Paul Kulesa, PhD

Professor of Pediatrics, University of Missouri-Kansas City School of Medicine

Full Biography

In February 2024, internationally recognized developmental neuroscientist Paul Kulesa, PhD, joined Children’s Mercy Research Institute (CMRI) in the Division of Developmental and Behavioral Health. His group studies how errors in cell migration and cell differentiation during early development lead to birth defects and disease. By closely collaborating with surgeons and clinicians, his goal is to leverage new discoveries in the laboratory toward novel tissue repair and therapeutic treatments that improve infant survival and quality of life.

His research focuses on neural crest cell development in the embryo and how these cells migrate long distances and differentiate into specific cell types (bone, cartilage and neurons). Neural crest cells travel throughout the embryo to build the face, organs and peripheral nervous system. Dr. Kulesa and his group plan to explain how developmental signals regulate neural crest cell behaviors. They will use this knowledge to design strategies to repair craniofacial birth defects and prevent the transition from normal development to disease, such as in the pediatric cancer, neuroblastoma. 

The Kulesa Lab: Building for innovation

The Kulesa Lab includes a multidisciplinary team equipped with advanced imaging technology. The new imaging capabilities will allow his team to apply innovative techniques to study cell-neighbor and cell-microenvironment interactions, critical aspects of collective migration. 

Team

The Kulesa Lab team includes Jennifer Kasemeier, PhD, Sam Mellentine, PhD, Noelle Parisi Dahl, MS, and Daniel Kremer, BS. Dr. Kulesa plans to add an additional data/experimental scientist to complete a multi-disciplinary group.  His team also works closely with other CMRI teams, including those with expertise in research informatics, genomics, bioinformatics and pathology. The team also collaborates with clinicians throughout CMKC as well as physicians in Boston and scientists in Oxford England with expertise in mathematical modeling and deep learning. In the future, Dr. Kulesa envisions bench scientists and clinical residents training and interacting side-by-side, using state-of-the-art methods to solve complex biomedical challenges. 

State of the art imaging

In spring 2024, CMRI received its first spinning disk confocal microscope. Located near the Kulesa Lab and in general use at CMRI, this microscope has a multi-laser line module that allows users to view fluorescently-labeled cell and tissue samples with up to seven different colors instead of the typical one or two, dramatically accelerating the analysis of how cell and tissue components may interact. The multi-laser line module (Ziva light engine) is one of the first of its kind in North America. The microscope is already in use by multiple teams throughout CMRI. “People are excited by the high resolution and ability to more completely visualize cell and tissue architectures in the same sample,” said Dr. Kulesa. 

This fall, the lab will acquire a second confocal microscope with an environmental chamber around the microscope stage that will allow for visualization of dynamic processes in living cells and tissues. Also, a module on the microscope is capable of super-resolution microscopy – allowing the observation of neighboring cellular components that are typically too close to resolve. “We will now be able to see subcellular structures, how they are organized within the cell, and how these subcellular components react to changes when we manipulate particular genes,” said Dr. Kulesa.

Patient images combined with spatial gene expression data may help to better predict the disease progression and suggest new therapeutic targets.

Over the next year, Dr. Kulesa plans to add a two-photon microscope, allowing scientists to see deeper into tissue samples with less photodamage, and a compound microscope with a high resolution camera and moving stage to visualize and study the rapidly growing field of patient-derived organoids. 

Innovative techniques

In addition to the new imaging technology, the Kulesa Lab brings its expertise in spatial transcriptomics (a means to map gene expression with cell position) and multiplexed fluorescence in-situ hybridization (a technique to view gene expression levels in a cell). This will allow scientists and clinicians the ability to measure and validate gene expression patterns in intact tissue samples without requiring cells to be isolated – preserving cell-neighbor relationships and the local microenvironment. This will help researchers better understand the role of complex cell and tissue interactions. “Previously, we disconnected cells and lost their spatial context, which we know is important to the systems biology within the tissue architecture,” said Dr. Kulesa. “Now, there’s the potential to measure how gene expression changes drive local events within the tissue and subsequent function of the biological process.” 

