Ex-vivo Stem Cell Expansion and Directed Differentiation

Founded in December 2015 with licensed technology developed and studied for over two decades at Fred Hutchinson Cancer Research Center, Nohla’s proprietary technology is based on the density-dependent Notch ligand platform that uses purified CD34+ cord blood stem and progenitor cells cultured in the presence of an engineered form of the Notch ligand Delta1 (Delta1ext-IgG or DXI).

This platform enables ex vivo expansion, directed stem cell fate and lineage-specific differentiation of hematopoietic stem and progenitor cells with broad application across multiple cell types and therapeutic areas.

Hematopoietic Stem/Progenitor Cell Expansion

The role of Notch in Hematopoiesis

In the early 1990’s Dr. Irwin Bernstein at the Fred Hutchinson Cancer Research Center postulated that, as in other developing systems, hematopoietic stem cell-fate specification resulted from intercellular interactions with adjacent cells and was modulated by several families of molecules, including the Notch gene family. At that time, the Notch pathway was particularly well-studied in invertebrate systems with clear evidence that Notch played an important role in mediating intercellular interactions affecting cell-fate decisions within the central nervous system, eye, mesoderm, and ovaries.

A role for Notch in hematopoiesis was then further suggested by Dr. Bernstein’s detection of the human Notch1 gene in CD34+ or CD34+lin− human hematopoietic precursors. Subsequently, primary murine hematopoietic stem cells (HSC) were transduced with a retrovirus leading to constitutively active expression of the intracellular domain (ICD) of Notch1 and led to the emergence of an immortalized pluripotent cytokine-dependent cell line capable of both lymphoid and myeloid repopulation in vivo, thereby demonstrating a role for Notch in hematopoietic stem progenitor cell (HSPC) self-renewal. Although at the time a biological function for Notch in HSPC was not determined, this work suggested that manipulation of the Notch signaling pathway ex vivo in primary HSC could prove to be a novel approach for expanding HSPC.

Delta1 Ligand

To avoid the potential safety concerns of retroviral transduction, it was decided to activate endogenous Notch signaling in primary HSPC during culture. To that end, engineered Notch ligands were developed consisting of the extracellular domain (ECD) of the Notch ligands Jagged1 and Delta1. Notch ligands activate their receptors by physically pulling on the ECD in vivo; thus immobilization of the ligand proved necessary to activate endogenous Notch signaling in vitro. Using an immobilized form of the Notch ligand Delta1, Dr. Bernstein’s group was able to sufficiently activate endogenous receptors and induce expansion of murine stem progenitor cells capable of in vivo reconstitution similar to that seen in the retroviral-mediated Notch overexpression experiments. In fact, culture of murine hematopoietic precursors with the immobilized ligand Delta1 and cytokines resulted in a several-log increase in progenitors capable of short-term lymphoid and myeloid repopulation.

Cord Blood Expansion

Given the clinical need for large numbers of hematopoietic progenitor cells capable of providing rapid myeloid recovery in vivo, especially in patients undergoing cord blood transplant (CBT), Nohla Scientific Founder and Chief Medical Officer Dr. Colleen Delaney extended the ex vivo expansion platform from murine to human HSPC where she noted a unique response of cord blood (CB) HSPC to Notch ligands as compared to CD34+ bone marrow cells or mobilized peripheral blood stem cells. She found that culture of isolated CB CD34+ HSPC in the presence of an engineered form of Delta1 ligand in serum free media supplemented with cytokines led to a greater than 2-log increase in the number of CD34+ cells and nearly a 16-fold increase in NOD/SCID mouse repopulating cell frequency compared to control.

The ability of these human cells to rapidly reconstitute (as early as 10 days post infusion) the myeloid compartment in immunodeficient mice indicated their potential clinical utility. In contrast to the studies using primary murine HSPCs, in vivo persistence of transplanted cells at 9 weeks and secondary transplantation studies suggested the presence of both long-term and short-term repopulating cells following culture of human CB progenitor cells on Delta ligand.

A key aspect of these studies was determination of whether the magnitude of Notch signaling played a role in the optimal generation of repopulating cells. Murine studies showed that the relatively lower amount of Notch signaling induced in cells cultured with lower densities of Delta-1 led to self-renewal of progenitors with primarily B-lymphoid and myeloid potential, whereas higher amounts of Notch signaling inhibited B cell differentiation and promoted differentiation towards the T cell lineage.

