Kayla Kulhanek

All Publications

• IL-9 as a naturally orthogonal cytokine with optimal JAK/STAT signaling for engineered T cell therapy. Immunity Jiang, H., Limsuwannarot, S., Kulhanek, K. R., Pal, A., Labiad, O., Rysavy, L. W., Wong, A., Su, L., Cavender, S., Soro, J., Testa, S., Ogana, H., Waghray, D., Tao, P., Jude, K. M., Seet, C. S., Crooks, G. M., Moding, E. J., Garcia, K. C., Kalbasi, A. 2025

  Abstract

  Cytokines and their receptors enable precise tuning of T cell function. Leveraging this biology holds tremendous promise for optimizing antitumor immunity. Arming T cells with a synthetically orthogonal interleukin (IL)-9 receptor (o9R), for instance, permits facile engraftment and potent anti-tumor functions. Exploiting the paucity of wild-type IL-9R expression and the safety of high doses of IL-9, here, we showed that, compared with o9R, T cells engineered with wild-type IL-9R exhibited superior tissue infiltration, stemness, and anti-tumor activity. These qualities were consistent with a stronger Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signal, which included canonically IL-12-driven STAT4 in addition to STAT1/3/5. IL-9R T cells were exquisitely sensitive to perturbations of proximal signaling, including structure-guided attenuation, amplification, and rebalancing of JAK/STAT signals. Biased IL-9R mutants showed that STAT1 acts as a rheostat between stem-like and effector states. In summary, we identify IL-9/IL-9R as a naturally orthogonal cytokine-receptor pair with an optimal JAK/STAT signaling profile for engineered T cell therapy.

  View details for DOI 10.1016/j.immuni.2025.10.024

  View details for PubMedID 41274289

  View details for PubMedCentralID PMC12643178

• Bariatric Surgery: Improving Access without Compromising Outcomes. Obesity surgery Chinn, J. O., Bartoletti, S., Shen, J. X., Kulhanek, K. R., Shacker, M., Zak, Y., Azagury, D. E., Esquivel, M. M. 2025

  Abstract

  Bariatric surgery pre-operative workup mandates many multidisciplinary visits demanding patient's commitment in time and travel. Due to the COVID pandemic, our bariatric clinic transitioned to a telemedicine model. The objective of this work is to determine the impact of this shift.Our population is adults who underwent gastric bypass or sleeve gastrectomy at a single hospital between 2018 and 2022. We analyzed the impact of telemedicine on demographics, days from consultation to surgery, length of hospital stay, type of procedures, BMI, and complications. Statistical analyses were conducted using χ2 tests for categorical variables and t-test for continuous variables as well as logistic regression.Of 794 patients, 71.3% received in-person care while 28.7% received telemedicine. There were no differences in race, ethnicity, or gender. The average duration from consultation to surgery was longer (p < 0.001) for in-person (551.1 days) compared to telemedicine (375.8 days). The pre-operative BMI was higher for the in-person cohort versus telemedicine (45.5; SD 8.0 vs. 43.9; SD 6.7; p < 0.001), but there was no difference in percent weight loss at 12 months. The average length of stay was longer in the in-person group compared to telemedicine (1.9 vs 1.2 days). There was no difference in readmission or emergency department visit rates.Despite being the best treatment for severe obesity, bariatric surgery remains underutilized. When comparing telemedicine to an in-person model, we found shorter time to surgery, shorter length of stay, and similar outcomes with no difference in weight loss or complications. Telemedicine may be a safe and useful way to improve access.

  View details for DOI 10.1007/s11695-025-08222-8

  View details for PubMedID 40921958

  View details for PubMedCentralID 11325826

• The Impact of Socioeconomic Disadvantage on Metabolic and Bariatric Surgical Outcomes using Area Deprivation Index. Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract Chinn, J. O., Kulhanek, K. R., Shen, J. X., Shacker, M., Zak, Y., Azagury, D. E., Esquivel, M. M. 2025: 102136

  View details for DOI 10.1016/j.gassur.2025.102136

  View details for PubMedID 40582560

• IL-9 as a naturally orthogonal cytokine with optimal JAK/STAT signaling for engineered T cell therapy. bioRxiv : the preprint server for biology Jiang, H., Limsuwannarot, S., Kulhanek, K. R., Pal, A., Rysavy, L. W., Su, L., Labiad, O., Testa, S., Ogana, H., Waghray, D., Tao, P., Jude, K. M., Seet, C. S., Crooks, G. M., Moding, E. J., Garcia, K. C., Kalbasi, A. 2025

  Abstract

  Arming T cells with a synthetically orthogonal IL-9 receptor (o9R) permits facile engraftment and potent anti-tumor functions. We considered whether the paucity of natural IL-9R expression could be exploited for T cell immunotherapy given that, in mice, high doses of IL-9 were well-tolerated without discernible immune modulation. Compared to o9R, T cells engineered with IL-9R exhibit superior tissue infiltration, stemness, and anti-tumor activity. These qualities are consistent with a stronger JAK/STAT signal, which in addition to STAT1/3/5, unexpectedly includes STAT4 (canonically associated with IL-12 but not common γ-chain cytokines). IL-9R T cells are exquisitely sensitive to perturbations of proximal signaling, including structure-guided attenuation, amplification, and rebalancing of JAK/STAT signals. Biased IL-9R mutants uncover STAT1 as a rheostat between proliferative stem-like and terminally differentiated effector states. In summary, we identify native IL-9/IL-9R as a natural cytokine-receptor pair with near-orthogonal qualities and an optimal JAK/STAT signaling profile for engineered T cell therapy.

