[Show abstract][Hide abstract] ABSTRACT: Tumor-specific adenoviral vectors comprise a fruitful gene-based diagnostic imaging and therapy research area for advanced stage of cancer, including metastatic disease. However, clinical translation of viral vectors has encountered considerable obstacles, largely due to host immune responses against the virus. Here, we explored the utilization of an immunosuppressant, rapamycin, to circumvent the anti-adenovirus immunity in immunocompetent murine prostate cancer models. Rapamycin diminished adenoviral-induced acute immune response by inhibiting NF-κB activation; it also reduced the scale and delayed the onset of inflammatory cytokine secretion. Further, we found that rapamycin abrogated anti-adenovirus antibody production and retarded the function of myeloid cells and lymphocytes that were activated upon viral administration in pre-immunized hosts. Thus, the co-administration of rapamycin prolonged and enhanced adenovirus-delivered transgene expression in vivo, and thereby augmented the imaging capability of adenoviral vectors in both bioluminescent and positron emission tomography modalities. Furthermore, we showed that despite an excellent response of cancer cells to a cytotoxic gene therapeutic vector in vitro, only minimal therapeutic effects were observed in vivo in pre-immunized mice. However, when we combined gene therapy with transient immunosuppression, complete tumor growth arrest was achieved. Overall, transient immunosuppression by rapamycin was able to boost the diagnostic utility and therapeutic potentials of adenoviral vectors.
PLoS ONE 09/2013; 8(9):e73650. DOI:10.1371/journal.pone.0073650 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We sought to modify adenoviral (Ad) particles by incorporating the advantageous characteristics of non-viral gene delivery vehicles to complement the viral vectors. α-Amino acid-N-carboxyanhydride chemistry was used to synthesize homopolypeptides and diblock copolypeptides that possess well-defined secondary structures. Using cryo-electron and fluorescence microscopy, we showed that these polypeptides can coat the surfaces of Ad particles in a non-covalent manner to modify their transduction properties. The coated Ad particles were found to bind to and be internalized by cells. In contrast to reports using covalently PEGylated Ad particles, we found that our physically coated Ad hybrid complexes facilitate gene transfer both in vitro and in vivo. We showed that our polypeptide coating was able to shield the Ad particles from the neutralizing effect of antibodies and mitigate the binding of blood coagulation factor (Factor X) in vitro. The coating also reduced the antigenicity of Ad in immunocompetent mice. The biodistribution of the systemically administered hybrid complexes mirrored the behavior of both viral and non-viral vectors, exhibiting liver tropism as well as enhanced lung transduction. These data demonstrated that our non-covalent modification was able to alter Ad's interactions with cells and organs with retention of transduction efficiency. Advantages such as facile coating of the Ad vector, design flexibility and ease of attaching ligands to the polypeptides make this system potentially useful as a platform for adding functionalities to Ad to target cancer metastasis.
[Show abstract][Hide abstract] ABSTRACT: Better intraprostatic cancer imaging techniques are needed to guide clinicians in prostate cancer treatment decisions. Because many genes are specifically overexpressed in cancer cells, one strategy to improve prostate cancer detection is to image intraprostatic cancer-specific transcriptional activity. Because of the obstacles of weak cancer- or tissue-specific promoter activity and bladder clearance of many PET tracers, intraprostatic PET of gene transcriptional activity has not been previously reported.
The two-step transcriptional amplification (TSTA) system that amplifies the prostate-specific antigen promoter activity was used for PET imaging of the reporter gene herpes simplex virus type-1 sr39 thymidine kinase (HSV1-sr39tk). The TSTA-sr39tk system was injected directly into prostates or prostatic tumors as a replication-incompetent adenovirus (AdTSTA-sr39tk) and imaged using PET.
AdTSTA-sr39tk was able to image prostate-specific antigen promoter transcriptional activity by 9-(4-(18)F-fluoro-3-[hydroxymethyl]butyl)guanine PET, in both mouse and canine prostates in vivo. Ex vivo small-animal PET images, scintigraphic counts, and sr39tk expression analysis confirmed the specificity of the observed signal.