Concurrent studies performed with human HSPC also revealed important ligand dose-dependent effects whereby relatively lower densities of immobilized ligand substantially enhanced generation of NOD/SCID repopulating cells, with higher ligand densities promoting differentiation towards the T-cell lineage at the expense of repopulating cells.

Scientific Publications

Milano F, Gooley T, Wood B, Woolfrey A, Flowers ME, Doney K, Witherspoon R, Mielcarek M, Deeg JH, Sorror M, Dahlberg A, Sandmaier BM,Salit R, Petersdorf E, Appelbaum FR, Delaney CCord-Blood Transplantation in Patients with Minimal Residual DiseaseN Engl J Med. 2016 Sep 8;375(10):944-53.

Oliver DCK, Cassaday RD, Ermoian RP, Dahlberg A, Delaney C, Milano F. Myeloablative Cord Blood Transplantation Yields Excellent Disease Free Survival in Patients with Acute Lymphoblastic Leukemia. Blood 2016; 128:4693

Cox E, Qian VW, Salit R, DelaneyC, Milano F. Infusion of Ex-Vivo Expanded Cord Blood Progenitor Cells Reduces the Time to Platelet Engraftment and the Number of Platelet and Red Blood Cell Transfusions Required in Patients Receiving Myleloblative Cord Blood Transplants. Blood 2016; 128:1232

Merriam F, Imren S, Landeros R, Delaney C. Demonstration of Rapid Functional Myeloid Cell Recovery after Infusion of a Universal Donor Ex Vivo Expanded Progenitor Cell Therapy as a Bridging Graft Source. Blood 2016; 128:3348

Delaney C, Milano F, Cicconi L, Othus M, Becker PS, Sandhu V, Nicoud I, Dahlberg A, Bernstein IDB, Appelbaum FR, Estey EH. Infusion of a non-HLA-matched ex-vivo expanded cord blood progenitor cell product after intensive acute myeloid leukaemia chemotherapy: a phase 1 trialLancet Haematol. 2016 Jul;3(7):e330-9. Epub 2016 Jun 7.

Milano F, Heimfeld S, Riffkin I, Nicoud I, Appelbaum FR, , Bernstein ID, Delaney C. Infusion of a Non HLA-Matched Off-the-Shelf Ex Vivo Expanded Cord Blood Progenitor Cell Product following Myeloablative Cord Blood Transplantation is Safe, Decreases the Time to Hematopoietic Recovery, and Results in Excellent Overall Survival. Blood 2014; 124:46.

Dahlberg A, Milano F, Delaney C. Infusion of ex vivo expanded cord blood progenitor cells is associated with reduced hospital days and utilization of opiate infusion and total parental nutrition in pediatric patients receiving myeloablative cord blood transplantation. Biol Blood Marrow Transplant 2015; Vol. 21, Issue 2, S94–S95.

Delaney C, Heimfeld S, Brashem-Stein C, Voorhies H, Manger R and Bernstein ID. Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nature Medicine, 2010 Feb;16(2):232-6.

Milano, F., Merriam, F., Nicoud, I., Li, J., Gooley, T. A., Heimfeld, S., Imren, S., Delaney, C.  Notch-Expanded Murine Hematopoietic Stem and Progenitor Cells Mitigate Death from Lethal Radiation and Convey Immune Tolerance in Mismatched Recipients Stem Cells Transl Med. sctm.2016-0112

Dahlberg A, Woo S, Delaney C, Boyle P, Gnirke A, Bock C, Bernstein BE, Meissner A, Gottardo R, Bernstein ID. Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity.

Delaney C, Milano F, Cicconi L, Othus M, Becker PS, Sandhu V, Nicoud I, Dahlberg A, Bernstein IDB, Appelbaum FR, Estey EH. Infusion of a non-HLA-matched ex-vivo expanded cord blood progenitor cell product after intensive acute myeloid leukaemia chemotherapy: a phase 1 trial. Lancet Haematol. 2016 Jul;3(7):e330-9. Epub 2016 Jun 7

Milano F, Gooley T, Wood B, Woolfrey A, Flowers ME, Doney K, Witherspoon R, Mielcarek M, Deeg JH, Sorror M, Dahlberg A, Sandmaier BM,Salit R, Petersdorf E, Appelbaum FR, Delaney C. Cord-Blood Transplantation in Patients with Minimal Residual Disease. N Engl J Med. 2016 Sep 8;375(10):944-53.

