About

Giulia Pavani, PhD, is a Research Assistant Professor of Pathology and Laboratory Medicine at the University of Pennsylvania's Perelman School of Medicine. She completed her undergraduate studies with a BA in Cell and Molecular Biology from the University of Ferrara in Italy in 2008, followed by a BS in Molecular and Cellular Biotechnologies from University Vita-Salute S.Raffaele in Milan in 2010. She earned her PhD in Molecular Biology and Biotechnology from the University of Ferrara in 2014. Her research expertise includes induced pluripotent stem cells, hematopoiesis, and gene editing. Dr. Pavani's work focuses on modeling erythropoiesis with induced pluripotent stem cells, producing red blood cells from pluripotent stem cells for precision transfusion medicine, and exploring the molecular mechanisms underlying hematopoietic stem cell development. She has contributed to understanding cell state transitions during hematopoiesis and the signaling pathways involved in hematopoietic stem and progenitor cell development. Her research aims to advance regenerative medicine and transfusion medicine through innovative stem cell and gene editing technologies.

Research topics

• Medicine
• Internal medicine
• Cell biology
• Biology
• Cancer research
• Genetics
• Immunology

Selected publications

• Rare antigen‐negative red blood cells from pluripotent stem cells for precision transfusion medicine

  Transfusion · 2026-04-24

  articleOpen access

  BACKGROUND: Blood bank identification of antibodies against high-prevalence antigens remains a challenge due to the scarcity of antigen-negative reagent red cells sourced from blood donors. The MAM antigen, encoded by EMP3, is one such antigen associated with red cell alloimmunization and hemolytic disease of the fetus and newborn. STUDY DESIGN AND METHODS: We used CRISPR-Cas9 gene editing to generate an EMP3 knockout (EMP3KO) induced pluripotent stem cell (iPSC) line from a type O, Rh null parent line, enabling production of rare MAM-negative red blood cells. Since a prior study suggested that loss of EMP3 may enhance erythroid proliferation, we hypothesized that EMP3KO could both yield a rare reagent cell and potentially improve erythroid expansion to support scalable production. Transcriptomic analysis allowed us to further investigate the effect of EMP3 loss in late erythroblasts. RESULTS: EMP3KO cells differentiated efficiently into erythroid cells, showing >95% CD235/CD71 co-expression and orthochromatic erythroblast morphology. Compared to unedited cells, no proliferative advantage was observed, contrasting with prior non-isogenic cell models. Agglutination assays confirmed complete loss of MAM antigen and demonstrated the diagnostic utility for identifying MAM antibodies. Transcriptomic profiling of EMP3KO erythroblasts revealed expression of key erythroid genes, as well as regulators of proliferation and heme metabolism, was comparable to the parent line. DISCUSSION: This study demonstrates that iPSC technology combined with gene editing can generate rare antigen-negative RBCs for immunohematology applications. Beyond MAM, this platform offers a strategy to create additional rare RBC phenotypes, advancing precision transfusion medicine and improving antibody identification against high-prevalence antigens.

  PublisherDOI

• Novel cytometry-based characterization of lysosomal storage disease affected patient's cells

  bioRxiv (Cold Spring Harbor Laboratory) · 2025-03-18

  preprintOpen access

  Wolman disease (WD) is a severe lysosomal storage disorder characterized by fatal lipid accumulation caused by the deficiency of a lipid metabolic enzyme, Lysosomal Acid Lipase (LAL), involved in the lysosomal hydrolysis of cholesterols and triglycerides. Due to the imbalance of lipids homeostasis, WD patients suffer from severe hepatosplenomegaly, hepatic failure and adrenal calcification resulting in a premature infant death within the first year of age. In this work, we explored multiple imaging analyses to fully characterize the phenotype of LAL deficient cells. In particular, we stained WD patients' fibroblasts for intracellular lipid droplets (LD) and lysosomes and we analysed staining intensity and granularity as well as an increased number of LD and lysosomes using fluorescence wide field microscopy, confocal microscopy, conventional and image flow cytometry. Noteworthy, we showed that lipid homeostasis was restored upon delivery of a functional LAL transgene. Finally, since fibroblasts cannot be used as routine clinical test as they are difficult to collect from WD patients, we confirmed our observations in LAL deficient human blood cell lines and in peripheral blood mononuclear cells (PBMC) from LAL deficient (LAL-D) mouse model, as a proxy for easily accessible WD PBMC. Overall, we expect that this novel imaging analysis pipeline will help to diagnose WD, follow its progression and evaluate the success of enzyme replacement therapy or gene correction strategies for WD as well as other lysosomal storage disorders.