Here, by combining the TSTA-amplified signal with a protocol for tracer administration, we show that in vivo PET detection of transcriptional activity is possible in both mouse and immunocompetent canine prostates. These results suggest that imaging applications using transcription-based tumor-specific promoters should be pursued to better visualize cancer foci that escape detection by conventional biopsies.
Journal of Nuclear Medicine 05/2011; 52(5):784-91. DOI:10.2967/jnumed.110.084582 · 6.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Tumor-infiltrating myeloid cells (TIMs) sup-port tumor growth by promoting angiogen-esis and suppressing antitumor immune responses. CSF-1 receptor (CSF1R) signal-ing is important for the recruitment of CD11b F4/80 tumor-associated macro-phages (TAMs) and contributes to myeloid cell-mediated angiogenesis. However, the impact of the CSF1R signaling pathway on other TIM subsets, including CD11b Gr-1 myeloid-derived suppressor cells (MDSCs), is unknown. Tumor-infiltrating MDSCs have also been shown to contribute to tumor angiogenesis and have recently been impli-cated in tumor resistance to antiangiogenic therapy, yet their precise involvement in these processes is not well understood. Here, we use the selective pharmacologic inhibitor of CSF1R signaling, GW2580, to demonstrate that CSF-1 regulates the tumor recruitment of CD11b Gr-1 lo Ly6C hi mono-nuclear MDSCs. Targeting these TIM subsets inhibits tumor angiogenesis associated with reduced expression of proangiogenic and immunosuppressive genes. Combination therapy using GW2580 with an anti– VEGFR-2 antibody synergistically sup-presses tumor growth and severely im-pairs tumor angiogenesis along with reverting at least one TIM-mediated anti-angiogenic compensatory mechanism in-volving MMP-9. These data highlight the importance of CSF1R signaling in the recruitment and function of distinct TIM subsets, including MDSCs, and validate the benefits of targeting CSF1R signaling in combination with antiangiogenic drugs for the treatment of solid cancers. (Blood.
[Show abstract][Hide abstract] ABSTRACT: Tumor-infiltrating myeloid cells (TIMs) support tumor growth by promoting angiogenesis and suppressing antitumor immune responses. CSF-1 receptor (CSF1R) signaling is important for the recruitment of CD11b(+)F4/80(+) tumor-associated macrophages (TAMs) and contributes to myeloid cell-mediated angiogenesis. However, the impact of the CSF1R signaling pathway on other TIM subsets, including CD11b(+)Gr-1(+) myeloid-derived suppressor cells (MDSCs), is unknown. Tumor-infiltrating MDSCs have also been shown to contribute to tumor angiogenesis and have recently been implicated in tumor resistance to antiangiogenic therapy, yet their precise involvement in these processes is not well understood. Here, we use the selective pharmacologic inhibitor of CSF1R signaling, GW2580, to demonstrate that CSF-1 regulates the tumor recruitment of CD11b(+)Gr-1(lo)Ly6C(hi) mononuclear MDSCs. Targeting these TIM subsets inhibits tumor angiogenesis associated with reduced expression of proangiogenic and immunosuppressive genes. Combination therapy using GW2580 with an anti-VEGFR-2 antibody synergistically suppresses tumor growth and severely impairs tumor angiogenesis along with reverting at least one TIM-mediated antiangiogenic compensatory mechanism involving MMP-9. These data highlight the importance of CSF1R signaling in the recruitment and function of distinct TIM subsets, including MDSCs, and validate the benefits of targeting CSF1R signaling in combination with antiangiogenic drugs for the treatment of solid cancers.
[Show abstract][Hide abstract] ABSTRACT: Imaging in medicine has been classically based on the anatomical description of organs. In the past 15 years, new imaging techniques based on gene expression that characterize a pathological process have been developed. Molecular imaging is the use of such molecules to image cell-specific characteristics. Here, we review recent advances in molecular imaging, taking as our prime example lymph node (LN) metastasis in prostate cancer. We describe the new techniques and compare their accuracy in detecting LN metastasis in prostate cancer. We also present new molecular strategies for improving tumor detection using adenoviruses, molecular promoters and amplification systems. Finally, we present the concept of 'in vivo pathology', which envisages using molecular imaging to accurately localize metastatic lesions based on the molecular signature of the disease.