Oliver DCK, Cassaday RD, Ermoian RP, Dahlberg A, Delaney C, Milano F. Myeloablative Cord Blood Transplantation Yields Excellent Disease Free Survival in Patients with Acute Lymphoblastic Leukemia. Blood 2016; 128:4693

Cox E, Qian VW, Salit R, DelaneyC, Milano F. Infusion of Ex-Vivo Expanded Cord Blood Progenitor Cells Reduces the Time to Platelet Engraftment and the Number of Platelet and Red Blood Cell Transfusions Required in Patients Receiving Myleloblative Cord Blood Transplants. Blood 2016; 128:1232

Merriam F, Imren S, Landeros R, Delaney C. Demonstration of Rapid Functional Myeloid Cell Recovery after Infusion of a Universal Donor Ex Vivo Expanded Progenitor Cell Therapy as a Bridging Graft Source. Blood 2016; 128:3348

Nemecek ER, Adams AJ, Shaw BE, Kiefer DM, Le-Rademacher JG, Levine JE, Yanik GA, Leung WH, Talano J-AM, Haut PR, Delgado DC, Kapoor N, Petrovic A, Adams RH, Hanna R, Rangarajan HG, Dalal JD, Chewning JH, Verneris MR, Epstein SS, Pulsipher MA, Delaney C. Phase II Study of Treosulfan/Fludarabine/ Low Dose Total Body Irradiation As a Preparative Regimen for Children with Acute Myeloid Leukemia (AML) or Myelodysplastic Syndrome (MDS) Undergoing Allogeneic Hematopoietic Cell Transplantation. Biol Blood Marrow Transplant 2016; Vol.22, Issue 3, S99

Salit RB, Emerson R, Karanes C, Gutman JA, Nikiforow S, Duncan C, Kurtzberg J, Robins H, Rieder M, Delaney C. Interim Analysis of an Observational Study Assessing T Cell Receptor Diversity As an Early Predictor of NRM in Cord Blood Transplant Recipients. Biol Blood Marrow Transplant 2016; Vol.22, Issue 3, S44

Albert P, Delaney C, Salit R, Anderson N, Milano F. Better High Resolution HLA-Matching Is Associated with Lower Transplant Related Mortality after Cord Blood Transplantation. Blood 2015; 4405.

Oliver DCK, Milano F, Gammill H, Gentil C, Kanaan SB, Allen J, Nelson L, Delaney C. Persistence of the Losing Unit Following Double Umbilical Cord Blood Transplant: Finding the Unseen. Blood 2015; 126:3109.

Hill JA, Mayer BT, Xie H, Leisenring WM, Milano F, Delaney C, Huang M-L, Stevens-Ayers TL, Jerome KR, Nichols G, Zerr DM, Schiffer JT, Boeckh M. Detection of Multiple Double-Stranded DNA Viruses after Cord Blood Transplantation Is Frequent and Persistent.  Blood 2015; 126:3104

Li J, Jiang A, Milano F, Imren S, Oliver D, Blake J, Delaney C. Distinct Patterns of Early Hematopoietic Reconstitution in Patients Receiving Notch Expanded Non HLA-matched Off-The-Shelf Cord Blood Derived Hematopoietic Stem and Progenitor Cells. Blood 2015; 126:379.

Li J, Blake J, Delaney C. Identification of CD137+ IFN-g- CD4+ Alloreactive T Cells Post Myeloablative Double Cord Blood Transplant: A Synergistic Role in the Graft Versus Graft Interaction. Biol Blood Marrow Transplant 2015; Vol.21, Issue 2, S57-S58

Dahlberg A, Milano F, Delaney C. Infusion of ex vivo expanded cord blood progenitor cells is associated with reduced hospital days and utilization of opiate infusion and total parental nutrition in pediatric patients receiving myeloablative cord blood transplantation. Biol Blood Marrow Transplant 2015; Vol. 21, Issue 2, S94–S95.

Salit R, Milano F, Delaney C. Outcomes of Cord Blood Transplant as Salvage Therapy following Graft Failure or Relapse After Prior Allogeneic Transplant.Biol Blood Marrow Transplant. 2016 Feb;22(2):339-43. Epub 2015 Oct 17.2015.

