{"id":{"repo_id":"tuebingen","oai_identifier":"oai:publikationen.uni-tuebingen.de:10900/151271"},"canonical_url":"https://search.dev.ndltd.org/etd/tuebingen/oai:publikationen.uni-tuebingen.de:10900/151271","repository":{"repo_id":"tuebingen","name":"Universität Tübingen","base_url":"https://publikationen.uni-tuebingen.de/oai/request"},"display":{"title":"Computational Design and Optimization of G-CSFR Modulators","abstract":"Considering the indispensable role played by proteins in maintaining vital life processes, it is not surprising that proteins are linked to a broad spectrum of diseases. Conversely, proteins can be leveraged for effective therapeutic interventions. For example, secreted growth factors known as cytokines have emerged as promising candidates for protein-based therapeutics, primarily due to their potent immunomodulatory properties. An integral element of the innate immune system consists of neutrophils, the maturation of which is coordinated by a hematopoietic subprocess known as granulopoiesis. This process requires a complex interplay among various cytokines and their corresponding receptor molecules. A critical player in these interactions is the granulocyte-colony stimulating factor receptor (G-CSFR), which is activated by its native ligand, G-CSF. Unlike most other cytokines, G-CSF has found clinical use in its native form due to its favorable safety profile and its ability to increase the number of neutrophils in the blood. However, G-CSF application is restricted by its stability, production cost and native activity on G-CSFR. Additionally, there is very limited knowledge regarding non-native G-CSFR modulators, which could be key not only for understanding G-CSFR related diseases, but also for the development of innovative therapeutic applications. In order to unlock the untapped potential of the clinically significant G-CSFR beyond its native activity, my objective was to employ protein design techniques to craft novel ligands capable of modulating G-CSFR activity. In addition to customizing receptor activity, these designs offer enhanced stability and more efficient production compared to their native counterpart G-CSF. To this end, I utilized a recently developed hyper-thermostable de novo designed GCSFR binding module and optimized it with in silico and in vitro high-throughput methods to obtain a broad spectrum of variants with enhanced binding affinity. I demonstrate that these enhanced binding modules can be utilized to generate GCSFR agonists that achieve G-CSF activity in cell-based assays and can also be used to create ligands capable of modulating G-CSFR activity by tuning receptor geometry. These ligands featured fine-tuned intracellular signaling, transcriptomic activity, and primary stem cell differentiation and exhibit in vivo activity in zebrafish and mouse models. In addition to designing agonists, I show that these enhanced binding modules can be used to generate competitive G-CSFR antagonists with nanomolar inhibitory activity. To the best of my knowledge this work is the first demonstration that G-CSFR activity can be tuned by the design of ligands inducing non-native receptor geometries. Additionally, this study presents an array of binding modules with diverse affinities to G-CSFR, along with the newly designed ligands, providing the foundational components for systematic investigation of G-CSFR activity modulation. These findings hold significant promise for advancing the development of innovative protein therapeutics.","abstract_html":"Considering the indispensable role played by proteins in maintaining vital life processes, it is not surprising that proteins are linked to a broad spectrum of diseases. Conversely, proteins can be leveraged for effective therapeutic interventions. For example, secreted growth factors known as cytokines have emerged as promising candidates for protein-based therapeutics, primarily due to their potent immunomodulatory properties. An integral element of the innate immune system consists of neutrophils, the maturation of which is coordinated by a hematopoietic subprocess known as granulopoiesis. This process requires a complex interplay among various cytokines and their corresponding receptor molecules. A critical player in these interactions is the granulocyte-colony stimulating factor receptor (G-CSFR), which is activated by its native ligand, G-CSF. Unlike most other cytokines, G-CSF has found clinical use in its native form due to its favorable safety profile and its ability to increase the number of neutrophils in the blood. However, G-CSF application is restricted by its stability, production cost and native activity on G-CSFR. Additionally, there is very limited knowledge regarding non-native G-CSFR modulators, which could be key not only for understanding G-CSFR related diseases, but also for the development of innovative therapeutic applications. In order to unlock the untapped potential of the clinically significant G-CSFR beyond its native activity, my objective was to employ protein design techniques to craft novel ligands capable of modulating G-CSFR activity. In addition to customizing receptor activity, these designs offer enhanced stability and more efficient production compared to their native counterpart G-CSF. To this end, I utilized a recently developed hyper-thermostable de novo designed GCSFR binding module and optimized it with in silico and in vitro high-throughput methods to obtain a broad spectrum of variants with enhanced binding affinity. I demonstrate that these enhanced binding modules can be utilized to generate GCSFR agonists that achieve G-CSF activity in cell-based assays and can also be used to create ligands capable of modulating G-CSFR activity by tuning receptor geometry. These ligands featured fine-tuned intracellular signaling, transcriptomic activity, and primary stem cell differentiation and exhibit in vivo activity in zebrafish and mouse models. In addition to designing agonists, I show that these enhanced binding modules can be used to generate competitive G-CSFR antagonists with nanomolar inhibitory activity. To the best of my knowledge this work is the first demonstration that G-CSFR activity can be tuned by the design of ligands inducing non-native receptor geometries. Additionally, this study presents an array of binding modules with diverse affinities to G-CSFR, along with the newly designed ligands, providing the foundational components for systematic investigation of G-CSFR activity modulation. These findings hold significant promise for advancing the development of innovative protein therapeutics.","abstract_has_math":false,"creators":["Ullrich, Timo"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2026,"date_issued":"2026-01-30","date_published":"2026-01-30","updated_at":"2026-08-21T22:21:56Z","subjects":[],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["hdl:10900/151271"],"render_values":[{"text":"hdl:10900/151271","href":null,"code":true}]}]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"source_record":{"url":"https://publikationen.uni-tuebingen.de/oai/request?verb=GetRecord&metadataPrefix=mets&identifier=oai%3Apublikationen.uni-tuebingen.de%3A10900%2F151271","prefix":"mets"},"metadata_groups":[{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2026-01-30"]},{"key":"dc:type","label":"Dc