D1_c90, 10D1_c91, 10D1_c92, and 10D1_c93.

[0473] In some embodiments, the antigen-binding molecule comprises:

[0474] (1) VH region containing the following CDRs:

[0475] HC-CDR1 has the amino acid sequence of SEQ ID NO:40;

[0476] HC-CDR2 has the amino acid sequence of SEQ ID NO:43;

[0477] HC-CDR3 has the amino acid sequence of SEQ ID NO:48;

[0478] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0479] VL region containing the following CDRs:

[0480] LC-CDR1 has the amino acid sequence of SEQ ID NO:66;

[0481] LC-CDR2 has the amino acid sequence of SEQ ID NO:69;

[0482] LC-CDR3 has the amino acid sequence of SEQ ID NO:74;

[0483] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0484] (2) VH region containing the following CDRs:

[0485] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0486] HC-CDR2 has the amino acid sequence of SEQ ID NO:41;

[0487] HC-CDR3 has the amino acid sequence of SEQ ID NO:44;

[0488] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0489] VL region containing the following CDRs:

[0490] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0491] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0492] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0493] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0494] (3) VH region containing the following CDRs:

[0495] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0496] HC-CDR2 has the amino acid sequence of SEQ ID NO:41;

[0497] HC-CDR3 has the amino acid sequence of SEQ ID NO:44;

[0498] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0499] VL region containing the following CDRs:

[0500] LC-CDR1 has the amino acid sequence of SEQ ID NO:64;

[0501] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0502] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0503] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0504] (4) VH region containing the following CDRs:

[0505] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0506] HC-CDR2 has the amino acid sequence of SEQ ID NO:41;

[0507] HC-CDR3 has the amino acid sequence of SEQ ID NO:44;

[0508] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0509] VL region containing the following CDRs:

[0510] LC-CDR1 has the amino acid sequence of SEQ ID NO:65;

[0511] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0512] LC-CDR3 has the amino acid sequence of SEQ ID NO:71;

[0513] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0514] (5) VH region containing the following CDRs:

[0515] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0516] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0517] HC-CDR3 has the amino acid sequence of SEQ ID NO:45;

[0518] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0519] VL region containing the following CDRs:

[0520] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0521] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0522] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0523] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0524] (6) VH region containing the following CDRs:

[0525] HC-CDR1 has the amino acid sequence of SEQ ID NO:39;

[0526] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0527] HC-CDR3 has the amino acid sequence of SEQ ID NO:45;

[0528] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0529] VL region containing the following CDRs:

[0530] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0531] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0532] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0533] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0534] (7) VH region containing the following CDRs:

[0535] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0536] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0537] HC-CDR3 has the amino acid sequence of SEQ ID NO:44;

[0538] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0539] VL region containing the following CDRs:

[0540] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0541] LC-CDR2 has the amino acid sequence of SEQ ID NO:68;

[0542] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0543] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0544] (8) VH region containing the following CDRs:

[0545] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0546] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0547] HC-CDR3 has the amino acid sequence of SEQ ID NO:46;

[0548] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0549] VL region containing the following CDRs:

[0550] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0551] LC-CDR2 has the amino acid sequence of SEQ ID NO:68;

[0552] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0553] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0554] (9) VH region containing the following CDRs:

[0555] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0556] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0557] HC-CDR3 has the amino acid sequence of SEQ ID NO:47;

[0558] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0559] VL region containing the following CDRs:

[0560] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0561] LC-CDR2 has the amino acid sequence of SEQ ID NO:68;

[0562] LC-CDR3 has the amino acid sequence of SEQ ID NO:70;

[0563] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0564] (10) VH region containing the following CDRs:

[0565] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0566] HC-CDR2 has the amino acid sequence of SEQ ID NO:42;

[0567] HC-CDR3 has the amino acid sequence of SEQ ID NO:45;

[0568] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0569] VL region containing the following CDRs:

[0570] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0571] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0572] LC-CDR3 has the amino acid sequence of SEQ ID NO:72;

[0573] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0574] (11) VH region containing the following CDRs:

[0575] HC-CDR1 has the amino acid sequence of SEQ ID NO:38;

[0576] HC-CDR2 has the amino acid sequence of SEQ ID NO:41;

[0577] HC-CDR3 has the amino acid sequence of SEQ ID NO:44;

[0578] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0579] VL region containing the following CDRs:

[0580] LC-CDR1 has the amino acid sequence of SEQ ID NO:63;

[0581] LC-CDR2 has the amino acid sequence of SEQ ID NO:67;

[0582] LC-CDR3 has the amino acid sequence of SEQ ID NO:73;

[0583] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0584] In some embodiments, the antigen-binding molecule comprises:

[0585] (12) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:21, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:49, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0586] (13) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:22, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:50, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0587] (14) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:23, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:51, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0588] (15) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:23, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:52, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0589] (16) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:23, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:53, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0590] (17) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:26, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:53, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0591] (18) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:27, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:53, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0592] (19) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:28, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:54, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0593] (20) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:29, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:54, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0594] (21) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:30, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:54, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0595] (22) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:31, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:54, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0596] (23) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:32, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:57, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0597] (24) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:33, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:58, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0598] (25) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:34, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:59, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0599] (26) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:35, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:60, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0600] (27) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:22, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:61, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0601] (28) VH region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:32, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and VL region, comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:62, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0602] In some embodiments described herein, one or more amino acids in the amino acid sequence mentioned herein (such as the amino acid sequence of an antigen-binding molecule, such as the amino acid sequence of the CDR or VH / VL region) are substituted with other amino acids. Substitution involves replacing an amino acid residue with a different “alternative” amino acid residue. The substituted amino acid residue as described in this disclosure can be a naturally occurring amino acid residue (encoded by the genetic code) and it differs from the amino acid residue at the corresponding position in the unsubstituted amino acid sequence of the equivalent. The amino acid residue is selected from: alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). In some implementations, the alternative amino acid can be a non-naturally occurring amino acid residue—that is, an amino acid residue other than those listed in the preceding sentences. Examples of non-naturally occurring amino acid residues include ornithine, ornithine, homoserine, aminoisobutyric acid, and other amino acid analogs, such as those described by Ellman et al., *Meth. Enzym*, Vol. 202, 1991, pp. 301-336.

[0603] In some embodiments, the substitution can be biochemically conserved. In some embodiments, when the amino acid to be substituted is located in one of rows 1 to 5 of the table below, the substitute amino acid is a different amino acid provided in the same row:

[0604]

[0605]

[0606] For example, in some implementations, the Met residue is replaced, and the alternative amino acids can be selected from: Ala, Val, Leu, Ile, Trp, Tyr, Phe, and ortholeucine.