Milano F, Heimfeld S, Riffkin I, Nicoud I, Appelbaum FR, , Bernstein ID, Delaney C. Infusion of a Non HLA-Matched Off-the-Shelf Ex Vivo Expanded Cord Blood Progenitor Cell Product following Myeloablative Cord Blood Transplantation is Safe, Decreases the Time to Hematopoietic Recovery, and Results in Excellent Overall Survival. Blood 2014; 124:46.Li J, Milano F, Blake JM, Oliver DC, Nicoud I, Heimfeld S, Delaney C. Real-Time Immunophenotyping of Peripheral Blood Early Post Myeloablative Cord Blood Transplant Can Identify Patients at Risk for Primary Graft Failure. Blood 2014; 124:3897

Summers C, Milano F, Gooley TA, Dahlberg A, Delaney C. Infusion of Ex Vivo Expanded Cord Blood Progenitor Cells Reduces the Risk of Bacteremia after Myeloablative Cord Blood Transplant. Blood 2014; 124:3860

Wagner JE, Eapen M, Carter S, Wang Y, Schultz KR, Wall DA, Bunin N, Delaney C, Haut P, Margolis D, Peres E, Verneris MR, Walters M, Horowitz MM, Kurtzberg J; Blood and Marrow Transplant Clinical Trials Network. One-unit versus two-unit cord-blood transplantation for hematologic cancers. N Engl J Med. 2014 Oct 30;371(18):1685-94.

Milano F, Shulman HM, Guthrie KA, Riffkin I, McDonald GB, Delaney C. Late-Onset Colitis after Cord Blood Transplantation is Consistent with Graft-versus-Host Disease: Results of a Blinded Histopathological Review. Biol Blood Marrow Transplant. 2014 Jul;20(7):1008-13.

Bar M, Wood BL, Radich JP, Doney KC, Woolfrey AE, Delaney C, Appelbaum FR, Gooley TA, Impact of minimal residual disease, detected by flow cytometry, on outcome of myeloablative hematopoietic cell transplantation for acute lymphoblastic leukemia. Leuk Res Treatment 2014; 2014:421723.

Delaney, C., Milano, F., Nicoud, I., Heimfeld, S., Karanes, C., Gutman, J., Wagner, J.; Appelbaum, F.R.. Bernstein, I. D.  Dose Dependent Enhancement Of Neutrophil Recovery By Infusion Of Notch Ligand Ex Vivo Expanded Cord Blood Progenitors: Results Of a Multi-Center Phase I Trial. Blood 2013; 122:297

Newell LF, Flowers MED, Gooley TA, Milano F, Carpenter PA, Martin PJ, Delaney C. Characteristics of Chronic GVHD after Cord Blood Transplantation. Bone Marrow Transplant. 2013 Oct;48(10):1285-90.

Ostronoff F, Milano F, Gooley T, Gutman JA, McSweeney P, Petersen FB, Sandmaier BM, Storb R, Delaney C. Double Umbilical Cord Blood Transplantation in Patients with Hematologic Malignancies Using a Reduced-Intensity Preparative Regimen without Anti-Thymocyte Globulin. Bone Marrow Transplant. 2013 Jun;48(6):782-6.

Milano F, Nelson L, Delaney C. Fetal maternal immunity and antileukemia activity in cord-blood transplant recipients. Bone Marrow Transplant. 2013 Mar;48(3):321-2.

Milano F, Heimfeld S, Gooley T, Jinneman J, Nicoud I, Delaney C. Correlation of Infused CD3(+)CD8(+) Cells with Single Donor Dominance after Double-Unit Cord Blood Transplantation. Biol Blood Marrow Transplant 2013 Jan: 19(1):156-60.

Delaney C, Becker PS, Milano F, Nicoud IB, Heimfeld S, Riffkin I, Papermaster A, Appelbaum FR, Bernstein ID, Estey EH. Infusion of “Off-the-Shelf” Third Party Ex Vivo Expanded Cord Blood Progenitor Cells as Supportive Care Following Clofarabine with High Dose Cytarabine and Granulocyte Colony-Stimulating Factor Priming for the Treatment of AML Blood 2011;118:3640

Milano F, Chien JW, Riffkin I, Gutman JA, Newell L, Pergam SA, Delaney C. Stable Long-Term Pulmonary Function after Myeloablative Double Cord Blood Transplant. BBMT, 2012 Feb;18(2):309-13.

Newell LF, Milano F, Nicoud IB, Pereira A, Gooley TA, Heimfeld S, Delaney C. Early CD3 peripheral blood chimerism predicts the long-term engrafting unit following myeloablative double cord blood transplantation. BBMT. 2012 Aug;18(8):1243-9.

