About

Donita C. Brady, Ph.D., is a Presidential Professor at the University of Pennsylvania's Perelman School of Medicine and a faculty member in the Department of Cancer Biology. Her research interests lie at the intersection of cancer biology, signal transduction, and metal homeostasis. Her laboratory explores how metals contribute to cancer growth by supporting cellular processes involving DNA, RNA, and proteins, focusing on how cells maintain metal balance and respond to fluctuations, particularly in the context of tissue health, development, and cancer. Additionally, her team investigates protein function changes in cancer beyond genetic differences, utilizing platforms like PEAR to identify drug targets and improve precision oncology. Brady's work aims to uncover new insights into metal-driven cellular mechanisms and develop targeted therapies for cancer.

Research topics

• Biology
• Social Science
• Sociology
• Cell biology
• Engineering ethics
• Environmental ethics
• Chemistry
• Genetics
• Gender studies
• Biochemistry
• Cancer research

Selected publications

• Reduction in Hepatic Phosphatidylcholine Biosynthesis Promotes MASH Through Copper Deficiency

  bioRxiv (Cold Spring Harbor Laboratory) · 2026-05-14

  articleOpen accessCorresponding

  Abstract Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive liver disease for which the mechanisms linking lipid dysregulation to fibrosis remain poorly defined. Hepatic phosphatidylcholine (PC) content is reduced in MASH, but how this alteration drives disease progression is unclear. Here, we identify a role for copper (Cu) homeostasis as a downstream effector of impaired PC biosynthesis. Using single-nucleus RNA sequencing in complementary genetic and dietary mouse models, we found that reduced hepatic PC is associated with marked depletion of hepatic Cu and a concomitant increase in circulating Cu, indicating disrupted Cu distribution. Mechanistically, PC depletion impaired plasma membrane localization of the high-affinity Cu transporter CTR1 ( SLC31A1 ) in hepatocytes, limiting Cu uptake. In human hepatic stellate cells, Cu promoted fibrogenic activation, whereas suppression of Cu import or pharmacologic inhibition of MAPK signaling attenuated fibronectin deposition. In vivo , liver-directed Cu supplementation restored hepatic Cu levels and reduced steatosis but failed to improve fibrosis. In contrast, pharmacologic Cu chelation with bathocuproinedisulfonic acid (BCS) reduced fibrosis without affecting inflammation. Together, these findings identify Cu redistribution as a consequence of impaired PC biosynthesis and implicate Cu-dependent signaling in stellate cell activation, fibrogenesis and MASH pathogenesis. Graphical Abstract

  PublisherDOI

• Mitochondrial copper stabilizes lipoylated TCA cycle proteins to sustain metabolism and proliferation

  bioRxiv (Cold Spring Harbor Laboratory) · 2026-05-22

  articleOpen accessSenior authorCorresponding

  Copper (Cu) is an essential cofactor for mitochondrial cytochrome c oxidase, yet whether it directly regulates mitochondrial metabolism beyond respiration remains unclear. Here we show that mitochondrial Cu, delivered by SLC25A3, is required to maintain the stability of lipoylated TCA cycle proteins. Loss of Slc25a3 or pharmacological Cu depletion selectively destabilized the lipoylated E2 subunits of mitochondrial dehydrogenases and the lipoylation enzymes LIPT1 and LIPT2, an effect not reproduced by acute electron transport chain inhibition. Mechanistically, we find that Cu directly engages the reduced lipoyl moiety using chemical probes and synthetic peptide approaches. Cu depletion impaired PDH and OGDH activity, rewired TCA cycle metabolism, and imposed a dependence on pyruvate carboxylase for anaplerosis. This metabolic defect depleted aspartate, suppressed mTORC1 signaling, and limited proliferation. Conversely, selective delivery of Cu to the mitochondria restored lipoylation, TCA cycle function, and cell growth. Together, these findings identify mitochondrial Cu as a structural regulator of the lipoylation machinery and reveal a direct link between Cu homeostasis and central carbon metabolism.

  PublisherDOI

• Making Every Penny Count: Kinase Signaling Transduction, Copper Homeostasis, & Nutrient Sensing