Trends in Molecular Medicine 07/2009; 15(6):254-62. DOI:10.1016/j.molmed.2009.04.004 · 9.45 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Because of its high selectivity and specificity for the imaging reporter probe 9-(4-(18)F-fluoro-3-[hydroxymethyl]butyl)guanine ((18)F-FHBG), the herpes simplex virus type 1 thymidine kinase (HSV1-tk) variant sr39tk is actively being studied as a PET reporter gene. We recently demonstrated the capability of using a prostate-specific transcriptional amplification PET reporter vector, AdTSTA-sr39tk, to target prostate cancer lymph node metastasis. However, one area that warrants further study is the examination of the sensitivity of PET by determining the minimum percentage of cells expressing the sr39tk transgene needed for detection. Addressing this question could determine the sensitivity of vector-mediated sr39tk PET in cancer-targeting strategies.
DU-145, PC-3, and CWR22Rv.1 prostate cancer cell lines (a total of 1 x 10(6) cells) were studied, of which 7%, 10%, 25%, 50%, or 70% were transduced with the lentiviral vector constitutively expressing HSV1-sr39tk-IRES-enhanced green fluorescent protein (EGFP). Cells were subcutaneously implanted into the left shoulder of severe combined immunodeficient mice and evaluated. Tumor cells comparably transduced with an EGFP control vector were implanted on the right shoulder. Mice were imaged using PET with (18)F-FHBG at 8, 15, and 22 d after tumor implant. On day 23, tumors were isolated and analyzed for sr39tk transgene expression by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR), Western blotting, immunohistochemistry, and flow cytometry for EGFP expression.
Results showed a linear relationship between the level of sr39tk expression and the quantity of tracer accrual in DU-145, with the minimal value for PET detection at 10%. The magnitude of tracer retention in sr39tk-expressing cells was amplified over time as the tumor grew. Protein levels in the stepwise titration increased with the percentage of sr39tk-transduced cells.
The stepwise titration of prostate cancer cells transduced with the lenti-CMV-sr39tk-IRES-EGFP determined the minimum number of sr39tk-expressing tumor cells necessary to be detected by PET using the (18)F-FHBG reporter probe. Furthermore, PET signal correlated well with traditional methods of protein evaluation such as flow cytometry, quantitative RT-PCR, Western blotting, and immunohistochemistry. Unlike the traditional methods, however, the use of PET is noninvasive and will be more advantageous in clinical situations.
Journal of Nuclear Medicine 05/2009; 50(5):757-64. DOI:10.2967/jnumed.108.058438 · 6.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The accurate assessment of nodal involvement in prostate cancer is crucial to planning treatment, yet there is a shortage of noninvasive imaging techniques capable of visualizing nodal lesions directly. This study demonstrates the feasibility of using recombinant human adenoviral vectors to detect nodal metastases in a human prostate cancer model. This was achieved by the prostate-restricted expression of optical and positron emission tomography (PET) imaging reporter genes by the viral vector coupled with the innate lymphotropic properties of adenovirus. We show that peritumoral administration of these vectors results in the direct detection of reporter gene expression in metastatic lesions within sentinel lymph nodes. Notably, this approach parallels the current lymphoscintigraphy method but enables the direct PET visualization of sentinel lymph node metastases, eliminating the need for invasive lymphadenectomy. These findings may lead to more effective diagnostic and therapeutic strategies for individuals with advanced-stage prostate cancer.