[0607] In some embodiments, the substituted amino acid may have the same side chain polarity as the amino acid residue it replaces. In some embodiments, the substituted amino acid may have the same side chain charge as the amino acid residue it replaces (under pH 7.4 conditions):

[0608]

[0609]

[0610] In some embodiments, a nonpolar amino acid can be substituted with another different nonpolar amino acid. In some embodiments, a polar amino acid can be substituted with another different polar amino acid. In some embodiments, an acidic polar amino acid can be substituted with another different acidic polar amino acid. In some embodiments, a basic polar amino acid can be substituted with another different basic polar amino acid. In some embodiments, a neutral amino acid can be substituted with another different neutral amino acid. In some embodiments, a positively charged amino acid can be substituted with another different positively charged amino acid. In some embodiments, a negatively charged amino acid can be substituted with another different negatively charged amino acid.

[0611] In some implementations, the substitution may be functionally conserved. That is, in some implementations, the substitution may not affect (or will not significantly affect) one or more functional properties (such as target binding) of the antigen-binding molecule containing the substitution compared to an equivalent unsubstituted molecule.

[0612] The VH and VL regions of the antibody-antigen binding region together constitute the Fv region. In some embodiments, the antigen-binding molecule as described in this disclosure comprises or consists of an Fv region that binds to HER3. In some embodiments, the VH and VL regions of the Fv region are linked by a linker peptide region to form a single polypeptide chain, i.e., a single-chain Fv (scFv).

[0613] In some embodiments, the antigen-binding molecule as described in this disclosure comprises one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the immunoglobulin heavy chain constant sequence is or is derived from a heavy chain constant sequence of IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM.

[0614] The antigen-binding region of the antibody comprises the VL and light chain constant (CL) regions, as well as the VH and heavy chain constant 1 (CH1) regions, which together constitute the Fab region. In some embodiments, the antigen-binding molecule as described in this disclosure includes or consists of the Fab region that binds HER3.

[0615] In some embodiments, the antigen-binding molecules described herein comprise or consist of whole antibodies that bind HER3. In this document, “whole antibody” refers to an antibody having a structure substantially similar to that of immunoglobulins (Ig). Descriptions of different types of immunoglobulins and their structures are found in Schroeder and Cavacini, *J. AllergyClin. Immunol*, 2010, Vol. 125, No. 202: pp. 41-52, which are incorporated herein by reference in their entirety.

[0616] Immunoglobulins G (IgG) are glycoproteins of approximately 150 kDa, consisting of two heavy chains and two light chains. From the N-terminus to the C-terminus, the heavy chain contains the VH domain followed by a heavy chain constant region containing three constant domains (CH1, CH2, and CH3), and similarly, the light chain contains the VL domain followed by the CL domain. Based on the heavy chain, immunoglobulins can be classified as IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM. The light chains can be κ or λ type.

[0617] In some implementations, the antigen-binding molecule comprises or consists of IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM that bind to HER3.

[0618] In some embodiments, the antigen-binding molecule comprises or consists of the following:

[0619] (i) one or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence of SEQ ID NO:75; and

[0620] (ii) One or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence of SEQ ID NO:76.

[0621] The antigen-binding molecules described in this disclosure can be present in the form of compositions comprising the antigen-binding molecules. The antigen-binding molecules can be formulated into pharmaceutical compositions or agents for clinical use and may contain pharmaceutically acceptable carriers, diluents, excipients, or adjuvants. The compositions can be formulated for local, parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, oral, or transdermal administration, including injection or infusion.

[0622] Suitable formulations may contain antigen-binding molecules in a sterile or isotonic medium. Pharmaceuticals and pharmaceutical compositions may be formulated in fluid form, including gels. Fluid formulations may be formulated for administration by injection or infusion (via cannula) to a specific area of ​​the blood, tumor, or human or animal body. In some embodiments, the composition may be formulated for injection or infusion, such as injection into a blood vessel or tumor.

[0623] HER3-mediated signal transduction antagonists

[0624] The various aspects and implementations described in this disclosure involve the use of (i) HER3-mediated signal transduction antagonists and (ii) antigen-binding molecules that bind to HER3 for therapeutic / preventive intervention.

[0625] Although antigen-binding molecules that bind to HER3 as described in this disclosure can act as HER3-mediated signal transduction antagonists, it should be understood that in aspects and embodiments involving combination therapy as described in this disclosure, the combined components (i) and (ii) are preferably different from each other (i.e., they are different formulations).

[0626] Treatment with HER3-mediated signal transduction antagonists as described in this disclosure is considered, particularly in relation to therapeutic / preventive interventions for cancer treatment / prevention as described in the section above, "Cancers Characterized by the Presence of Genetic Variations Leading to Enhanced HER3-Mediated Signal Transduction." As described in such embodiments, treatment with HER3-mediated signal transduction antagonists can effectively restore signal levels to the state observed without the mutation (i.e., signal transduction levels in equivalent cells containing only the wild-type allele).

[0627] In this way, treatment with a HER3-mediated signal transduction antagonist may be effective for patients receiving the HER3-binding antigen-binding molecule therapy described herein. Specifically, administration of a HER3-mediated signal transduction antagonist preferably effectively sensitizes (i.e., susceptibles) the cancer to HER3-binding antigen-binding molecule therapy, making treatment with HER3-binding antigen-binding molecules more effective than treatment without a HER3-mediated signal transduction antagonist.

[0628] It should be understood that the specific HER3-mediated signaling antagonists used in the combination therapies described in this disclosure can be selected based on the mutational status / genotype of the cancer to be treated. For example, if the cancer contains a KRAS activating mutation, the antagonist could be a MAPK / ERK pathway signaling antagonist. Similarly, if the cancer contains a PIK3CA activating mutation or a PTEN inactivating mutation, the antagonist could be a PI3K / AKT / mTOR pathway signaling antagonist.