  Journal of Molecular Biology · 2025-03-13 · 3 citations

  articleOpen access1st authorCorresponding

  • Legacy of Resilience and Representation: Growing up in Virginia, my family's resilience inspired me, but the lack of STEM representation initially limited my awareness of research opportunities.. • Pivotal Research Experience at UNC-CH: As summer research fellowship at UNC-Chapel Hill in Dr. Channing Der’s lab introduced me to biomedical research, igniting my passion for scientific discovery and shaping my decision to pursue a Ph.D. in Pharmacology. • Graduate Research on RAS Signaling and Tumorigenesis: At UNC-CH, I studied the role of Rho GTPases in cancer biology under Dr. Adrienne Cox, discovering a novel mechanism by which an atypical Rho GTPase co-opts epithelial polarity proteins to drive tumorigenesis. • Postdoctoral Training and Discovery of Copper’s Role in Kinase Signaling: My postdoctoral work with Dr. Christopher Counter at Duke University expanded my expertise in oncogenic signaling, leading to the discovery that copper regulates MAPK signaling and influences tumorigenesis. • Commitment to Interdisciplinary Science and Mentorship: Throughout my training, diverse collaborations reinforced the value of interdisciplinary science, while my experiences underscored the importance of mentorship, representation, and fostering inclusive scientific environments. I am the Harrison McCrea Dickson, MD, and Clifford C. Baker, MD Presidential Associate Professor of Cancer Biology at the University of Pennsylvania Perelman School of Medicine. I earned a BS in Chemistry from Radford University and a PhD in Pharmacology from UNC-Chapel Hill before completing postdoctoral training at Duke University with Dr. Christopher Counter. At Penn, I lead a research program pioneering metalloallostery , where redox-active metals regulate kinase activity. We investigate the intersection of kinase signaling and copper (Cu) homeostasis, identifying Cu-dependent kinases and developing targeted therapies through drug repurposing and novel drug design. Our work has advanced our understanding of metals in nutrient signaling, energy homeostasis, and cancer metabolism. I am a Pew Biomedical Scholar, a V Foundation Scholar, and the recipient of the Perelman School of Medicine’s Michael S. Brown New Investigator Research Award. I am also a dedicated advocate for diversity, equity, inclusion, and accessibility (DEIA), having spent the past decade addressing barriers to representation in STEM. In 2021, I was appointed the inaugural Assistant Dean for Inclusion, Diversity, and Equity (IDE) in Research Training at Penn, leading efforts to foster an inclusive research environment. For these contributions, I was recognized with the 2022 Vanderbilt Basic Science Juneteenth Icon Award and the Penn Biomedical Graduate Studies Cell and Molecular Biology Graduate Group Community Service Award.

  PublisherDOI

• Abstract 2535 Mitochondrial matrix Cu transport is required for cytoplasmic signaling and metabolic reprogramming

  Journal of Biological Chemistry · 2025-05-01

  articleOpen accessSenior author

  The transition metal copper (Cu) is an essential catalytic cofactor in the active sites of Cu-dependent enzymes responsible for a diverse range of biological functions including mitochondrial respiration and cellular proliferation. In the case of mitochondria, Cu import is facilitated by the dual Cu and phosphate carrier, SLC25A3, whose knockout causes loss in activity of Cu-dependent enzymes like the cytochrome c oxidase (COX) complex. Given that Cu homeostasis is integral to normal cell survival as evidenced by Menkes disease, surprisingly little is known about Cu-sensing mechanisms and their interaction with key signaling molecules that translates copper abundance to distinct cellular functions.

  PublisherOA PDFDOI

• ER stress tolerance is regulated by copper-dependent PERK kinase activity

  Cell Reports · 2025-09-27 · 2 citations

  articleOpen access

  Pancreatic/PKR-like endoplasmic reticulum (ER) kinase (PERK) is a kinase that, in response to ER stress, mediates dual homeostatic and pro-apoptotic signaling. Thus, intricate regulation is required for physiological function. Attempts to modulate PERK activity have shown that the determinants of adaptive vs. maladaptive signaling remain ambiguous. Here, with purified protein, we provide evidence that PERK binds copper, identifies residues required for interaction, and demonstrates that copper is necessary for kinase activity. Furthermore, cellular PERK activity can be modulated via copper availability, and this regulatory relationship can be manipulated to dictate ER stress tolerance. Critically, these phenomena translate to phenotypes in vivo, as C. elegans harboring a "PERK-copper mutant" exhibit exacerbated ER-stress sensitivity. The copper-PERK paradigm suggests that copper homeostasis, as a regulator of PERK, may constitute a critical factor in resolving the long-standing ambiguity in endeavors to therapeutically target PERK.

  PublisherDOI

• Juneteenth in STEMM and the barriers to equitable science

  UNC Libraries · 2025-03-19

  articleOpen access1st authorCorresponding
  PublisherDOI

• A histochemical approach to activity-based copper sensing reveals cuproplasia-dependent vulnerabilities in cancer

  Proceedings of the National Academy of Sciences · 2025-01-15 · 14 citations

  articleOpen access

  Copper is an essential nutrient for sustaining vital cellular processes spanning respiration, metabolism, and proliferation. However, loss of copper homeostasis, particularly misregulation of loosely bound copper ions which are defined as the labile copper pool, occurs in major diseases such as cancer, where tumor growth and metastasis have a heightened requirement for this metal. To help decipher the role of copper in the etiology of cancer, we report a histochemical activity-based sensing approach that enables systematic, high-throughput profiling of labile copper status across many cell lines in parallel. Coppermycin-1 reacts selectively with Cu(I) to release puromycin, which is then incorporated into nascent peptides during protein translation, thus leaving a permanent and dose-dependent marker for labile copper that can be visualized with standard immunofluorescence assays. We showcase the utility of this platform for screening labile Cu(I) pools across the National Cancer Institute's 60 (NCI-60) human tumor cell line panel, identifying cell types with elevated basal levels of labile copper. Moreover, we use Coppermycin-1 to show that lung cancer cells with heightened activation of nuclear factor-erythroid 2-related factor 2 (NRF2) possess lower resting labile Cu(I) levels and, as a result, have reduced viability when treated with a copper chelator. This work establishes that methods for labile copper detection can be used to assess cuproplasia, an emerging form of copper-dependent cell growth and proliferation, providing a starting point for broader investigations into the roles of transition metal signaling in biology and medicine.