  View details for DOI 10.1101/2025.01.15.633105

  View details for PubMedID 39868284

  View details for PubMedCentralID PMC11760723

• The Impact of Area Deprivation Index on Bariatric Surgical Outcomes Chinn, J., Adams, M., Kulhanek, K. R., Shen, J. X., Tennakoon, L., Bartoletti, S., Azagury, D. E., Esquivel, M. LIPPINCOTT WILLIAMS & WILKINS. 2024: S44

  View details for Web of Science ID 001348680700063

• Targeted Bias: The Next Swing at IL2 Therapy. Cancer discovery Kulhanek, K. R., Kalbasi, A. 2024; 14 (7): 1145-1146

  Abstract

  Despite its long history of toxicity and limited efficacy, IL2 has re-entered the clinic as a companion to the recently FDA-approved tumor infiltrating lymphocyte therapy. In back-to-back articles, Moynihan and Kaptein introduce a new fusion protein that delivers a biased IL2 mutein to CD8 T cells. See related article by Moynihan et al., p. 1206 (6). See related article by Kaptein et al., p. 1226 (7).

  View details for DOI 10.1158/2159-8290.CD-24-0558

  View details for PubMedID 38946323

• SARS-CoV-2 infection of airway organoids reveals conserved use of Tetraspanin-8 by Ancestral, Delta, and Omicron variants. Stem cell reports Hysenaj, L., Little, S., Kulhanek, K., Magnen, M., Bahl, K., Gbenedio, O. M., Prinz, M., Rodriguez, L., Andersen, C., Rao, A. A., Shen, A., Lone, J. C., Lupin-Jimenez, L. C., Bonser, L. R., Serwas, N. K., Mick, E., Khalid, M. M., Taha, T. Y., Kumar, R., Li, J. Z., Ding, V. W., Matsumoto, S., Maishan, M., Sreekumar, B., Simoneau, C., Nazarenko, I., Tomlinson, M. G., Khan, K., von Gottberg, A., Sigal, A., Looney, M. R., Fragiadakis, G. K., Jablons, D. M., Langelier, C. R., Matthay, M., Krummel, M., Erle, D. J., Combes, A. J., Sil, A., Ott, M., Kratz, J. R., Roose, J. P. 2023; 18 (3): 636-653

  Abstract

  Ancestral SARS coronavirus-2 (SARS-CoV-2) and variants of concern (VOC) caused a global pandemic with a spectrum of disease severity. The mechanistic explaining variations related to airway epithelium are relatively understudied. Here, we biobanked airway organoids (AO) by preserving stem cell function. We optimized viral infection with H1N1/PR8 and comprehensively characterized epithelial responses to SARS-CoV-2 infection in phenotypically stable AO from 20 different subjects. We discovered Tetraspanin-8 (TSPAN8) as a facilitator of SARS-CoV-2 infection. TSPAN8 facilitates SARS-CoV-2 infection rates independently of ACE2-Spike interaction. In head-to-head comparisons with Ancestral SARS-CoV-2, Delta and Omicron VOC displayed lower overall infection rates of AO but triggered changes in epithelial response. All variants shared highest tropism for ciliated and goblet cells. TSPAN8-blocking antibodies diminish SARS-CoV-2 infection and may spur novel avenues for COVID-19 therapy.

  View details for DOI 10.1016/j.stemcr.2023.01.011

  View details for PubMedID 36827975

  View details for PubMedCentralID PMC9948283

• Dysregulated RASGRP1 expression through RUNX1 mediated transcription promotes autoimmunity. European journal of immunology Baars, M. J., Douma, T., Simeonov, D. R., Myers, D. R., Kulhanek, K., Banerjee, S., Zwakenberg, S., Baltissen, M. P., Amini, M., de Roock, S., van Wijk, F., Vermeulen, M., Marson, A., Roose, J. P., Vercoulen, Y. 2021; 51 (2): 471-482