[0629] In some embodiments, the HER3-mediated signaling antagonist as described in this disclosure is a pan-ErbB inhibitor (such as sapitinib or Sym013). In some embodiments, the HER3-mediated signaling antagonist is an EGFR-mediated signaling inhibitor (such as cetuximab, panitumumab, gefitinib, erlotinib, lapatinib, afatinib, brigatinib, icotinib, osimertinib, zarumumab, vandetanib, nexituzumab, nimotuzumab, dacomitinib, duligotuzumab, or materutuzumab). In some embodiments, the HER3-mediated signaling antagonist is a HER2-mediated signaling inhibitor (such as trastuzumab, pertuzumab, lapatinib, neratinib, afatinib, dacomitinib, MM-111, zanotuzumab, MCLA-128, or materutuzumab). In some implementations, the HER3-mediated signal transduction antagonist is a HER3-mediated signal transduction inhibitor (such as seribamumab, rulezumab, iconumumab, KTN3379, AV-203, GSK2849330, REGN1400, MP-RM-1, EV20, pertuzumab, duligotuzumab, MM-111, zanotuzumab, istrotumumab, MCLA-128, pertrotumumab, EZN-3920, RB200, U3-1402, TX2-121-2, and miR-205). In some implementations, the HER3-mediated signal transduction antagonist is a HER4-mediated signal transduction inhibitor (such as lapatinib, ibrutinib, afatinib, dacomitinib, or neratinib).

[0630] In some embodiments, HER3-mediated signaling antagonists inhibit downstream effectors of HER3 signaling. Downstream effectors of HER3-mediated signaling include PI3K, AKT, K-Ras, β-Raf, MEK / ERK, and mTOR. In some embodiments, HER3-mediated signaling antagonists are inhibitors of the MAPK / ERK pathway. In some embodiments, HER3-mediated signaling antagonists are inhibitors of the PI3K / AKT / mTOR pathway.

[0631] In some embodiments, the HER3-mediated signal transduction antagonist is a PI3K inhibitor (such as picricidazob, bupalizib, dacrizib, SAR245409, AZD8186, ederacilib, copalizib, or duvelizib). In some embodiments, the HER3-mediated signal transduction antagonist is an AKT inhibitor (such as MK-2206, AZD5363, GSK690693, GSK2110183, ipatalizib, VQD-002, perifoxetine, or mitefoxetine). In some embodiments, the HER3-mediated signal transduction antagonist is a KRAS inhibitor (such as sotorarazib, ARS-1620, adagrazib, LY3499446, ARS-3248 / JNJ-74699157, BI2852, or RRSP chimeric toxin). In some implementations, the HER3-mediated signal transduction antagonist is a BRAF inhibitor (such as vemurafenib, dabrafenib, SB590885, XL281, RAF265, encorafenib, bevacirafenib, PLX8394, LY3009120, LXH254, GDC-0879, PLX-4720, sorafenib, or LGX818). In some implementations, the HER3-mediated signal transduction antagonist is a MEK / ERK inhibitor (such as trametinib, cobimemetinib, binimetinib, selumetinib, pimacintinib, PD-325901, CI-1040, PD035901, or TAK-733). In some implementations, the HER3-mediated signal transduction antagonist is an mTOR inhibitor (such as rapamycin, defolimus, tesirobolimus, everolimus, lidafolimus, or sapanixetil).

[0632] Treatment and preventive intervention

[0633] The various aspects and implementation schemes described in this disclosure relate to therapeutic and preventive interventions for the treatment and prevention of cancers as described herein.

[0634] This disclosure provides a HER3-binding antigen-binding molecule for use in treating or preventing the cancer described herein in a subject. It also provides the use of the HER3-binding antigen-binding molecule in the preparation of a medicament for treating or preventing the cancer described herein in a subject. A method for treating or preventing the cancer described herein in a subject is also provided, comprising administering to the subject a therapeutically effective amount or a preventatively effective amount of the HER3-binding antigen-binding molecule.

[0635] This disclosure provides a method of using a HER3-binding antigen-binding molecule for treating or preventing the cancers described herein, wherein the method further includes administering a HER3-mediated signal transduction antagonist. A method of using a HER3-mediated signal transduction antagonist for treating or preventing the cancers described herein is also provided, wherein the method further includes administering a HER3-binding antigen-binding molecule. Use of a HER3-binding antigen-binding molecule for preparing a medicament for treating or preventing the cancers described herein is also provided, wherein the treatment method further includes administering a HER3-mediated signal transduction antagonist. Use of a HER3-mediated signal transduction antagonist for preparing a medicament for treating or preventing the cancers described herein is also provided, wherein the method further includes administering a HER3-binding antigen-binding molecule. A further method of treating or preventing the cancers described herein is provided, the method comprising administering to a subject requiring treatment a therapeutically effective amount or a preventatively effective amount of (i) a HER3-binding antigen-binding molecule and (ii) a HER3-mediated signal transduction antagonist.

[0636] This disclosure also provides a method for treating or preventing the cancers described herein using a HER3-binding antigen-binding molecule and a HER3-mediated signal transduction antagonist (e.g., in the form of a pharmaceutical combination or composition comprising a HER3-binding antigen-binding molecule and a HER3-mediated signal transduction antagonist). The use of a HER3-binding antigen-binding molecule and a HER3-mediated signal transduction antagonist in the preparation of a medicament for treating or preventing the cancers described herein is also provided. A method for treating or preventing the cancers described herein is also provided, the method comprising administering to a subject requiring treatment a therapeutically effective amount or a preventatively effective amount of a HER3-binding antigen-binding molecule and a HER3-mediated signal transduction antagonist (e.g., in the form of a pharmaceutical combination or composition comprising a HER3-binding antigen-binding molecule and a HER3-mediated signal transduction antagonist).

[0637] In some aspects and implementations, HER3-binding antigen-binding molecules and HER3-mediated signal transduction antagonists can be used as combination therapy. In some implementations, HER3-binding antigen-binding molecules and HER3-mediated signal transduction antagonists can be administered simultaneously or sequentially.

[0638] Simultaneous administration refers to the administration of two or more drugs together, for example, in the form of a pharmaceutical composition containing two drugs (i.e., as a combination formulation), or immediately after one drug (e.g., within 1, 4, 6, 8, or 12 hours), or preferably via the same route, such as by application to the same artery, vein, or other blood vessel.

[0639] Sequential administration refers to administering one drug first, followed by another drug alone after a given time interval. It is not required that the drugs be administered via the same route, even if in some embodiments they are administered via the same route. The time interval can be any time interval.

[0640] In some embodiments, the therapeutic or preventative intervention as described herein comprises: (i) administering a HER3-mediated signaling antagonist (the HER3-mediated signaling antagonist described herein) to a subject suffering from cancer (cancer characterized herein by the presence of a genetic variant leading to increased HER3-mediated signaling), and (ii) administering a HER3-binding antigen-binding molecule (the HER3-binding antigen-binding molecule described herein) to the subject. In some embodiments, (i) and (ii) may be performed simultaneously. In some embodiments, (i) and (ii) may be performed sequentially (e.g., (i) after (ii), or (ii) after (i)).