Delaney C, Becker PS, Milano F, Nicoud IB, Heimfeld S, Riffkin I, Papermaster A, Appelbaum FR, Bernstein ID, Estey EH. Infusion of “off-the-shelf” third party ex vivo expanded cord blood progenitor cells as supportive care following clofarabine with high dose cytarabine and granulocyte colony-stimulating factor priming for the treatment of AML. Blood 2011;118:3640

Dahlberg A, Milano F, Gooley TA, Delaney C. The Risk of Day 100 Mortality Following Umbilical Cord Blood Transplantation is Significantly Higher in Patients Who do not Achieve an ANC of 100 by Day +16 Post Transplant Blood 2011; 118:3033

Brunstein C, Gutman J, Weisdorf C, Woolfrey A, DeFor T, Gooley T, Verneris M, Appelbaum F, Wagner J, Delaney C. Allogeneic Hematopoietic Cell Transplantation for Hematolotic Malignancy:Relative Risks and Benefirs of Double Umbilical Cord Blood. Blood 2010 Nov 25:(116922):4693-9.

Delaney C, Heimfeld S,Brashem-Stein C, Voorhies H, Manger R and Bernstein ID. Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nature Medicine, 2010 Feb;16(2):232-6.

Gutman JA, Turtle CJ, Manley TJ, Heimfeld S, Bernstein ID, Riddell SR, Delaney C. Single-unit Dominance after Double-Unit Umbilical Cord Blood Transplantation Coincides with a Specific CD8+ T-Cell Response Against theNonengrafted Unit (Plenary Paper) Bood 2010 Jan 28;115(4):757-65.

Delaney C, Gutman JA, Appelbaum FR. Cord Blood Transplant for Hematologic Malignancies: Conditioning Regimens, Double Cord Blood Transplant and Infectious Complications. British Journal of Hematology, 2009 Oct; 147(2):207-16.

Gutman JA, Leisenring W, Appelbaum FR, Woolfrey AE, Delaney C. Low relapse without excessive transplant related mortality following myeloablative cord blood transplantation for acute leukemia in complete remission: a matched cohort analysis. Biol Blood Marrow Transplant. 2009 Sep;15(9):1122-9.

Summers C, Grier A, Gardner R, Delaney C, Jensen M. Multiplexed Engineering of CD19CAR T Cells for Post-Transplant Consolidative Immunotherapy. Blood 2016; 128:1159

Milano, F., Merriam, F., Nicoud, I., Li, J., Gooley, T. A., Heimfeld, S., Imren, S., Delaney, C. Notch-Expanded Murine Hematopoietic Stem and Progenitor Cells Mitigate Death From Lethal Radiation and Convey Immune Tolerance in Mismatched Recipients. Stem Cells Transl Med. sctm.2016-0112

Dahlberg A, Woo S, Delaney C, Boyle P, Gnirke A, Bock C, Bernstein BE, Meissner A, Gottardo R, Bernstein ID. Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity. Leukemia. 2015 Sep; 29(9):1948-51

Hadland BK, Varnum-Finney B, Poulos MG, Moon RT, Butler JM, Rafii S, Bernstein ID. Endothelium and NOTCH specify and amplify aorta-gonad-mesonephros-derived hematopoietic stem cells. J Clin Invest. 2015 May; 125(5):2032-45.

Dahlberg A, Brasheim-Stein C, Delaney C, Bernstein ID. Enhanced Generation of Cord Blood Hematopoietic Stem and Progenitor Cells by Culture with Stem Regenin1 and Delta1(ext-IgG). Leukemia 2014 Oct 28 (10):2097-101.

Merriam FV, Nicoud I, Milano F, Li J, Heimfeld S, Delaney C. Notch-Expanded Murine Hematopoietic Progenitor Cells Can Induce Immune Tolerance in Mismatched Recipients. Blood 2014; 124:2430

Csaszar E, Wang W, Usenko T, Qiao W, Delaney C, Bernstein ID, Zandstra PW. Blood stem cell fate regulation by Delta-1-mediated rewiring of IL-6 paracrine signaling. Blood. 2014 Jan 30;123(5):650-8.

Gardner R, Summers C, Weber D, Jensen MC, Delaney C. Lentiviral Transduction Of Notch Ligand Expanded Cord Blood HSC’s To Express a CD19-Specific CAR Generates NK and Myeloid Progeny With Antitumor Activity. Blood 2013; 122:900

Watts KL , Delaney C, Nelson B, Trobridge GD, Beard BC, Humphries RK, Kiem HP. CD34+ Expansion with Delta1 and HOXB4 Promotes Rapid Engraftment and Transfusion Independence in a Macaca nemestrina Cord Blood Transplant Model. Mol Ther. 2013 Jun;21(6):1270-8.

