Susan R. S. Gottesman

Transcription factors TFE3 and TFEB are critical for CD40 ligand expression and thymus-dependent humoral immunity

TFE3 and TFEB are broadly expressed transcription factors related to the transcription factor Mitf. Although they have been linked to cytokine signaling pathways in nonlymphoid cells, their function in T cells is unknown. TFE3-deficient mice are phenotypically normal, whereas TFEB deficiency causes early embryonic death. We now show that combined inactivation of TFE3 and TFEB in T cells resulted in a hyper–immunoglobulin M syndrome due to impaired expression of CD40 ligand by CD4+ T cells. Native TFE3 and TFEB bound to multiple cognate sites in the promoter of the gene encoding CD40 ligand (Cd40lg), and maximum Cd40lg promoter activity and gene expression required TFE3 or TFEB. Thus, TFE3 and TFEB are direct, physiological and mutually redundant activators of Cd40lg expression in activated CD4+ T cells critical for T cell–dependent antibody responses.

Peripheral T Cell Lymphoma in a Patient with Common Variable Immunodeficiency Disease: Case Report and Literature Review

This report documents the occurrence of a peripheral T cell lymphoma arising in the bone marrow and liver of a patient with common variable immunodeficiency disease. The T cell origin of this lymphoma was demonstrated by immunohistochemical phenotyping and gene rearrangement studies and was not associated with EBV infection of the lymphoma cells. The frequency and characteristics of lymphomas complicating CVID are reviewed.

High-grade T-cell lymphoma complicating B-cell chronic lymphocytic leukemia: An unusual manifestation of “Richter's syndrome”

Approximately 3% of patients with B‐cell chronic lymphocytic leukemia (CLL) develop a high‐grade large‐cell lymphoma consistent with Richters Syndrome. In most cases, these lymphomas are of B‐cell origin and are believed to arise by clonal evolution from the CLL cells. We present a case of a patient with a 10‐year history of B‐CLL who developed an aggressive large‐cell lymphoma, confirmed by immunophenotype to be of T‐cell origin. We suggest that in patients with CLL, immunodysregulation can result in the proliferation of T cells, which may mutate and result in the development of a new malignant clone. Am. J. Hematol. 66:203–206, 2001.

Plasma cell tumors in HIV-positive patients: Report of a case and review of the literature

Plasma cell tumors show an increased incidence in HIV-positive patients. The cases reported in the literature suggest that plasma cell tumors occur in a younger age group than that encountered in the general population. Pathologically, many of these tumors show a plasmablastic morphology. Plasma cell tumors in HIV-positive patients may present at unusual sites and progress rapidly to involve multiple sites, including the soft tissues and viscera. The prognosis is generally poor. These features may be related to a combination of factors, including immunodeficiency, oncogenic viruses, and altered cytokine milieu in these patients. A case of plasma cell tumor in an HIV-positive patient is presented.

T Cell Lymphoma Associated with Myelofibrosis

Myelofibrosis is most frequently associated with certain primary myeloproliferative disorders, but is rare in lymphoid neoplasms. We report the fourth case associated with T-cell lymphoma, involving bone marrow, lymph nodes and spleen. Marked extramedullary hematopoiesis was present. Myelofibrosis subsided completely with response to standard anti-lymphoma combination chemotherapy. Since lymphomatous splenic and bone marrow involvement was minimal in our patient, fibrotic bone marrows should be carefully evaluated for lymphoma.

Dual Inhibition of CDK4 and CDK2 via Targeting p27 Tyrosine Phosphorylation Induces a Potent and Durable Response in Breast Cancer Cells