Type","values":["PhDThesis"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["hdl:10900/151271"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.other","label":"Dc Description Other","values":["Considering the indispensable role played by proteins in maintaining vital life processes, it is not surprising that proteins are linked to a broad spectrum of diseases. Conversely, proteins can be leveraged for effective therapeutic interventions. For example, secreted growth factors known as cytokines have emerged as promising candidates for protein-based therapeutics, primarily due to their potent immunomodulatory properties. An integral element of the innate immune system consists of neutrophils, the maturation of which is coordinated by a hematopoietic subprocess known as granulopoiesis. This process requires a complex interplay among various cytokines and their corresponding receptor molecules. A critical player in these interactions is the granulocyte-colony stimulating factor receptor (G-CSFR), which is activated by its native ligand, G-CSF. Unlike most other cytokines, G-CSF has found clinical use in its native form due to its favorable safety profile and its ability to increase the number of neutrophils in the blood. However, G-CSF application is restricted by its stability, production cost and native activity on G-CSFR. Additionally, there is very limited knowledge regarding non-native G-CSFR modulators, which could be key not only for understanding G-CSFR related diseases, but also for the development of innovative therapeutic applications. In order to unlock the untapped potential of the clinically significant G-CSFR beyond its native activity, my objective was to employ protein design techniques to craft novel ligands capable of modulating G-CSFR activity. In addition to customizing receptor activity, these designs offer enhanced stability and more efficient production compared to their native counterpart G-CSF. To this end, I utilized a recently developed hyper-thermostable de novo designed GCSFR binding module and optimized it with in silico and in vitro high-throughput methods to obtain a broad spectrum of variants with enhanced binding affinity. I demonstrate that these enhanced binding modules can be utilized to generate GCSFR agonists that achieve G-CSF activity in cell-based assays and can also be used to create ligands capable of modulating G-CSFR activity by tuning receptor geometry. These ligands featured fine-tuned intracellular signaling, transcriptomic activity, and primary stem cell differentiation and exhibit in vivo activity in zebrafish and mouse models. In addition to designing agonists, I show that these enhanced binding modules can be used to generate competitive G-CSFR antagonists with nanomolar inhibitory activity. To the best of my knowledge this work is the first demonstration that G-CSFR activity can be tuned by the design of ligands inducing non-native receptor geometries. Additionally, this study presents an array of binding modules with diverse affinities to G-CSFR, along with the newly designed ligands, providing the foundational components for systematic investigation of G-CSFR activity modulation. These findings hold significant promise for advancing the development of innovative protein therapeutics."]},{"key":"dc:title","label":"Title","values":["Computational Design and Optimization of G-CSFR Modulators"]}]}],"canonical_facts":{"dc:date.issued":["2026-01-30"],"dc:description.other":["Considering the indispensable role played by proteins in maintaining vital life processes, it is not surprising that proteins are linked to a broad spectrum of diseases. Conversely, proteins can be leveraged for effective therapeutic interventions. For example, secreted growth factors known as cytokines have emerged as promising candidates for protein-based therapeutics, primarily due to their potent immunomodulatory properties. An integral element of the innate immune system consists of neutrophils, the maturation of which is coordinated by a hematopoietic subprocess known as granulopoiesis. This process requires a complex interplay among various cytokines and their corresponding receptor molecules. A critical player in these interactions is the granulocyte-colony stimulating factor receptor (G-CSFR), which is activated by its native ligand, G-CSF. Unlike most other cytokines, G-CSF has found clinical use in its native form due to its favorable safety profile and its ability to increase the number of neutrophils in the blood. However, G-CSF application is restricted by its stability, production cost and native activity on G-CSFR. Additionally, there is very limited knowledge regarding non-native G-CSFR modulators, which could be key not only for understanding G-CSFR related diseases, but also for the development of innovative therapeutic applications. In order to unlock the untapped potential of the clinically significant G-CSFR beyond its native activity, my objective was to employ protein design techniques to craft novel ligands capable of modulating G-CSFR activity. In addition to customizing receptor activity, these designs offer enhanced stability and more efficient production compared to their native counterpart G-CSF. To this end, I utilized a recently developed hyper-thermostable de novo designed GCSFR binding module and optimized it with in silico and in vitro high-throughput methods to obtain a broad spectrum of variants with enhanced binding affinity. I demonstrate that these enhanced binding modules can be utilized to generate GCSFR agonists that achieve G-CSF activity in cell-based assays and can also be used to create ligands capable of modulating G-CSFR activity by tuning receptor geometry. These ligands featured fine-tuned intracellular signaling, transcriptomic activity, and primary stem cell differentiation and exhibit in vivo activity in zebrafish and mouse models. In addition to designing agonists, I show that these enhanced binding modules can be used to generate competitive G-CSFR antagonists with nanomolar inhibitory activity. To the best of my knowledge this work is the first demonstration that G-CSFR activity can be tuned by the design of ligands inducing non-native receptor geometries. Additionally, this study presents an array of binding modules with diverse affinities to G-CSFR, along with the newly designed ligands, providing the foundational components for systematic investigation of G-CSFR activity modulation. These findings hold significant promise for advancing the development of innovative protein therapeutics."],"dc:identifier":["hdl:10900/151271"],"dc:title":["Computational Design and Optimization of G-CSFR Modulators"],"dc:type":["PhDThesis"]},"updated_at":"2026-08-21T22:21:56Z"}