  PublisherDOI

• MAPK Pathway Inhibition Reshapes Kinase Chemical Probe Reactivity Reflecting Cellular Activation States

  bioRxiv (Cold Spring Harbor Laboratory) · 2025-10-23

  preprintOpen accessSenior authorCorresponding

  Despite the pivotal role of oncogenic kinases in cancer initiation, progression, and therapeutic resistance, functionally profiling their activity and conformational dynamics in live cells remains challenging. Existing methods often fail to capture inhibitor-bound structural states of kinases, particularly in clinically relevant contexts, such as treatment response and acquired resistance, where genomic data alone are insufficient. Here, we use activity-based protein profiling (ABPP) to monitor composite amino acid reactivity changes, across cysteine, lysine, and carboxylic acid residues, as a hypothesis-generating readout of kinase state in live cells. Using electrophilic probes, we show that treatment of BRAFV600E mutant melanoma cells with vemurafenib or trametinib decreases overall cysteine and lysine reactivity in BRAFV600E and MEK1/2, likely reflecting composite changes in amino acid accessibility across multiple reactive residues associated with inhibitor binding. Changing the order of probe addition and inhibitor treatment altered the labeling outcomes, consistent with competitive engagement and structural stabilization. Comparative analysis of ATP-competitive BRAFV600E inhibitors vemurafenib and dabrafenib indicated differences in aspartate and glutamate labeling patterns, consistent with the possibility that ABPP may detect inhibitor-associated variations in residue accessibility, which could reflect differences in inhibitor-bound conformations. In inhibitor-resistant melanoma models, ABPP detected differences in residue reactivity relative to parental cells, which aligned with known resistance-associated features, such as BRAF overexpression and the MEK2 Q60P activation mutation. Moreover, global proteome analyses of cysteine and lysine reactivity upon BRAFV600E inhibition revealed probe-accessible cysteine labeling changes on KSR2, suggesting a potential MAPK pathway remodeling. Together, these findings highlight ABPP as a valuable chemical biology approach for investigating inhibitor-dependent changes in kinase residue reactivity, offering a framework to investigate how kinase conformational dynamics and signaling pathway adaptation influence the therapeutic response and resistance in cancer.

  PublisherDOI

• A selective S-acyltransferase inhibitor suppresses tumor growth

  bioRxiv (Cold Spring Harbor Laboratory) · 2024-07-22 · 2 citations

  preprintOpen access

  S-acyltransferases play integral roles in essential physiological processes including regulation of oncogenic signaling pathways. While discovered over 40 years ago the field still lacks specific S-acylation inhibitors thus the potential benefit of pharmacologically targeting S-acyltransferases for human disease is still unknown. Here we report the identification of an orally bioavailable acyltransferase inhibitor SD-066-4 that inhibits the acyltransferase ZDHHC20. We identified a specific alanine residue that accommodates the methyl group of SD-066-4, thus providing isoform selectivity. SD-066-4 stably reduces EGFR S-acylation in Kras mutant cells and blocks the growth of Kras mutant lung tumors extending overall survival. We find that lung cancer patients harboring deletions in ZDHHC20 or ZDHHC14 concurrent with Kras alterations have a significant survival benefit, underscoring the translational importance of these enzymes.

  PublisherOA PDFDOI

• Figure S9 from Inhibition of BCL2 Family Members Increases the Efficacy of Copper Chelation in BRAF<sup>V600E</sup>-Driven Melanoma

  2023-12-22

  preprintOpen accessSenior author

  <p>Supplemental Figure S9. Cotreatment with TTM and ABT-263 suppresses in vivo growth of parental and MAPK inhibitor resistant BRAFV600E-positive melanoma.</p>

  PublisherOA PDFDOI

Recent grants

• NIH Grant K01CA178145

  NIH · $274k · 2016

• Molecular and Cellular Mechanisms of Copper-Dependent Nutrient Signaling and Metabolism

  NIH · $4.1M · 2017–2027

Frequent coauthors

• Tiffany Tsang

  63 shared

• Jessica M. Posimo

  58 shared

• Ye‐Jin Kim

  30 shared

• Caroline I. Davis

  29 shared

• Christopher M. Counter

  Duke Medical Center

  27 shared

• Ronen Marmorstein

  22 shared

• Xingxing Gu

  Washington University in St. Louis

  20 shared

• Grace R. Anderson

  Texas A&M University

  20 shared

Labs

• Brady LabPI