[0641] The therapeutic or preventative interventions described in this disclosure can effectively reduce the development or progression of cancer, alleviate one or more symptoms of cancer, or reduce the severity of cancer symptoms. These interventions can effectively halt cancer progression, such as preventing cancer from worsening or slowing its rate of development. In some embodiments, the interventions may lead to improvements in cancer, such as reducing cancer symptoms or lowering other relevant indicators of cancer severity / activity. In some embodiments, the methods can prevent cancer from progressing to later stages (more severe stages or metastases).

[0642] In some implementations, the treatment or preventive intervention may be intended to: delay / prevent the onset / progression of cancer symptoms, reduce the severity of cancer symptoms, reduce the survival / growth / invasion / metastasis of cancer cells, reduce the number of cancer cells, and / or improve the survival rate of the subject.

[0643] In some implementations, the therapeutic or preventative interventions described in this disclosure may be associated with one or more of the following: inhibiting cancer development / progression, delaying / preventing cancer onset, reducing / delaying / preventing tumor growth, reducing / delaying / preventing tissue invasion, reducing / delaying / preventing metastasis, alleviating the severity of one or more symptoms of cancer, reducing the number of cancer cells, reducing the cancer burden, reducing tumor size / volume, and / or improving the survival rate (progression-free survival or overall survival) of subjects with the cancer.

[0644] The pharmaceutical preparations, combinations of pharmaceutical preparations, and pharmaceutical compositions described in this disclosure are preferably administered at a dose that is "therapeuticly effective" or "preventively effective," sufficient to provide the subject with a therapeutic or preventative benefit. The actual dosage, rate of administration, and duration of administration will depend on the nature and severity of the disease / symptom and the specific formulation being administered. Treatment protocols, such as dosage determination, are the responsibility of general practitioners and other physicians, and typically take into account the disease / symptom to be treated, the individual subject's condition, the site of delivery, the method of administration, and other factors known to the physician. Examples of the above techniques and protocols can be found in Remington's *The Science and Practice of Pharmacy* (edited by A. Adejare), 23rd edition (2010), Academic Press.

[0645] The routes of administration for the pharmaceutical agents described in this disclosure may include parenteral, systemic, local, intracavitary, intravascular, intravenous, intra-arterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, oral, or transdermal administration. Administration may be by injection, infusion, or swallowing.

[0646] In some aspects and embodiments, substances as described in this disclosure may be applied to target tissues or organs affected by symptoms (a condition in which tissues / organs exhibit disease / symptoms). In some aspects and embodiments, substances as described in this disclosure may be applied to the bloodstream via injection or infusion (via blood vessels) (intravenous / arterial administration), or via subcutaneous or oral routes. In some aspects and embodiments, substances as described in this disclosure may be applied to tumors.

[0647] Multiple administrations of the aforementioned pharmaceutical agents, combinations of pharmaceuticals, and pharmaceutical compositions are available. Multiple administrations can be separated by predetermined time intervals, selectable from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1, 2, 3, 4, 5, or 6 months. For example, administration can be once every 7 days, 14 days, 21 days, or 28 days (plus or minus 3, 2, or 1 day: e.g., 4, 5, 6, 8, 9, or 10 days; 11, 12, 13, 15, 16, or 17 days; 18, 19, 20, 22, 23, or 24 days; or 25, 26, 27, 29, 30, or 31 days). Treatment can be administered every 7 days, every 14 days, every 21 days, or every 28 days. Treatment intervals of approximately 7 days (plus or minus 3, 2, or 1 day), approximately 14 days (plus or minus 3, 2, or 1 day), approximately 21 days (plus or minus 3, 2, or 1 day), or approximately 28 days (plus or minus 3, 2, or 1 day) can be established between two doses.

[0648] In some embodiments, the intervention includes additional therapeutic or preventative interventions for treating / preventing cancer. In some embodiments, the therapeutic or preventative intervention is selected from chemotherapy, immunotherapy, radiotherapy, surgery, vaccination, and / or hormone therapy. In some embodiments, the therapeutic or preventative intervention includes leukapheresis. In some embodiments, the therapeutic or preventative intervention includes stem cell transplantation.

[0649] Therapeutic / preventive intervention using EGFR antagonists

[0650] The therapeutic / preventive interventions and implementation schemes described in this disclosure utilize EGFR antagonists.

[0651] This disclosure provides a method of using a HER3-binding antigen-binding molecule for treating or preventing the cancers described herein, wherein the method further includes administering an EGFR antagonist. An EGFR antagonist is also provided for using a method of treating or preventing the cancers described herein, wherein the method further includes administering a HER3-binding antigen-binding molecule.

[0652] Also provided is the use of an antigen-binding molecule binding to HER3 for the preparation of a medicament for the treatment or prevention of the cancers described herein, wherein said treatment methods further include administration of an EGFR antagonist. Also provided is the use of an EGFR antagonist for the preparation of a medicament for the treatment or prevention of the cancers described herein, wherein said treatment methods further include administration of an antigen-binding molecule binding to HER3.

[0653] A method for treating or preventing the cancer described herein is also provided, the method comprising administering to a subject requiring treatment a therapeutically effective amount or a preventatively effective amount of (i) an antigen-binding molecule that binds to HER3 and (ii) an EGFR antagonist.

[0654] This disclosure also provides (i) a HER3-binding antigen-binding molecule and (ii) an EGFR antagonist for treating or preventing the cancer described herein in a subject. Use of (i) the HER3-binding antigen-binding molecule and (ii) the EGFR antagonist in the preparation of a medicament for treating or preventing the cancer described herein in a subject is also provided. A method for treating or preventing the cancer described herein in a subject is also provided, comprising administering to the subject a therapeutically effective amount or a preventively effective amount of (i) the HER3-binding antigen-binding molecule and (ii) the EGFR antagonist.

[0655] In the embodiments described above, (i) and (ii) may be used as a combination therapy. In some embodiments, (i) and (ii) may be provided simultaneously or sequentially.

[0656] In some implementations, an EGFR antagonist is an inhibitor of one or more EGFR functions / activities, or an inhibitor of EGFR expression (i.e., gene and / or protein expression). EGFR antagonists include inhibitors of EGFR-mediated signaling.

[0657] In some implementations, EGFR antagonists bind to EGFR. EGFR-binding EGFR antagonists can inhibit one or more functions of EGFR. EGFR-binding EGFR antagonists include EGFR-binding antibodies and their antigen-binding fragments and derivatives, EGFR-binding peptides / peptides, EGFR-binding nucleic acid aptamers, and EGFR-binding small molecule inhibitors.