Cyclin-dependent kinase 4/6 (CDK4/6)-specific inhibitors, such as palbociclib, have shown clinical efficacy, but primary or secondary resistance has emerged as a problem. To develop more effective therapeutic approaches, investigation is needed into the mechanisms of resistance or adaption. Here, it is demonstrated that CDK2 compensates for loss of CDK4 activity to rescue palbociclib-arrested breast cancer cells, suggesting that inhibition of both kinases is required to achieve durable response. In addition, a novel strategy is described to inhibit tyrosine phosphorylation of p27Kip1 (CDKN1B) and simultaneously inhibit both CDK2 and CDK4. p27Kip1 is a required assembly factor for cyclin–CDK4 complexes, but it must be phosphorylated on residue Y88 to open or activate the complex. The Brk-SH3 peptide, ALT, blocks p27 Y88 phosphorylation, inhibiting CDK4. Nonphosphorylated p27 is no longer a target for ubiquitin-mediated degradation and this stabilized p27 now also inhibits CDK2 activity. Thus, ALT induction inhibits both the kinase that drives proliferation (CDK4) and the kinase that mediates resistance (CDK2), causing a potent and long-lasting cell-cycle arrest. ALT arrests growth of all breast cancer subgroups and synergizes with palbociclib to increase cellular senescence and to cause tumor regression in breast cancer xenograft models. The use of ALT demonstrates that both CDK4 and CDK2 need to be inhibited if long-term efficacy is to be achieved and represents a novel modality to inhibit breast cancer cells. Implications: Modulating tyrosine phosphorylation of p27 impacts both proliferative (CDK4) and resistance (CDK2) mechanisms in breast cancer and suggests that phospho-p27 status may serve as a biomarker for patients that are responsive to CDK4/6 inhibition. Mol Cancer Res; 16(3); 361–77. ©2018 AACR.

Abstract 2348: Targeting the p27kip1/cdk4/cdk2/Rb axis in breast cancer using a peptidomimetic of Brk’s SH3 domain

Purpose: Cyclin D-cdk4 (DK4) has been a highly sought after therapeutic target because it drives cancer proliferation in a majority of human tumors. We have explored the clinical utility of a recently discovered mechanism of cell cycle control exerted on DK4 by p27Kip1 and its activator, the Breast tumor Related Kinase (Brk), in predicting responsiveness to therapy and as a new target for treatment. Although known as a DK4 assembly factor and cdk2 inhibitor, p27 also acts as a DK4 ON/OFF “switch.” Tyrosine (Y) phosphorylation of p27 (pY) by Brk gatekeeps both ATP binding and CAK phosphorylation of cdk4’s T loop, essential for DK4 activation. This function is restricted to cdk4: p27’s association with cdk2, whether Y phosphorylated or not, appears to be inhibitory. However, in vivo Y phosphorylated p27 is a target for cdk2-dependent ubiquitin-mediated degradation, reducing p27’s association with cdk2, indirectly activating this complex. We showed that blocking p27 pY inactivates cdk4 directly AND cdk2 indirectly, and thus represents a novel way to block cancer cell proliferation. pY also serves as a predictive biomarker of cdk4 activity and tumor response. Methods: We used a small peptide, ALT, which contains a portion of Brk’s SH3 domain. ALT binds to p27, blocks Brk’s association and ability to phosphorylate p27, inhibiting cdk4 and increasing p27’s ability to inhibit cdk2. We engineered a lipid-based nanoparticle delivery vehicle (NP-ALT), permitting us to test ALT as a first generation therapeutic in breast cancer cell lines that were both responsive and non-responsive to cdk4i therapy. ALT was also used with Palbociclib to determine if combination therapy reduced drug resistance. We developed a dual IHC assay for p27 and pY, which we used to analyze paraffin-embedded, archival human tumor samples, to determine whether we could pinpoint patients who would have responded to cdk4 inhibition therapy. Results: NP-ALT blocks pY, cdk4 and cdk2 activity, and proliferation in both Palbociclib sensitive and resistant cell lines. As a dual therapy, ALT treatment synergized with Palbociclib to arrest cells for >30 days, increased senescence, and in animal models caused tumor regression instead of just slowing tumor growth as seen with Palbociclib alone. Analysis of human cancer, obtained from archival sources, demonstrated that pY is never detected in quiescent benign mammary tissue, but is detected in about half of the advanced ER/PR+/Her2- tumors analyzed, and using explant culture techniques, we were able to stratify pY with Palbociclib response. Conclusion: Use of an Brk SH3 based peptide (NP-ALT) has proven effective in blocking p27 pY, inhibiting both cdk2 and cdk4, inducing senescence and increased durability. pY levels correlate with Palbociclib sensitivity in low, moderate and non-responders, suggesting that this may be a biomarker highlighting responsiveness to cdk4i therapy. Citation Format: Stacy W. Blain, Jason Quinones, Priyank Patel, Vladislav Tsiperson, Susan Gottesman, Jonathan Somma, Yun Wu. Targeting the p27kip1/cdk4/cdk2/Rb axis in breast cancer using a peptidomimetic of Brk’s SH3 domain [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2348. doi:10.1158/1538-7445.AM2017-2348