{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T12:03:47Z","timestamp":1791201827206,"version":"4.2.0"},"posted":{"date-parts":[[2025,11,17]]},"group-title":"Cancer Biology","reference-count":43,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,17]],"date-time":"2025-11-17T00:00:00Z","timestamp":1763337600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["U01CA244100"],"award-info":[{"award-number":["U01CA244100"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Blinatumomab, a CD3\/CD19 bispecific T-cell engager (TCE), is effective in relapsed\/refractory (R\/R) B-cell acute lymphoblastic leukemia (B-ALL), but responses may be limited by T cell exhaustion. A recent preclinical study found that incorporating a 7-day treatment-free interval (TFI) into TCE exposure reduced T cell exhaustion and improved subsequent antitumor activity compared with continuous exposure.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Based on this preclinical study, we developed an agent-based model (ABM) to evaluate alternative TFIs by simulating T cell\u2013tumor cell interactions, in which TCE availability influences T cell activation, tumor-cell killing, and T cell exhaustion. To reproduce the experimental design, simulations included a 14-day in vitro conditioning phase with varying TCE schedules, followed by an adoptive cell transfer (ACT) phase in which post-transfer tumor control was evaluated.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Our ABM reproduced the experimentally observed reduction in CD8\u207a T cell exhaustion following a 7-day TFI versus continuous TCE exposure (CONT). Among tested TFIs, the 7-day TFI produced the lowest exhaustion (p_adj = 3.52 \u00d7 10\u207b\u00b9\u2077 vs. CONT) and lowest tumor burden (p_adj = 1.93 \u00d7 10\u207b\u00b9\u2076 vs. CONT). The 5-day TFI was the only other tested schedule to significantly reduce tumor burden relative to CONT. Shifting the 7-day TFI from the second to the first week of conditioning altered T cell phenotype at harvest but not tumor control during the ACT phase (p_adj = 0.87). Limiting TCE exposure during the in vitro phase to one day further reduced tumor burden (all p_adj &lt; 5 \u00d7 10\u207b\u00b9\u2077), with exposure on the first day outperforming exposure immediately before T cell harvest (p_adj = 3.6 \u00d7 10\u207b\u00b9\u2070).<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The benefit of TFIs depends on interval duration and schedule organization. Our findings also highlight a potential limitation of current preclinical TFI testing. Because tumor cells are replenished rather than allowed to grow during in vitro conditioning, prolonged TFIs carry no penalty from tumor growth. Models that apply TFIs directly to growing tumors are needed to determine whether reduced T cell exhaustion outweighs tumor progression during treatment withdrawal.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Author Summary<\/jats:title>\n                  <jats:p>T cell engagers help T cells recognize and attack tumor cells. Although these treatments can help control tumors, they may also overstimulate T cells, causing them to become exhausted and lose function. Temporarily interrupting treatment could give T cells time to recover, but testing the many possible ways to schedule these treatment breaks experimentally would be difficult. We therefore developed a computer model that simulates T cells and tumor cells to investigate how the duration and timing of treatment breaks influence T cell exhaustion and subsequent tumor control. Our model reproduced experimental findings showing that treatment breaks can preserve T cell function. We then used the model to compare additional schedules. In general, longer treatment breaks preserved T cell function better than several shorter breaks. We also examined whether the placement of the break within the treatment schedule mattered and found that moving the same break to an earlier point did not affect subsequent tumor control. Our findings show how computer models can help compare treatment schedules and reveal that more realistic models and experimental testing are needed.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.11.17.688873","type":"posted-content","created":{"date-parts":[[2025,11,18]],"date-time":"2025-11-18T05:10:46Z","timestamp":1763442646000},"source":"Crossref","is-referenced-by-count":0,"title":["Treatment-free interval duration during T-cell engager conditioning shapes simulated T-cell exhaustion and post-transfer tumor control"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-5100-1590","authenticated-orcid":false,"given":"Nina","family":"Obertopp","sequence":"first","affiliation":[{"name":"Cancer Biology Ph.D. Program, University of South Florida, Tampa, Florida, USA"},{"name":"Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1231-7297","authenticated-orcid":false,"given":"Matthew","family":"Froid","sequence":"additional","affiliation":[{"name":"Cancer Biology Ph.D. Program, University of South Florida, Tampa, Florida, USA"},{"name":"Department of Integrated Mathematical Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7785-3034","authenticated-orcid":false,"given":"Shari","family":"Pilon-Thomas","sequence":"additional","affiliation":[{"name":"Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8527-0776","authenticated-orcid":false,"given":"David","family":"Basanta","sequence":"additional","affiliation":[{"name":"Department of Integrated Mathematical Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026100504500577000_2025.11.17.688873v2.1","doi-asserted-by":"publisher","DOI":"10.4084\/MJHID.2024.070"},{"key":"2026100504500577000_2025.11.17.688873v2.2","doi-asserted-by":"crossref","first-page":"5888","DOI":"10.1182\/blood-2024-208221","article-title":"A Systematic Review on Efficacy and Safety Profile of Blinatumomab in Relapsed or Refractory B-Cell Precursor Acute Lymphoblastic Leukemia (R\/R BCP-ALL) in Children and Young Adults","volume":"144","year":"2024","journal-title":"Blood"},{"key":"2026100504500577000_2025.11.17.688873v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2023.02.008"},{"key":"2026100504500577000_2025.11.17.688873v2.4","doi-asserted-by":"publisher","DOI":"10.1126\/science.aba1786"},{"key":"2026100504500577000_2025.11.17.688873v2.5","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2022015956"},{"key":"2026100504500577000_2025.11.17.688873v2.6","unstructured":"BLINCYTO\u00ae (blinatumomab) Dosing. 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