[0658] In some implementations, EGFR antagonists are agents that inhibit EGFR expression. These EGFR antagonists include small molecule inhibitors of EGFR expression and nucleic acids (antisense oligonucleotides) capable of blocking or reducing EGFR expression.

[0659] In some embodiments, the EGFR antagonist as described in this disclosure is a small molecule EGFR inhibitor. In some embodiments, the EGFR antagonist is a pan-ErbB inhibitor (such as sapitinib or Sym013). In some embodiments, the EGFR antagonist is selected from gefitinib, erlotinib, lapatinib, afatinib, brigatinib, icotinib, osimertinib, vandetanib, and dacomitinib.

[0660] In some embodiments, the EGFR antagonist is an antigen-binding molecule that binds to EGFR, such as an EGFR-binding antibody or its antigen-binding fragment or derivative. In some embodiments, the EGFR antagonist is selected from cetuximab, panitumumab, zarumumab, duligotuzumab, and mateuzumab.

[0661] In some embodiments, the EGFR-binding antigen-binding molecule includes a CDR region capable of binding EGFR. In some embodiments, the antigen-binding molecule includes an FR region capable of binding EGFR. In some embodiments, the antigen-binding molecule includes both a CDR region and an FR region capable of binding EGFR. That is, in some embodiments, the antigen-binding molecule includes a VH region and a VL region capable of binding EGFR.

[0662] In some embodiments, the antigen-binding molecule capable of binding EGFR, as described in this disclosure, may be selected from: any embodiment of the antigen-binding molecule described in US6217866B1 (which is incorporated herein by reference in its entirety), cetuximab (see US6217866B1 and Wong et al., *Clin Ther.*, 2005, Vol. 27, No. 6, pp. 684-694; Pharmacy Database No. DB00002), panitumumab (see Foon et al., *International Journal of Radiation Oncology Biophysics*, 2004, Vol. 58, No. 3, pp. 984-990; Pharmacy Database No. DB01269), zarumumab (see Bastholt et al., *Radiother Oncol.*, 2007, Vol. 85, No. 1, pp. 24-28; Pharmacy Database No. DB12202), and nexituzumab (see Kuenen et al., *Clinical Cancer Research*). Res., 2010, Vol. 16, No. 6, pp. 1915-1923; Drug Library Registry No. DB09559), nimotuzumab (see Ramakrishnan et al., Monoclonal Antibodies (mAbs), 2009, Vol. 1, No. 1, pp. 41-48; Drug Library Registry No. DB06192), duligotuzumab (see Fayette et al., Frontiers in Oncology, 2016, Vol. 6, p. 232; Drug Library Registry No. DB12142), and mateuzumab (Drug Library Registry No. DB05101). In some embodiments, the antigen-binding molecule is cetuximab.

[0663] In some embodiments, the antigen-binding molecule is capable of binding to the same region or overlapping region of EGFR to a region of EGFR that binds to an antibody containing the VH and VL sequences of cetuximab.

[0664] In some embodiments, the antigen-binding molecule comprises the CDR region of cetuximab or comprises its VH and VL regions.

[0665] In some implementations, the antigen-binding molecule that binds to EGFR includes:

[0666] VH region containing the following CDRs:

[0667] HC-CDR1 has the amino acid sequence of SEQ ID NO:78;

[0668] HC-CDR2 has the amino acid sequence of SEQ ID NO:79;

[0669] HC-CDR3 has the amino acid sequence of SEQ ID NO:80;

[0670] Or variants thereof, wherein one, two, or three amino acids of one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with other amino acids; and

[0671] VL region containing the following CDRs:

[0672] LC-CDR1 has the amino acid sequence of SEQ ID NO:82;

[0673] LC-CDR2 has the amino acid sequence of SEQ ID NO:83;

[0674] LC-CDR3 has the amino acid sequence of SEQ ID NO:84;

[0675] Or its variants, wherein one, two, or three amino acids of one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced by other amino acids.

[0676] In some implementations, the antigen-binding molecule that binds to EGFR includes:

[0677] The VH region comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:77, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; and

[0678] VL region comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:81, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity.

[0679] In some implementations, the antigen-binding molecule that binds to EGFR comprises or consists of the following:

[0680] (i) one or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence of SEQ ID NO:85; and

[0681] (ii) One or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity with the amino acid sequence of SEQ ID NO:86.

[0682] In some embodiments, the antigen-binding molecule that binds to EGFR is cetuximab.

[0683] Diagnostic and prognostic methods and subject screening

[0684] This disclosure also provides diagnostic, prognostic, and predictive methods related to the cancers described herein.

[0685] The method can be performed in vitro on samples obtained from a subject, or after processing the samples obtained from a subject. Since the in vitro method does not require the presence of a subject after sample collection, it does not necessarily have to be performed on a human or animal. However, in some embodiments, the method can also be performed in vivo.

[0686] Samples may be taken from any tissue or bodily fluid. Samples may include or be derived from: a volume of blood; a volume of serum derived from the subject's blood, which may include the fluid portion of blood obtained after removing fibrin clots and blood cells; tissue samples or biopsy tissue; pleural fluid; cerebrospinal fluid (CSF); or cells isolated from the subject. In some embodiments, samples may be obtained from or derived from one or more tissues affected by a disease / condition (where one or more tissues exhibit disease symptoms or are related to the pathogenesis of the disease / condition). In some embodiments, samples may be obtained from or derived from cancer, tumors, or their cells.

[0687] The methods described herein can be used for the purpose of diagnosing cancer (the cancers described herein). The methods can be used for the purpose of prognostic / predictive assessment of a subject's likely response to the therapeutic / preventive interventions described herein. The methods can be used to predict the likely response to a given therapeutic / preventive intervention, such as in terms of efficacy, and thus may help support clinical decision-making. The methods can be used to identify / screen subjects suitable for the therapeutic / preventive interventions described herein.

[0688] In some aspects and implementations, the method includes analyzing a subject's cancer to determine whether the cancer is the cancer described herein, such as: cancer as described in the section "Cancer characterized by the absence of genetic variants leading to enhanced HER3-mediated signaling", or cancer as described in the section "Cancer characterized by the presence of genetic variants leading to enhanced HER3-mediated signaling", or cancer as described in the section "Cancer characterized by the presence of genetic variants leading to increased expression of genes on the long arm of chromosome 3 or enh...