Nature medicine 08/2008; 14(8):882-8. DOI:10.1038/nm.1727 · 27.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Prostate cancer dissemination is difficult to detect in the clinic, and few treatment options exist for patients with advanced-stage disease. Our aim was to investigate the role of tumor lymphangiogenesis during metastasis. Further, we implemented a noninvasive molecular imaging technique to facilitate the assessment of the metastatic process. The metastatic potentials of several human prostate cancer xenograft models, LAPC-4, LAPC-9, PC3 and CWR22Rv-1 were compared. The cells were labeled with luciferase, a bioluminescence imaging reporter gene, to enable optical imaging. After tumor implantation the animals were examined weekly during several months for the appearance of metastases. Metastatic lesions were confirmed by immunohistochemistry. Additionally, the angiogenic and lymphangiogenic profiles of the tumors were characterized. To confirm the role of lymphangiogenesis in mediating metastasis, the low-metastatic LAPC-9 tumor cells were engineered to overexpress VEGF-C, and the development of metastases was evaluated. Our results show CWR22Rv-1 and PC3 tumor cell lines to be more metastatic than LAPC-4, which in turn disseminates more readily than LAPC-9. The difference in metastatic potential correlated with the endogenous production levels of lymphangiogenic growth factor VEGF-C and the presence of tumor lymphatics. In agreement, induced overexpression of VEGF-C in LAPC-9 enhanced tumor lymphangiogenesis leading to the development of metastatic lesions. Taken together, our studies, based on a molecular imaging approach for semiquantitative detection of micrometastases, point to an important role of tumor lymphatics in the metastatic process of human prostate cancer. In particular, VEGF-C seems to play a key role in prostate cancer metastasis.
International Journal of Cancer 11/2007; 121(10):2153-61. DOI:10.1002/ijc.22900 · 5.09 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A better understanding of the in vivo biodistribution of adenoviral vectors would enable the researcher to anticipate potential side effects due to off-targeted site of transduction, and aid in the strategic design of gene therapy. We combined real-time polymerase chain reaction with in vivo optical imaging to examine viral transduction in liver, lung, spleen, kidney, prostate, and lymph nodes. A replication-deficient serotype 5 adenoviral vector expressing the firefly luciferase gene under the control of a constitutive cytomegalovirus promoter was administered in vivo via different routes. Intravenous and intraperitoneal injections resulted in greatest gene expression and viral DNA in the liver, whereas intraperitoneal injections led to a greater extent of gene delivery to the prostate. Although prostate-directed injection resulted in dominant gene expression in the targeted site, leakage of the vector to other organs was also observed. Vector injection into the lymphatic-rich paw tissue or the subcutaneous tissue of shoulder or chest followed the expected lymphatic drainage pattern, resulting in the accumulation of viral vector in ipsilateral brachial and axillary lymph nodes. Collectively, this study demonstrates that each tissue retains various amounts of adenoviral vector, depending on the route of administration. This knowledge is useful in the strategic design and implementation of adenovirus-mediated gene therapies.
Human Gene Therapy 01/2007; 17(12):1262-9. DOI:10.1089/hum.2006.17.1262 · 3.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Significant advances in gene therapy have been made as a result of the improvement of gene delivery systems, discovery of new therapeutic genes, better understanding of mechanisms of disease progression, exploration and improvement of tissue-specific gene regulatory sequences, and development of better prodrug/enzyme systems. This review discusses adenoviral-based and prostate-specific cancer gene therapy--emphasizing tissue-specific promoter choices to increase gene therapy safety and specificity--and the development of prostate-targeted vectors, with a focus on the two-step transactivation system for amplifying gene expression, specifically in prostate cancer cells. Several examples will be discussed for the scientific basis and therapeutic applications. In addition, prostate cancer gene therapy clinical trials and future directions in this field will also be described briefly.
[Show abstract][Hide abstract] ABSTRACT: Gene-based therapy is a promising and flexible therapeutic approach to manage diverse types of cancer. The lack of convincing therapeutic success of current gene therapy protocols in part, can be attributed to the inability to monitor gene expression at the targeted site in the living subject. Linking molecular imaging to gene therapy will enable real-time assessment of the therapeutic process and the refinement of treatment protocols. This review will cover two common imaging modalities, positron emission tomography (PET) and bioluminescence imaging (BLI), used in pre-clinical and clinical gene therapy applications. Strategies to develop more specific and robust cancer gene therapy and imaging approaches will be discussed. Coupling PET to gene therapy of cancer has already been implemented in several clinical studies. This approach would help to improve the efficacy and safety of future gene therapy clinical trials.