  PublisherOA PDFDOI

• Using iPSC-derived hematopoietic stem cells with long-term engraftment capability to model fetal blood disorders

  Blood · 2025-11-03

  articleOpen access1st authorCorresponding

  Abstract Introduction: Human developmental hematopoiesis is a complex process occurring in sequential waves at different embryonic sites. This process yields both differentiated blood cells essential for embryonic development and hematopoietic stem cells (HSCs) necessary for lifelong blood cell production. There are at least two main hematopoietic programs during embryogenesis: a transient primitive wave that primarily generates myelo-erythroid progenitors in the yolk sac, and a definitive wave occurring in the aorta-gonad-mesonephros region which produces progenitors with expanded lineage potential and the first transplantable HSCs. A challenge in generating HSCs from induced pluripotent stem cells (iPSCs) is that many current methods recapitulate early developmental stages only, failing to produce transplantable HSCs and limiting their application for in vivo studies. We have previously shown that we can recapitulate fetal erythropoiesis using a protocol that produces definitive hematopoietic cells. Red cells generated with this method primarily produce fetal globin and are functionally distinct from primitive erythroblasts. Here, we adapted this method to a serum-free 3D culture system to produce definitive hematopoietic stem progenitor cells (HSPCs) from iPSCs to 1) evaluate long-term engraftment via xenotransplantation and 2) model a preleukemic disorder of fetal origin, transient abnormal myelopoiesis (TAM), which affects ~20% of neonates with Trisomy 21 (T21) and is associated with mutations in the key hematopoietic transcription factor GATA1. Methods We designed a serum-free 3D culture system that directs mesodermal commitment through Wnt pathway activation by manipulating developmental cues through retinoic acid signaling and shear stress. We transplanted 24 NBSGW mice with 1.2-2 million iPSC-derived HSPCs using 3 distinct wildtype (WT) lines and experimental batches. The 3D definitive differentiation was assessed using isogenic T21 iPSCs with wild type GATA1 or the truncated isoform lacking the N-terminus, GATA1s. Results Long-term repopulation was achieved up to 20 weeks post-transplant in the bone marrow (BM) of all engrafted mice (mean = 3.9±11.7% human HLA-ABC+) and in the peripheral blood of 16/24 mice. In mice with the highest level of engraftment (>10%), we detected human HSCs as well as myeloid, lymphoid, erythroid and megakaryocyte precursors in the BM, indicating complete hematopoietic reconstitution. Human CD34+ cells recovered from the graft retained multilineage potential in colony forming assays. Importantly, no leukemia or teratomas were observed in any of the transplanted mice. To assess whether this definitive iPSC culture protocol could faithfully model the fetal blood disorder TAM, we generated definitive HSPCs from T21/wtGATA1 and T21/GATA1s iPSCs. Both T21 lines generated budding HSPCs at day 15; however, the percentage of CD34+CD45+ HSPCs was lower compared to euploid controls (60.5% of WT ±12.9 for T21, and 64.5% of WT ± 0.06 for T21/GATA1s). In addition, at day 15 we observed a significant CD41+CD42b+ megakaryocyte population in both trisomy lines, consistent with the disease phenotype and that was not previously observed with our primitive hematopoietic differentiation protocols. Compared to WT controls, we observed a 2.3-fold and a 4.2-fold increased megakaryocyte population from T21/wtGATA1 and T21/GATA1s iPSCs, respectively. T21/GATA1s HPSCs also showed aberrant morphology with megakaryocytic and blast-like features. Conclusions These studies show that our serum-free 3D culture system can generate iPSC-derived HSPCs that can engraft immunodeficient mice, reconstitute different blood lineages, and persist for 20 weeks. Using a T21/TAM iPSC model, we found an enhanced megakaryocyte population in T21/wtGATA1 consistent with a phenotype we observed with primary human fetal liver hematopoiesis, but not with iPSC-derived primitive hematopoietic progenitors. Thus, T21/GATA1s definitive HSPCs recapitulate the enhanced megakaryopoiesis consistent with TAM in vitro. Ongoing xenotransplant experiments will help elucidate the interaction of trisomy 21 and GATA1s and provide a novel in vivo model to study and treat human hematological diseases.

  PublisherOA PDFDOI

• Generation of CHOPi014-A from healthy adult peripheral blood mononuclear cells

  Stem Cell Research · 2025-08-24

  articleOpen accessSenior authorCorresponding

  CHOPWT15is a control male induced pluripotent stem cell (iPSC) line that can be used to model genetic variants by creating an allelic series of isogenic lines using genome editing technologies (Hockemeyer and Jaenisch, 2016; Maguire et al., 2022; Pavani et al., 2022). An allelic series of iPSC lines provides the means to perform genotype and phenotype analyses of pathological variants without the confounding effects of genetic background. Peripheral blood mononuclear cells (PBMCs), obtained from a healthy adult male, were reprogrammed using Sendai virus creating an iPSC line that has undetectable copy number variations (CNVs, resolution >= 100 kb) and differentiates to all three germlayers.