Dr. Kulesa recently co-authored a paper on this technique for the journal Developmental Dynamics.

His team will also employ multiplexing techniques to study RNA/protein in tissue samples. Multiplexing allows researchers to look at multiple gene targets in the same tissue sample. Dr. Kulesa said that this can explain the relationships between multiple cells. “This allows one to correlate the expression of a particular gene with another gene because many of these genes function cooperatively. They work in signaling pathways that might be shared or overlap,” he said. It also speeds up the process to analyze multiple gene targets derived from large-scale genome sequencing or a standard set of biomarkers analyzed by pathologists. 

Neuroblastoma project

Dr. Kulesa will use the new imaging equipment and innovative techniques in a multi-team project to study neuroblastoma cancer. Neuroblastoma is the most common cancer in infants ages zero to 18 months, yet it remains unclear how neuroblastoma is initiated and progresses by errors in neural crest cell differentiation and peripheral nervous system development.

“There are mistakes in developmental signals during gestation that appear to give rise to a neuroblastoma tumor which then progresses during the early life of a newborn,” said Dr. Kulesa. “This critical time period of zero to 18 months is a window where we believe there's fertile ground to explore the developmental origins of neuroblastoma, and if we can understand this, we may be able to better predict and treat neuroblastoma progression.”

For this project, Dr. Kulesa and his colleagues will analyze human neuroblastoma patient images and tissue samples, profile the gene expression patterns in those samples, and use this knowledge to predict and suggest treatments to control the disease progression in a personalized manner. Previously, Dr. Kulesa co-authored a paper in the journal Developmental Biology on screening for the most aggressive neuroblastoma cells in patient-derived primary cells. 

Impact to treatment

Dr. Kulesa explains how this may impact treatment for patients. “Patient images combined with spatial gene expression data may help to better predict the disease progression and suggest basic science experiemnst to derive new therapeutic targets,” said Dr. Kulesa. “This knowledge may provide a clearer picture for pediatric oncologists and surgeons to decide whether to begin aggressive chemotherapy or radiation or wait and see whether the tumor will either fully differentiate into a benign mass or shrink to a small mass without treatment – a striking characteristic of some aggressive neuroblastoma tumors, called spontaneous regression. This knowledge may also provide insights that accelerate clinical studies to inhibit cell proliferation, promote cell differentiation and induce tumor regression in relapsed patients.” 

Project team

The project’s team will be a blend of scientists and clinicians. “On the science side, it will include people with expertise in developmental neuroscience, research informatics, genomics, and bioinformatics. On the clinical side, the team will include experts in pediatric surgery, oncology, and pathology,” said Dr. Kulesa. “Our goal is to build a convergent biosciences environment with lots of crosstalk between basic science and clinical work.”

Recent awards

In October 2024, Dr. Kulesa received a $1.9 million R01 award from the National Institutes of Health (NIH) for his project, “Investigating the relationship between sympathetic nervous system development and neuroblastoma.” Dr. Kulesa hopes that study results will lead to translational research to repair peripheral nerve connections and inform drug design efforts for patients with neuroblastoma. He also received an NIH R21 grant transfer for his study, “A novel platform to enhance single cell interrogation of nervous system development.” The goal of this study is to advance methods in spatial transcriptomics and multiplexing.  Dr. Kasemeier is a co-investigator on both studies. In December 2024, Dr. Kulesa received an internal Children’s Mercy award to further his neuroblastoma research.

Future goals

Through collaboration between scientists and clinicians, Dr. Kulesa hopes to build a center for neuroblastoma research. “We want to pull together the technology and the clinical side to predict whether an individual infant’s neuroblastoma tumor will regress, a particularly exciting research area that is understudied,” he said. “If we can understand the mechanistic basis for neuroblastoma regression, then we may develop a treatment protocol to induce regression.”

Dr. Kulesa also plans to collaborate with other teams at CMKC on a variety of conditions with developmental origins. “Once we have the innovative technologies infrastructure in place, there are many other areas to collaborate with,” he said. “There are links to other developmental and behavioral health questions that have developmental origins related to the neural crest cells we study.”