Current Gene Therapy 01/2006; 5(6):607-18. DOI:10.2174/156652305774964695 · 2.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Cancer gene therapy based on tissue-restricted expression of cytotoxic gene should achieve superior therapeutic index over an unrestricted method. This study compared the therapeutic effects of a highly augmented, prostate-specific gene expression method to a strong constitutive promoter-driven approach. Molecular imaging was coupled to gene therapy to ascertain real-time therapeutic activity. The imaging reporter gene (luciferase) and the cytotoxic gene (herpes simplex thymidine kinase) were delivered by adenoviral vectors injected directly into human prostate tumors grafted in SCID mice. Serial bioluminescence imaging, positron emission tomography (PET) and computed tomography (CT) revealed restriction of gene expression to the tumors when prostate-specific vector was employed. In contrast, administration of constitutive active vector resulted in strong signals in the liver. Liver serology, tissue histology, and frail condition of animals confirmed liver toxicity suffered by the constitutive active cohorts, while the prostate-targeted group was unaffected. The extent of tumor killing was analyzed by apoptotic staining and human prostate marker (PSA). Overall, the augmented prostate-specific expression system was superior to the constitutive approach in safeguarding against systemic toxicity, while achieving effective tumor-killing. Integrating non-invasive imaging into cytotoxic gene therapy will provide a useful strategy to monitor gene expression and therapeutic efficacy in future clinical protocols.
[Show abstract][Hide abstract] ABSTRACT: Due to the exceptionally limited treatment options currently available for recurrent hormone refractory prostate cancer (HRPC), gene therapy can provide a tool to fight this Herculean disease. To develop a safe and efficient vector-based gene delivery approach, we employed a prostate-targeted gene expression system. The combination of a modified PSA promoter/enhancer and two-step transcriptional amplification (TSTA) system enabled us to achieve robust expression of transgenes in our vectors while maintaining prostate specificity. Furthermore, integration of an efficient adenovirus-mediated gene delivery approach and noninvasive optical imaging allowed us to continuously monitor and evaluate our system on human prostate cancer mouse xenografted models (Sato, et al. Mol Ther, '03). We have also shown that the TSTA system can be usable for clinically-translatable mPET radionuclide imaging, applicable to HRPC where androgen receptor (AR) is expressed (Sato, et al. Clin Cancer Res, '05), and that the suicide gene therapy approach with the TSTA system exhibited promising results, as well as remarkably lower liver toxicity compared to conventional CMV promoter (Johnson et al. submitted).To further explore therapeutic approaches to utilize the TSTA system, we have taken an oncolytic approach to eradicate prostate cancer. Previously we had confirmed that two distinct genes can be expressed from a single construct because of the structural nature of the TSTA system. A pair of firefly luciferase (FL) and HSV-1 sr39tk genes flanking five repeats of GAL4 binding sites were both expressed from a single adenovirus vector simultaneously. We successfully obtained both optical and mPET imaging on the same animal by this bidirectional vector. Based on this, we developed an oncolytic adenovirus utilizing the TSTA system. In this new vector, both E1A and E1B driven by GAL4 binding sites will make the virus replication competent by the GAL4VP16 activator expressed in trans. Therefore, where the TSTA is active, E1A and E1B will be expressed, and this should lead to oncolytic adenoviral replication. In a preliminary study, virus replication was suggested by the increase of the FL activity from the AdTSTA-FL co-infected in the AR+ prostate cell line, LNCaP, following the transfection of the plasmid with E1A and E1B flanking two GAL4 repeats (G2). However, no increase of the FL activity was seen with the E1A and E1B plasmid with no GAL4 repeat (G0), possibly because of no virus production. The viral production was actually confirmed by a plaque formation in the transfection study with a G2 or G4 driving E1 proteins but was not for G0 in addition to the expression of E1 proteins. Viral replication was also confirmed by co-infection study using G4 adenovirus but not with G0 adenovirus. Currently we are working on analyzing G0, G2, and G4 adenovirus to see oncolytic activity and constructing all-in-one TSTA oncolytic adenoviruses. We foresee that this oncolytic approach based on the TSTA system can be another promising gene therapy strategy for the advanced stages of prostate cancer.