  PublisherDOI

• Advanced Imaging and Cytometric Techniques to Characterize Lipid Accumulation in Wolman Disease

  Cytometry Part A · 2025-07-01

  articleOpen access

  Wolman disease (WD) is a severe lysosomal storage disorder characterized by fatal lipid accumulation caused by the deficiency of a lipid metabolic enzyme, Lysosomal Acid Lipase (LAL), involved in the lysosomal hydrolysis of cholesterols and triglycerides. Due to the imbalance of lipid homeostasis, WD patients suffer from severe hepatosplenomegaly, hepatic failure, and adrenal calcification resulting in a premature infant death within the first year of age. In this work, we explored multiple imaging analyses to fully characterize the phenotype of LAL-deficient cells. In particular, we stained WD patients' fibroblasts for intracellular lipid droplets (LD) and lysosomes, and we analyzed staining intensity and granularity, as well as an increased number of LD and lysosomes using fluorescence wide-field microscopy, confocal microscopy, conventional, and image flow cytometry. Noteworthy, we showed that lipid homeostasis was restored upon delivery of a functional LAL transgene. Finally, since fibroblasts cannot be used as routine clinical tests as they are difficult to collect from WD patients, we confirmed our observations in LAL deficient human blood cell lines and in peripheral blood mononuclear cells (PBMC) from the LAL deficient (LAL-D) mouse model, as a proxy for easily accessible WD PBMC. Overall, we expect that this novel imaging analysis pipeline will help to diagnose WD, follow its progression, and evaluate the success of enzyme replacement therapy or gene correction strategies for WD as well as other lysosomal storage disorders.

  PublisherOA PDFDOI

• Sanctions, tariffs, and trade wars: The role of geopolitical tensions in global business dynamics

  International Journal of Research in Management · 2025-01-01 · 1 citations

  articleOpen accessSenior author

  Global business operations now experience increased influence from geopolitical conflicts because of sanctions alongside tariff barriers and trade warfare. Economic instruments that governments put to use frequently as strategic tools modify the direction of international trade while influencing both investment patterns and corporate choices. Trade sanctions also make it difficult to do international deals. For particular regions, this disrupts domestic businesses and disrupts manufacturing pipelines. It not only destroys some businesses but also promotes other benefits. Barriers on importing into our country suggested by the administration to defend our domestic industry will result in similar barriers to trade in other countries. Threats to foreign businesses in their respective home markets will lead to countermeasures which will eventually destabilize global business markets. The United States’ fight with China on trade, which has just been heating up over the past week, has led to growing economic uncertainties. The currencies of nations dwindled, their stock markets waned, and business confidence fell. Particular industrial sectors have been advantaged by protectionists during this period put multitudes of business sectors at a disadvantage by making it difficult to operate or more costly to operate, and impeded access to valuable resources. Corporate entities that operate across boundaries flow to this shifting landscape which necessitates the rapid shifting of rules and the reorganization of distribution networks.

  PublisherOA PDFDOI

• Synergistic cooperation between progranulin and Jak2/Stat3 signaling determines definitive myeloid cell fate

  Blood · 2025-11-03

  article

  Abstract Perturbations in cell fate choices result in loss of tissue homeostasis and diseases like cancer and developmental disorders. A better comprehension of the molecular mechanisms that drive cell fate determination is necessary to reveal novel therapeutic interventions. The JAK/STAT signaling pathway is a cornerstone to cancer progression, including hematopoietic malignancies. However, little is known about the molecular regulators of this pathway, and how they enable hematopoietic cell state transitions in homeostatic conditions and disease. In this study, we found in vivo that progranulin (GRN), a gene associated to human frontotemporal dementia, is critical to determine myeloid cell fate through the regulation of the JAK2/STAT3 pathway. Through the generation of inducible stat3 and progranulin a (grna) zebrafish, in conjunction with knockout ablation models, FACS sorting, transcriptomic profiling, rescue experiments, and CUT&RUN, we demonstrate in vivo that Stat3 induces grna expression,which in turns increases stat3 expression and reinforces myeloid fate commitment over erythroid differentiation. In addition, in vivo lineage tracing experiments coupled with live imaging and tissue-specific ablations show that the Grna/Stat3 axis operates during definitive, but not primitive myelopoiesis. This distinct molecular requirement during developmental myelopoiesis helped identify key functional differences between macrophages from different embryonic origins. Particularly, we found that definitive myeloid cells are the main contributors of tissue repair and organogenesis, while primitive myeloid cells are functionally inefficient in these contexts. Finally, successful myeloid differentiation of a human myelogenous leukemia line was only achieved when the JAK2/STAT3 pathway was co-stimulated with GRN, showing high conservation during human myelopoiesis and the critical role of GRN during myeloid fate determination. This discovery reveals a new mechanism to control hematopoietic lineage commitment in homeostatic conditions, and uncovers progranulin as a potential target to correct hematological malignancies with faulty IL-6/JAK2/STAT3 activation.