Claims

1. An antigen binding molecule that binds HER3 for use in a method of treating or preventing a HER3 -associated cancer in a subject, wherein the HER3 -associated cancer does not comprise a genetic variation that results in increased expression of MET or enhanced activity of a gene product.

2. Use of an antigen binding molecule that binds HER3 for the manufacture of a medicament for treating or preventing a HER3 -associated cancer in a subject, wherein the HER3 -associated cancer does not comprise a genetic variation that results in increased expression of MET or enhanced activity of a gene product.

3. A method of treating or preventing a HER3 -associated cancer in a subject comprising administering to the subject a therapeutically or prophylactically effective amount of an antigen binding molecule that binds HER3, wherein the HER3 -associated cancer does not comprise a genetic variation that results in increased expression of MET or enhanced activity of a gene product.

4. The use of an antigen binding molecule as claimed in claim 1, the use as claimed in claim 2, or the method as claimed in claim 3, wherein the HER3 -associated cancer: (i) does not comprise a genetic variation that results in increased expression of KRAS or enhanced activity of a gene product; or (ii) does not comprise a genetic variation that results in increased expression of PIK3CA or enhanced activity of a gene product; or (iii) does not comprise a genetic variation that results in increased expression of BRAF or enhanced activity of a gene product; or (iv) does not comprise a genetic variation that results in decreased expression of PTEN or reduced activity of a gene product.

5. The antigen binding molecule as claimed in claim 1 or claim 4, the use as claimed in claim 2 or claim 4, or the method as claimed in claim 3 or claim 4, wherein the HER3 -associated cancer: (i) does not comprise a genetic variation that results in increased expression of KRAS or enhanced activity of a gene product; and (ii) does not comprise a genetic variation that results in increased expression of PIK3CA or enhanced activity of a gene product; and (iii) does not comprise a genetic variation that results in increased expression of BRAF or enhanced activity of a gene product; and (iv) does not comprise a genetic variation that results in decreased expression of PTEN or reduced activity of a gene product.

6. The antigen binding molecule as claimed in any one of claims 1, 4 or 5, the use as claimed in any one of claims 2, 4 or 5, or the method as claimed in any one of claims 3 to 5, wherein the HER3 -associated cancer: (i) does not comprise a MET gene amplification; and (ii) does not comprise a KRAS activating mutation; and (iii) does not comprise a PIK3CA activating mutation; and (iv) does not comprise a BRAF activating mutation; and (v) does not comprise a PTEN gene deletion.

7. The antigen binding molecule as claimed in any one of claims 1 or 4 to 6, the use as claimed in any one of claims 2 or 4 to 6, or the method as claimed in any one of claims 3 to 6, wherein the HER3 -associated cancer comprises an amplification of one or more genes located on the long arm of chromosome 3, optionally the one or more genes located on the long arm of chromosome 3 are located within chromosome 3q26-3q28, optionally the one or more genes are selected from: TP63, SOX2 and PIK3CA.

8. The antigen binding molecule of any one of claims 1 or 4 to 7, the use of any one of claims 2 or 4 to 7, or the method of any one of claims 3 to 7, wherein the HER3-associated cancer comprises amplification of one or more genes located on the short arm of chromosome 7, optionally the one or more genes are located within chromosome 7pl l, optionally the gene is EGFR.

9. The antigen binding molecule of any one of claims 1 or 4 to 7, the use of any one of claims 2 or 4 to 7, or the method of any one of claims 3 to 7, wherein the HER3-associated cancer comprises deletion of one or more genes located on the short arm of chromosome 3, optionally the one or more genes are located within chromosome 3p21, optionally the gene is TUSC2.

10. The antigen binding molecule of any one of claims 1 or 4 to 9, the use of any one of claims 2 or 4 to 9, or the method of any one of claims 3 to 9, wherein the HER3-associated cancer comprises a genetic variation that results in increased expression of a HER3 ligand.

11. The antigen binding molecule of any one of claims 1 or 4 to 10, the use of any one of claims 2 or 4 to 10, or the method of any one of claims 3 to 10, wherein the HER3-associated cancer comprises a NRG gene fusion, a NRG1 gene fusion, or a NRG2 gene fusion.

12. The antigen binding molecule of any one of claims 1 or 4 to 11, the use of any one of claims 2 or 4 to 11, or the method of any one of claims 3 to 11, wherein the HER3-associated cancer comprises a NRG gene fusion selected from the group consisting of CLU-NRG1, CD74-NRG1, DOC4-NRG1, SLC3A2-NRG1, RBPMS-NRG1, WRN-NRG1, SDC4-NRG1, RAB2IL1-NRG1, VAMP2-NRG1, KIF13B-NRG1, THAP7-NRG1, SMAD4-NRG1, MDK-NRG1, TNC-NRG1, DIP2B-NRG1, MRPL13-NRG1, PARP8-NRG1, ROCK1-NRG1, DPYSL2-NRG1, ATP1B1-NRG1, CDH6-NRG1, APP-NRG1, AKAP13-NRG1, THBS1-NRG1, FOXA1-NRG1, PDE7A-NRG1, RAB3IL1-NRG1, CDK1-NRG1, BMPRIB-NRG1, TNFRSF10B-NRG1, MCPH1-NRG1, and SLC12A2-NRG2.

13. An antigen binding molecule that binds HER3 for use in a method of treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises amplification of one or more genes located on the long arm of chromosome 3, optionally the one or more genes located on the long arm of chromosome 3 are located within chromosome 3q26-3q28, optionally the one or more genes are selected from the group consisting of: TP63, SOX2, and PIK3CA.

14. Use of an antigen binding molecule that binds HER3 in the manufacture of a medicament for treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises amplification of one or more genes located on the long arm of chromosome 3, optionally the one or more genes located on the long arm of chromosome 3 are located within chromosome 3q26-3q28, optionally the one or more genes are selected from the group consisting of: TP63, SOX2, and PIK3CA.

15. A method of treating or preventing a HER3-associated cancer in a subject, comprising administering to the subject a therapeutically effective amount or a prophylactically effective amount of an antigen binding molecule that binds HER3, wherein the HER3-associated cancer comprises amplification of one or more genes located on the long arm of chromosome 3, optionally the one or more genes located on the long arm of chromosome 3 are located within chromosome 3q26-3q28, optionally the one or more genes are selected from the group consisting of: TP63, SOX2, and PIK3CA.

