T cells can be redirected to target tumor-associated antigen (TAA) by genetic modification to express a chimeric antigen receptor (CAR), which fuses the specificity derived from an antibody to T-cell activation domains to result in lysis of TAA-expressing cells. Due to the potential for on-target, off-tissue toxicity, CAR+ T-cell therapy is currently limited to unique or lineage-restricted TAAs. Glioblastoma, a grade IV brain malignancy, overexpresses epidermal growth factor receptor (EGFR) in 40-50% of patients. EGFR also has widespread normal tissue expression. To target EGFR on glioblastoma while reducing the potential for normal tissue toxicity, EGFR-specific CAR generated from cetuximab, Cetux-CAR, was transiently expressed in T cells by RNA-modification. RNA-modified CAR+ T cells demonstrated similar cytotoxicity against EGFR+ cells, including normal renal cells, as DNA-modified CAR+ T cells. However, RNA-modified T cells lost CAR expression over time, concomitant with loss of functional specificity to EGFR. Transient expression of CAR limits potential for off-tissue toxicity at the expense of anti-tumor activity, and does not protect normal tissue from immediate toxicity. Recognizing that EGFR is overexpressed at a higher density on glioblastoma relative to normal tissue, we generated an EGFR-specific CAR from nimotuzumab, an EGFR-specific antibody with reduced binding to low density EGFR. While Cetux-CAR+ T cells produced cytokine and mediated lysis independent of EGFR density, function of Nimo-CAR+ T cells directly correlated with the EGFR density of targets, with reduced activity in response to low density EGFR, but equivalent activity in response to high density EGFR relative to Cetux-CAR+ T cells. Cetux-CAR+ T cells and Nimo-CAR+ T cells demonstrated equivalent control of intracranial glioma xenograft with intermediate EGFR density, but only Cetux-CAR+ T cells controlled xenografts with low EGFR density. In sum, transient expression of CAR has the potential to reduce long-term toxicity to normal tissue, but at the expense of anti-tumor activity. Rational design of CAR based on an antibody with reduced binding to low density EGFR generated EGFR-specific CAR able to tune T-cell function to antigen density resulting in discrimination of high EGFR density on malignant cells from low EGFR density on normal tissue.
","abstract_html":"<p>T cells can be redirected to target tumor-associated antigen (TAA) by genetic modification to express a chimeric antigen receptor (CAR), which fuses the specificity derived from an antibody to T-cell activation domains to result in lysis of TAA-expressing cells. Due to the potential for on-target, off-tissue toxicity, CAR<sup>+</sup> T-cell therapy is currently limited to unique or lineage-restricted TAAs. Glioblastoma, a grade IV brain malignancy, overexpresses epidermal growth factor receptor (EGFR) in 40-50% of patients. EGFR also has widespread normal tissue expression. To target EGFR on glioblastoma while reducing the potential for normal tissue toxicity, EGFR-specific CAR generated from cetuximab, Cetux-CAR, was transiently expressed in T cells by RNA-modification. RNA-modified CAR<sup>+</sup> T cells demonstrated similar cytotoxicity against EGFR<sup>+</sup> cells, including normal renal cells, as DNA-modified CAR<sup>+</sup> T cells. However, RNA-modified T cells lost CAR expression over time, concomitant with loss of functional specificity to EGFR. Transient expression of CAR limits potential for off-tissue toxicity at the expense of anti-tumor activity, and does not protect normal tissue from immediate toxicity. Recognizing that EGFR is overexpressed at a higher density on glioblastoma relative to normal tissue, we generated an EGFR-specific CAR from nimotuzumab, an EGFR-specific antibody with reduced binding to low density EGFR. While Cetux-CAR<sup>+</sup> T cells produced cytokine and mediated lysis independent of EGFR density, function of Nimo-CAR<sup>+</sup> T cells directly correlated with the EGFR density of targets, with reduced activity in response to low density EGFR, but equivalent activity in response to high density EGFR relative to Cetux-CAR<sup>+</sup> T cells. Cetux-CAR<sup>+</sup> T cells and Nimo-CAR<sup>+</sup> T cells demonstrated equivalent control of intracranial glioma xenograft with intermediate EGFR density, but only Cetux-CAR<sup>+</sup> T cells controlled xenografts with low EGFR density. In sum, transient expression of CAR has the potential to reduce long-term toxicity to normal tissue, but at the expense of anti-tumor activity. Rational design of CAR based on an antibody with reduced binding to low density EGFR generated EGFR-specific CAR able to tune T-cell function to antigen density resulting in discrimination of high EGFR density on malignant cells from low EGFR density on normal tissue.</p>","abstract_has_math":false,"creators":["Caruso, Hillary G"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation (PhD)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Laurence Cooper, M.D., Ph.D.","Oliver Bogler, Ph.D.","Bradley McIntyre, Ph.D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-05-01T07:00:00Z","date_published":"2014-05-01T07:00:00Z","updated_at":"2026-07-24T05:49:23Z","subjects":["genetically modified T cells","cancer immunotherapy","affinity","EGFR","glioma","Cancer Biology","Immunity","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/457","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Laurence Cooper, M.D., Ph.D.","Oliver Bogler, Ph.D.","Bradley McIntyre, Ph.D."]