  PublisherDOI

• CRISPR-mediated transcriptional activation as a mutation-independent therapeutic strategy for <i>SYNGAP1</i> -related intellectual disability

  bioRxiv (Cold Spring Harbor Laboratory) · 2025-10-29 · 2 citations

  preprintOpen access

  Abstract Synaptic Ras GTPase-activating protein (SynGAP) regulates synaptic strength and neuronal signaling, with essential roles in cortical development and synaptic plasticity. Heterozygous loss-of-function variants in SYNGAP1 cause SYNGAP1 -related intellectual disability (SRID), a severe neurodevelopmental disorder characterized by epilepsy, developmental delay, and autism. SYNGAP1 mutations often result in haploinsufficiency, providing a strong rationale for gene-targeted therapies. However, no treatment currently addresses the underlying genetic cause of SRID. Here, we developed a CRISPR-mediated transcriptional activation (CRISPRa) approach to upregulate the functional Syngap1 allele in a SRID mouse model. CRISPRa activated Syngap1 , normalized SynGAP protein expression and downstream signaling, and rescued working memory deficits. We validated the translational potential of this strategy in human induced pluripotent stem cell (hiPSC)-derived excitatory cortical neurons. CRISPRa rescued SYNGAP1 in two distinct loss-of-function variant lines. Together, these findings demonstrate the feasibility of mutation-independent transcriptional activation as a therapeutic approach for SRID and its broader applicability to haploinsufficiency disorders.

  PublisherDOI

• 3048 – NOD1-DEPENDENT NF-KB ACTIVATION INITIATES HEMATOPOIETIC STEM CELL SPECIFICATION IN RESPONSE TO SMALL RHO GTPASES

  Experimental Hematology · 2024-08-01

  article
  PublisherDOI

• Modeling primitive and definitive erythropoiesis with induced pluripotent stem cells

  Blood Advances · 2024-01-30 · 15 citations

  articleOpen access1st authorCorresponding

  ABSTRACT: During development, erythroid cells are produced through at least 2 distinct hematopoietic waves (primitive and definitive), generating erythroblasts with different functional characteristics. Human induced pluripotent stem cells (iPSCs) can be used as a model platform to study the development of red blood cells (RBCs) with many of the differentiation protocols after the primitive wave of hematopoiesis. Recent advances have established that definitive hematopoietic progenitors can be generated from iPSCs, creating a unique situation for comparing primitive and definitive erythrocytes derived from cell sources of identical genetic background. We generated iPSCs from healthy fetal liver (FL) cells and produced isogenic primitive or definitive RBCs which were compared directly to the FL-derived RBCs. Functional assays confirmed differences between the 2 programs, with primitive RBCs showing a reduced proliferation potential, larger cell size, lack of Duffy RBC antigen expression, and higher expression of embryonic globins. Transcriptome profiling by scRNA-seq demonstrated high similarity between FL- and iPSC-derived definitive RBCs along with very different gene expression and regulatory network patterns for primitive RBCs. In addition, iPSC lines harboring a known pathogenic mutation in the erythroid master regulator KLF1 demonstrated phenotypic changes specific to definitive RBCs. Our studies provide new insights into differences between primitive and definitive erythropoiesis and highlight the importance of ontology when using iPSCs to model genetic hematologic diseases. Beyond disease modeling, the similarity between FL- and iPSC-derived definitive RBCs expands potential applications of definitive RBCs for diagnostic and transfusion products.

  PublisherOA PDFDOI

Frequent coauthors

• Annarita Miccio

  Hôpital Necker-Enfants Malades

  31 shared

• Deborah L. French

  19 shared

• Paris Margaritis

  University of Pennsylvania

  18 shared

• Paul Gadue

  Children's Hospital of Philadelphia

  16 shared

• Mario Amendola

  Inserm

  15 shared

• Chiara Antoniani

  Institut des Maladies Génétiques Imagine

  14 shared

• Vasco Meneghini

  Vita-Salute San Raffaele University

  13 shared

• Tristan Félix

  Institut des Maladies Génétiques Imagine

  12 shared

Labs

• Pathology and Laboratory MedicinePI