16. The antigen binding molecule of claim 13 or claim 16, the use of claim 14 or claim 16, or the method of claim 15 or claim 16, wherein the HER3-associated cancer comprises TP63 gene amplification, and comprises SOX2 gene amplification, and comprises PIK3CA gene amplification.

17. An antigen binding molecule that binds HER3 for use in a method of treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises amplification of one or more genes located on the short arm of chromosome 7, optionally the one or more genes are located within chromosome 7pl l, optionally the gene is EGFR.

18. Use of an antigen binding molecule that binds HER3 in the manufacture of a medicament for treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises amplification of one or more genes located on the short arm of chromosome 7, optionally the one or more genes are located within chromosome 7pl l, optionally the gene is EGFR.

19. A method of treating or preventing a HER3-associated cancer in a subject, comprising administering to the subject a therapeutically effective amount or a prophylactically effective amount of an antigen binding molecule that binds HER3, wherein the HER3-associated cancer comprises amplification of one or more genes located on the short arm of chromosome 7, optionally the one or more genes are located within chromosome 7pl l, optionally the gene is EGFR.

20. An antigen binding molecule that binds HER3 for use in a method of treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises a deletion of one or more genes located on the short arm of chromosome 3, optionally the one or more genes are located within chromosome 3p21, optionally the gene is TUSC2.

21. Use of an antigen binding molecule that binds HER3 in the manufacture of a medicament for treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises a deletion of one or more genes located on the short arm of chromosome 3, optionally the one or more genes are located within chromosome 3p21, optionally the gene is TUSC2.

22. A method of treating or preventing a HER3-associated cancer in a subject, comprising administering to the subject a therapeutically effective amount or a prophylactically effective amount of an antigen binding molecule that binds HER3, wherein the HER3-associated cancer comprises a deletion of one or more genes located on the short arm of chromosome 3, optionally the one or more genes are located within chromosome 3p21, optionally the gene is TUSC2.

23. An antigen binding molecule that binds HER3 for use in a method of treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises a genetic variation that results in increased expression of MET or enhanced activity of a gene product, and the method further comprises administering an antagonist of HER3-mediated signaling.

24. Use of an antigen binding molecule that binds HER3 in the manufacture of a medicament for treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer comprises a genetic variation that results in increased expression of MET or enhanced activity of a gene product, and the method of treatment further comprises administering an antagonist of HER3-mediated signaling.

25. A method of treating or preventing a HER3-associated cancer in a subject, comprising administering to the subject a therapeutically effective amount or a prophylactically effective amount of an antigen binding molecule that binds HER3, wherein the HER3-associated cancer comprises a genetic variation that results in increased expression of MET or enhanced activity of a gene product, and the method further comprises administering an antagonist of HER3-mediated signaling.

26. The antigen binding molecule of claim 23, the use of claim 24, or the method of claim 25, wherein the HER3-associated cancer: (i) comprises a genetic variation that results in increased expression of KRAS or enhanced activity of a gene product; or (ii) comprises a genetic variation that results in increased expression of PIK3CA or enhanced activity of a gene product; or (iii) comprises a genetic variation that results in increased expression of BRAF or enhanced activity of a gene product; or (iv) comprises a genetic variation that results in decreased expression of PTEN or reduced activity of a gene product.

27. An antigen binding molecule as claimed in claim 23 or claim 26, use as claimed in claim 24 or claim 26, or a method as claimed in claim 25 or claim 26, wherein the HER3- associated cancer: (i) comprises a genetic variation that results in increased expression or enhanced activity of the gene product of KRAS; and (ii) comprises a genetic variation that results in increased expression or enhanced activity of the gene product of PIK3CA; and (iii) comprises a genetic variation that results in increased expression or enhanced activity of the gene product of BRAF; and (iv) comprises a genetic variation that results in decreased expression or reduced activity of the gene product of PTEN.

28. A method of selecting a subject for treatment with an antigen binding molecule that binds HER3, comprising: (a) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in increased expression or enhanced activity of the gene product of MET; and (b) selecting the subject for treatment with an antigen binding molecule that binds HER3 if step (a) determines that the subject's cancer does not comprise such a genetic variation.

29. A method as claimed in claim 28, wherein the method comprises: (i) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in increased expression or enhanced activity of the gene product of KRAS; or (a) (ii) analysing the subject's cancer to determine whether the cancer comprises a PIK3CA activating mutation; or (iii) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in increased expression or enhanced activity of the gene product of BRAF; or (iv) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in decreased expression or reduced activity of the gene product of PTEN; and (b) selecting the subject for treatment with an antigen binding molecule that binds HER3 if step (a) determines that the subject's cancer does not comprise such a genetic variation / mutation.

30. A method as claimed in claim 28 or claim 29, wherein the method comprises: (i) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in increased expression or enhanced activity of the gene product of KRAS; and (a) (ii) analysing the subject's cancer to determine whether the cancer comprises a PIK3CA activating mutation; and (iii) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in increased expression or enhanced activity of the gene product of BRAF; and (iv) analysing the subject's cancer to determine whether the cancer comprises a genetic variation that results in decreased expression or reduced activity of the gene product of PTEN; and (b) selecting the subject for treatment with an antigen binding molecule that binds HER3 if step (a) determines that the subject's cancer does not comprise such a genetic variation / mutation.

31. A method as claimed in any one of claims 28 to 30, wherein the method further comprises: (c) administering an antigen binding molecule that binds HER3 to the subject selected in step (b) for treatment. ​ ​ ​ 32. A method of selecting a subject for treatment with (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3, comprising: (a) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in increased expression of MET or enhanced activity of a gene product; and (b) selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3 if step (a) determines that the cancer of the subject comprises such a genetic variation.

33. The method of claim 32, wherein the method comprises: (a) (i) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in increased expression of KRAS or enhanced activity of a gene product; or (ii) analyzing a cancer of the subject to determine whether the cancer comprises a PIK3CA activating mutation; or (iii) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in increased expression of BRAF or enhanced activity of a gene product; or (iv) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in decreased expression of PTEN or reduced activity of a gene product; and (b) selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3 if step (a) determines that the cancer of the subject comprises such a genetic variation / mutation.

34. The method of claim 32 or claim 33, wherein the method comprises: (a) (i) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in increased expression of KRAS or enhanced activity of a gene product; and (ii) analyzing a cancer of the subject to determine whether the cancer comprises a PIK3CA activating mutation; and (iii) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in increased expression of BRAF or enhanced activity of a gene product; and (iv) analyzing a cancer of the subject to determine whether the cancer comprises a genetic variation that results in decreased expression of PTEN or reduced activity of a gene product; and (b) selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3 if step (a) determines that the cancer of the subject comprises such a genetic variation / mutation.