  Abstract

  RasGRP1 is a Ras guanine nucleotide exchange factor, and an essential regulator of lymphocyte receptor signaling. In mice, Rasgrp1 deletion results in defective T lymphocyte development. RASGRP1-deficient patients suffer from immune deficiency, and the RASGRP1 gene has been linked to autoimmunity. However, how RasGRP1 levels are regulated, and if RasGRP1 dosage alterations contribute to autoimmunity remains unknown. We demonstrate that diminished Rasgrp1 expression caused defective T lymphocyte selection in C57BL/6 mice, and that the severity of inflammatory disease inversely correlates with Rasgrp1 expression levels. In patients with autoimmunity, active inflammation correlated with decreased RASGRP1 levels in CD4+ T cells. By analyzing H3K27 acetylation profiles in human T cells, we identified a RASGRP1 enhancer that harbors autoimmunity-associated SNPs. CRISPR-Cas9 disruption of this enhancer caused lower RasGRP1 expression, and decreased binding of RUNX1 and CBFB transcription factors. Analyzing patients with autoimmunity, we detected reduced RUNX1 expression in CD4+ T cells. Lastly, we mechanistically link RUNX1 to transcriptional regulation of RASGRP1 to reveal a key circuit regulating RasGRP1 expression, which is vital to prevent inflammatory disease.

  View details for DOI 10.1002/eji.201948451

  View details for PubMedID 33065764

  View details for PubMedCentralID PMC7894479

• Protocol for Barcoding T Cells Combined with Timed Stimulations. STAR protocols Kulhanek, K. R., Myers, D. R., Ksionda, O., Vercoulen, Y., Romero-Moya, D., Roose, J. P. 2020; 1 (2): 100067

  Abstract

  Stimulation of naive T lymphocytes via the T cell receptor (TCR) induces distinct phosphorylation patterns that can be used to explore various signaling pathways within the cell. This protocol can be used to characterize different perturbations to the signaling pathways and the variations in time of stimulation. Here, we provide a method of barcoding and consolidating a maximum of 24 different sample conditions using two florescent dyes. This single sample for phospho-staining and flow cytometry saves time and reagents. For complete details on the use and execution of this protocol, please refer to Krutzik and Nolan (2006), Krutzik et al. (2012), Vercoulen et al. (2017), Ksionda et al. (2018), and Myers et al. (2019).

  View details for DOI 10.1016/j.xpro.2020.100067

  View details for PubMedID 33111105

  View details for PubMedCentralID PMC7580249

• Cryo-EM structure of a dimeric B-Raf:14-3-3 complex reveals asymmetry in the active sites of B-Raf kinases. Science (New York, N.Y.) Kondo, Y., Ognjenović, J., Banerjee, S., Karandur, D., Merk, A., Kulhanek, K., Wong, K., Roose, J. P., Subramaniam, S., Kuriyan, J. 2019; 366 (6461): 109-115

  Abstract

  Raf kinases are important cancer drug targets. Paradoxically, many B-Raf inhibitors induce the activation of Raf kinases. Cryo-electron microscopy structural analysis of a phosphorylated B-Raf kinase domain dimer in complex with dimeric 14-3-3, at a resolution of ~3.9 angstroms, shows an asymmetric arrangement in which one kinase is in a canonical "active" conformation. The distal segment of the C-terminal tail of this kinase interacts with, and blocks, the active site of the cognate kinase in this asymmetric arrangement. Deletion of the C-terminal segment reduces Raf activity. The unexpected asymmetric quaternary architecture illustrates how the paradoxical activation of Raf by kinase inhibitors reflects an innate mechanism, with 14-3-3 facilitating inhibition of one kinase while maintaining activity of the other. Conformational modulation of these contacts may provide new opportunities for Raf inhibitor development.

  View details for DOI 10.1126/science.aay0543

  View details for PubMedID 31604311

  View details for PubMedCentralID PMC7007921

• Alternative ZAP70-p38 signals prime a classical p38 pathway through LAT and SOS to support regulatory T cell differentiation. Science signaling Jun, J. E., Kulhanek, K. R., Chen, H., Chakraborty, A., Roose, J. P. 2019; 12 (591)

  Abstract

  T cell receptor (TCR) stimulation activates diverse kinase pathways, which include the mitogen-activated protein kinases (MAPKs) ERK and p38, the phosphoinositide 3-kinases (PI3Ks), and the kinase mTOR. Although TCR stimulation activates the p38 pathway through a "classical" MAPK cascade that is mediated by the adaptor protein LAT, it also stimulates an "alternative" pathway in which p38 is activated by the kinase ZAP70. Here, we used dual-parameter, phosphoflow cytometry and in silico computation to investigate how both classical and alternative p38 pathways contribute to T cell activation. We found that basal ZAP70 activation in resting T cell lines reduced the threshold ("primed") TCR-stimulated activation of the classical p38 pathway. Classical p38 signals were reduced after T cell-specific deletion of the guanine nucleotide exchange factors Sos1 and Sos2, which are essential LAT signalosome components. As a consequence of Sos1/2 deficiency, production of the cytokine IL-2 was impaired, differentiation into regulatory T cells was reduced, and the autoimmune disease EAE was exacerbated in mice. These data suggest that the classical and alternative p38 activation pathways exist to generate immune balance.

  View details for DOI 10.1126/scisignal.aao0736

  View details for PubMedID 31337738

  View details for PubMedCentralID PMC7340343