Blake JM, Nicoud IB, Weber D,Voorhies H, Guthrie KA, Heimfeld S, Delaney C. Improved immunomagnetic enrichment of CD34 (+) cells from umbilical cord blood using the CliniMACS cell separation system. Cytotherapy. 2012 Aug; 14(7):818-22.

Bernstein I, Delaney C. Engineering Stem Cell Expansion. Cell Stem Cell. 10, 113 (2012).

Dahlberg A, Delaney C, Bernstein ID. Ex Vivo Expansion of Human Hematopoietic Stem and Progenitor Cells. Blood, 2011 Jun 9; 117(23):6083-90.

Varnum-Finney B, Halasz LM, Sun M, Gridley T, Radtke F, Bernstein ID. Notch2 governs the rate of generation of mouse long- and short-term repopulating stem cells. J Clin Invest. 2011 Mar;121(3):1207-16.

Varnum-Finney B,Dallas MH, Kato K, Bernstein ID. Notch target Hes5 ensures appropriate Notch induced T- versus B-cell choices in the thymus. Blood. 2008 Mar 1;111(5):2615-20.

Dallas MH, Varnum-Finney B, Martin PJ, Bernstein ID. Enhanced T-cell reconstitution by hematopoietic progenitors expanded ex vivo using the Notch ligand Delta1 Blood. 2007 Apr 15;109(8):3579-87.

Aoyama K, Delaney C,Varnum-Finney B, Kohn AD, Moon RT, Bernstein ID. The interaction of the Wnt and Notch pathways modulates natural killer versus T cell differentiation. Stem Cells. 2007 Oct; 25(10):2488-97.

Delaney C, Varnum-Finney B,Aoyama K, Brashem-Stein C, Bernstein ID. Dose-dependent effects of the Notch ligand Delta1 on marrow repopulating ability of cord blood cells. Blood. 106(8):2693-9, Oct 2005.

Dallas MH, Varnum-Finney B, Delaney C, Kato K, Bernstein ID. Density of the Notch ligand Delta1 determines generation of B and T cell precursors from hematopoietic stem cells. J Exp Med. 201(9):1361-6, 2005

Varnum-Finney B,Brashem-Stein C, Bernstein ID. Combined effects of Notch signaling and cytokines induce a multiple log increase in precursors with lymphoid and myeloid reconstituting ability. Blood, 101: 1784-1789, 2003.

Ohishi K1, Varnum-Finney B, Bernstein ID. Delta-1 enhances marrow and thymus repopulating ability of human CD34(+) CD38(-) cordblood cells. J Clin Invest. 2002 Oct;110(8):1165-74.

Ohishi K1, Varnum-Finney B, Serda RE, Anasetti C, Bernstein ID. The Notch ligand, Delta-1, inhibits the differentiation of monocytes into macrophages but permits their differentiation into dendritic cells. Blood. 2001 Sep 1;98(5):1402-7.

Ohishi K1, Varnum-Finney B, Flowers D, Anasetti C, Myerson D, Bernstein ID. Monocytes express high amounts of Notch and undergo cytokine specific apoptosis following interaction with the Notch ligand, Delta-1. Blood. 2000May 1;95(9):2847-54.

Varnum-Finney B, Wu L,Yu M, Brashem-Stein C, Staats S, Flowers D, Griffin JD, Bernstein ID. Immobilization of Notch ligand Delta-1 is required for induction of Notch signaling. J Cell Sci 2000 113:4313-4318.

Varnum-Finney B, Xu L, Brashem-Stein C, Nourigat C, Flowers D, Bakkour S,Pear WS, and Bernstein ID. Pluripotent, cytokine dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling. Nature Med6:1278-1281, 2000.

Varnum-Finney B, Purton LE, Yu M, Brashem-Stein C, Flowers D, Staats S, Moore KA, Le Roux I, Mann R, Gray G, Artavanis-Tsakonas S, Bernstein ID. The Notch ligand,Jagged-1, influences the development of primitive hematopoietic precursor cells. Blood. 1998 Jun 1;91(11):4084-91.

Milner LA, Kopan R, Martin DI, Bernstein ID. A human homologue of the Drosophila developmental gene, Notch, is expressed in CD34+ hematopoietic precursors. Blood. 1994 Apr 15;83(8):2057-62.