35. The method of any one of claims 32 to 34, wherein the method further comprises: (c) administering (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3 to the subject selected in step (b) to receive treatment.

36. The antigen binding molecule of any one of claims 1, 4 to 13, 16, 17, 20, 23, 26, or 27, the use of any one of claims 2, 4 to 12, 14, 16, 18, 21, 24, 26, or 27, or the method of any one of claims 3 to 12, 15, 19, 22, 25, or 28 to 35, wherein the HER3 -associated cancer is selected from the group consisting of a solid tumor, a breast cancer, a breast carcinoma, a ductal carcinoma, a gastric cancer, a gastric carcinoma, a gastric adenocarcinoma, a colorectal cancer, a colorectal carcinoma, a colorectal adenocarcinoma, a head and neck cancer, a head and neck squamous cell carcinoma, a lung cancer, a non-small cell lung cancer, a lung adenocarcinoma, a lung squamous cell carcinoma, an ovarian cancer, an ovarian carcinoma, an ovarian serous adenocarcinoma, a renal cancer, a renal cell carcinoma, a renal clear cell carcinoma, a renal cell adenocarcinoma, a renal papillary cell carcinoma, a pancreatic cancer, a pancreatic adenocarcinoma, a pancreatic ductal adenocarcinoma, a cervical cancer, a cervical squamous cell carcinoma, a skin cancer, a melanoma, an esophageal cancer, an esophageal adenocarcinoma, a liver cancer, a hepatocellular carcinoma, a cholangiocarcinoma, a uterine cancer, a uterine corpus endometrial carcinoma, a thyroid cancer, a thyroid carcinoma, a pheochromocytoma, a paraganglioma, a bladder cancer, a bladder urothelial carcinoma, a prostate cancer, a prostate adenocarcinoma, a sarcoma, and a thymoma.

37. The antigen binding molecule of any one of claims 1, 4 to 13, 16, 17, 20, 23, 26, 27, or 36, the use of any one of claims 2, 4 to 12, 14, 16, 18, 21, 24, 26, 27, or 36, or the method of any one of claims 3 to 12, 15, 19, or 22 to 36, wherein the antigen binding molecule that binds HER3 is selected from the group consisting of 10D1F, seribantumab, elgemtumab, patritumab, GSK2849330, lumretuzumab, CDX-3379, AV-203, barecetamab, TK-A3, TK-A4, MP-EV20, 1A5-3D4, 9F7-F11, 16D3-C1, NG33, A5, F4, huHER3-8, REGN1400, and zenocutuzumab.

38. Use of an antigen binding molecule of any one of claims 1, 4-13, 16, 17, 20, 23, 26, 27, 36, or 37, use of any one of claims 2, 4-12, 14, 16, 18, 21, 24, 26, 27, 36, or 37, or a method of any one of claims 3-12, 15, 19, or 22-37, wherein the antigen binding molecule that binds HER3 comprises: (i) a heavy chain variable (VH) region comprising the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 40; HC-CDR2 having the amino acid sequence of SEQ ID NO: 43; HC-CDR3 having the amino acid sequence of SEQ ID NO: 48; and (ii) a light chain variable (VL) region comprising the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 66; LC-CDR2 having the amino acid sequence of SEQ ID NO: 69; LC-CDR3 having the amino acid sequence of SEQ ID NO:

74.

39. An antigen binding molecule of any one of claims 1, 4-13, 16, 17, 20, 23, 26, 27, or 36-38, use of any one of claims 2, 4-12, 14, 16, 18, 21, 24, 26, 27, or 36-38, or a method of any one of claims 3-12, 15, 19, or 22-38, wherein the antigen binding molecule that binds HER3 comprises: (i) a VH region comprising the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38; HC-CDR2 having the amino acid sequence of SEQ ID NO: 42; HC-CDR3 having the amino acid sequence of SEQ ID NO: 45; and (ii) a VL region comprising the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63; LC-CDR2 having the amino acid sequence of SEQ ID NO: 67; LC-CDR3 having the amino acid sequence of SEQ ID NO:

70.

40. An antigen binding molecule of any one of claims 1, 4-13, 16, 17, 20, 23, 26, 27, or 36-39, use of any one of claims 2, 4-12, 14, 16, 18, 21, 24, 26, 27, or 36-39, or a method of any one of claims 3-12, 15, 19, or 22-39, wherein the antigen binding molecule that binds HER3 comprises: a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 33; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

58.

41. The antigen binding molecule of any one of claims 1, 4 to 13, 16, 17, 20, 23, 26, 27, or 36 to 40, the use of any one of claims 2, 4 to 12, 14, 16, 18, 21, 24, 26, 27, or 36 to 40, or the method of any one of claims 3 to 12, 15, 19, or 22 to 40, wherein the antigen binding molecule that binds HER3 comprises: a polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 75; and a polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

76.

42. The antigen binding molecule of any one of claims 1, 4 to 13, 16, 17, 20, 23, 26, 27, or 36 to 41, the use of any one of claims 2, 4 to 12, 14, 16, 18, 21, 24, 26, 27, or 36 to 41, or the method of any one of claims 3 to 12, 15, 19, or 22 to 41, wherein the method of treating or preventing a HER3-associated cancer further comprises administering to the subject an EGFR antagonist.

43. The antigen binding molecule of claim 42, the use of claim 42, or the method of claim 42, wherein the EGFR antagonist is an antigen binding molecule that binds EGFR.

44. The antigen binding molecule of claim 43, the use of claim 43, or the method of claim 43, wherein the antigen binding molecule that binds EGFR comprises: (i) a VH region comprising the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 78; HC-CDR2 having the amino acid sequence of SEQ ID NO: 79; HC-CDR3 having the amino acid sequence of SEQ ID NO: 80; and (ii) a VL region comprising the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 82; LC-CDR2 having the amino acid sequence of SEQ ID NO: 83; LC-CDR3 having the amino acid sequence of SEQ ID NO:

84.

45. The antigen binding molecule of claim 43 or claim 44, the use of claim 43 or claim 44, or the method of claim 43 or claim 44, wherein the antigen binding molecule that binds EGFR comprises: a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 77; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

81.

46. ​​The antigen-binding molecule of any one of claims 43 to 45, the use of any one of claims 43 to 45, or the method of any one of claims 43 to 45, wherein the EGFR-binding antigen-binding molecule comprises: A polypeptide comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:85; and A polypeptide comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:86.

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