},{"key":"dc:creator","label":"Author","values":["Caruso, Hillary G"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2014-05-03T07:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation (PhD)"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["genetically modified T cells","cancer immunotherapy","affinity","EGFR","glioma","Cancer Biology","Immunity","Medicine and Health Sciences"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/457"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["T cells can be redirected to target tumor-associated antigen (TAA) by genetic modification to express a chimeric antigen receptor (CAR), which fuses the specificity derived from an antibody to T-cell activation domains to result in lysis of TAA-expressing cells. Due to the potential for on-target, off-tissue toxicity, CAR+ T-cell therapy is currently limited to unique or lineage-restricted TAAs. Glioblastoma, a grade IV brain malignancy, overexpresses epidermal growth factor receptor (EGFR) in 40-50% of patients. EGFR also has widespread normal tissue expression. To target EGFR on glioblastoma while reducing the potential for normal tissue toxicity, EGFR-specific CAR generated from cetuximab, Cetux-CAR, was transiently expressed in T cells by RNA-modification. RNA-modified CAR+ T cells demonstrated similar cytotoxicity against EGFR+ cells, including normal renal cells, as DNA-modified CAR+ T cells. However, RNA-modified T cells lost CAR expression over time, concomitant with loss of functional specificity to EGFR. Transient expression of CAR limits potential for off-tissue toxicity at the expense of anti-tumor activity, and does not protect normal tissue from immediate toxicity. Recognizing that EGFR is overexpressed at a higher density on glioblastoma relative to normal tissue, we generated an EGFR-specific CAR from nimotuzumab, an EGFR-specific antibody with reduced binding to low density EGFR. While Cetux-CAR+ T cells produced cytokine and mediated lysis independent of EGFR density, function of Nimo-CAR+ T cells directly correlated with the EGFR density of targets, with reduced activity in response to low density EGFR, but equivalent activity in response to high density EGFR relative to Cetux-CAR+ T cells. Cetux-CAR+ T cells and Nimo-CAR+ T cells demonstrated equivalent control of intracranial glioma xenograft with intermediate EGFR density, but only Cetux-CAR+ T cells controlled xenografts with low EGFR density. In sum, transient expression of CAR has the potential to reduce long-term toxicity to normal tissue, but at the expense of anti-tumor activity. Rational design of CAR based on an antibody with reduced binding to low density EGFR generated EGFR-specific CAR able to tune T-cell function to antigen density resulting in discrimination of high EGFR density on malignant cells from low EGFR density on normal tissue.
"]},{"key":"dc:title","label":"Title","values":["Car-Modified T Cells Capable of Distinguishing Normal Cells From Malignant Cells"]}]}],"canonical_facts":{"dc:contributor":["Laurence Cooper, M.D., Ph.D.","Oliver Bogler, Ph.D.","Bradley McIntyre, Ph.D."],"dc:creator":["Caruso, Hillary G"],"dc:date.available":["2014-05-03T07:00:00Z"],"dc:description.abstract":["T cells can be redirected to target tumor-associated antigen (TAA) by genetic modification to express a chimeric antigen receptor (CAR), which fuses the specificity derived from an antibody to T-cell activation domains to result in lysis of TAA-expressing cells. Due to the potential for on-target, off-tissue toxicity, CAR+ T-cell therapy is currently limited to unique or lineage-restricted TAAs. Glioblastoma, a grade IV brain malignancy, overexpresses epidermal growth factor receptor (EGFR) in 40-50% of patients. EGFR also has widespread normal tissue expression. To target EGFR on glioblastoma while reducing the potential for normal tissue toxicity, EGFR-specific CAR generated from cetuximab, Cetux-CAR, was transiently expressed in T cells by RNA-modification. RNA-modified CAR+ T cells demonstrated similar cytotoxicity against EGFR+ cells, including normal renal cells, as DNA-modified CAR+ T cells. However, RNA-modified T cells lost CAR expression over time, concomitant with loss of functional specificity to EGFR. Transient expression of CAR limits potential for off-tissue toxicity at the expense of anti-tumor activity, and does not protect normal tissue from immediate toxicity. Recognizing that EGFR is overexpressed at a higher density on glioblastoma relative to normal tissue, we generated an EGFR-specific CAR from nimotuzumab, an EGFR-specific antibody with reduced binding to low density EGFR. While Cetux-CAR+ T cells produced cytokine and mediated lysis independent of EGFR density, function of Nimo-CAR+ T cells directly correlated with the EGFR density of targets, with reduced activity in response to low density EGFR, but equivalent activity in response to high density EGFR relative to Cetux-CAR+ T cells. Cetux-CAR+ T cells and Nimo-CAR+ T cells demonstrated equivalent control of intracranial glioma xenograft with intermediate EGFR density, but only Cetux-CAR+ T cells controlled xenografts with low EGFR density. In sum, transient expression of CAR has the potential to reduce long-term toxicity to normal tissue, but at the expense of anti-tumor activity. Rational design of CAR based on an antibody with reduced binding to low density EGFR generated EGFR-specific CAR able to tune T-cell function to antigen density resulting in discrimination of high EGFR density on malignant cells from low EGFR density on normal tissue.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/457"],"dc:subject":["genetically modified T cells","cancer immunotherapy","affinity","EGFR","glioma","Cancer Biology","Immunity","Medicine and Health Sciences"],"dc:title":["Car-Modified T Cells Capable of Distinguishing Normal Cells From Malignant Cells"],"thesis:degree_level":["Dissertation (PhD)"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T05:49:23Z"}