[Show abstract][Hide abstract] ABSTRACT: A highly augmented, prostate-specific two-step transcriptional amplification (TSTA) method was developed with the ultimate goal of delivering an effective and safe gene-based treatment to prostate cancer patients. Because very limited treatment options are available for recurrent hormone refractory prostate cancer (HRPC), it is imperative to assess whether the prostate-specific antigen (PSA) promoter-based TSTA gene therapy will be functional in HRPC.
We tested the TSTA-driven adenovirus vector on three androgen-dependent and six HRPC models. Real-time gene expression was monitored by both optical imaging and the combined modality of positron emission tomography (PET) and computed tomography.
The TSTA-driven firefly luciferase expressing adenoviral vector was active in all androgen receptor (AR)-expressing HRPC models, but inactive in AR- and PSA-negative lines. Interestingly, the TSTA-mediated gene expression was induced by hydrocortisone in MDA PCa 2b, a cell line with mutated AR that possesses altered ligand specificity. In animal models, the TSTA-mediated optical signal was more robust in the HRPC than androgen-dependent tumors. In a parallel trend, a TSTA vector that expresses the herpes simplex virus thymidine kinase PET reporter gene also displayed more robust PET signal in the HRPC tumor.
The activity of TSTA system is AR dependent and it recapitulates the functional status of endogenous AR. These data support the conclusion that AR function is activated in HRPC despite castrated levels of androgen. Together with the fact that majority of recurrent prostate cancers express AR and PSA, we foresee that the TSTA approach can be a promising gene therapy strategy for the advanced stages of prostate cancer.
Clinical Cancer Research 06/2005; 11(10):3743-9. DOI:10.1158/1078-0432.CCR-04-1990 · 8.72 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Targeted gene transduction to specific tissues and organs through intravenous injection would be the ultimate preferred method of gene delivery. Here, we report successful targeting in a living animal through intravenous injection of a lentiviral vector pseudotyped with a modified chimeric Sindbis virus envelope (termed m168). m168 pseudotypes have high titer and high targeting specificity and, unlike other retroviral pseudotypes, have low nonspecific infectivity in liver and spleen. A mouse cancer model of metastatic melanoma was used to test intravenous targeting with m168. Human P-glycoprotein was ectopically expressed on the surface of melanoma cells and targeted by the m168 pseudotyped lentiviral vector conjugated with antibody specific for P-glycoprotein. m168 pseudotypes successfully targeted metastatic melanoma cells growing in the lung after systemic administration by tail vein injection. Further development of this targeting technology should result in applications not only for cancers but also for genetic, infectious and immune diseases.
Nature Medicine 04/2005; 11(3):346-52. DOI:10.1038/nm1192 · 27.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Cancer gene therapy based on tissue-restricted expression of cytotoxic gene should achieve superior therapeutic index over an unrestricted method. This study compared the therapeutic effects of a highly augmented, prostate-specific gene expression method to a strong constitutive promoter-driven approach. Molecular imaging was coupled to gene therapy to ascertain real-time therapeutic activity. The imaging reporter gene (luciferase) and the cytotoxic gene (herpes simplex thymidine kinase) were delivered by adenoviral vectors injected directly into human prostate tumors grafted in SCID mice. Serial bioluminescence imaging, positron emission tomography, and computed tomography revealed restriction of gene expression to the tumors when prostate-specific vector was employed. In contrast, administration of constitutive active vector resulted in strong signals in the liver. Liver serology, tissue histology, and frail condition of animals confirmed liver toxicity suffered by the constitutive active cohorts, whereas the prostate-targeted group was unaffected. The extent of tumor killing was analyzed by apoptotic staining and human prostate marker (prostate-specific antigen). Overall, the augmented prostate-specific expression system was superior to the constitutive approach in safeguarding against systemic toxicity, while achieving effective tumor killing. Integrating noninvasive imaging into cytotoxic gene therapy will provide a useful strategy to monitor gene expression and therapeutic efficacy in future clinical protocols.
[Show abstract][Hide abstract] ABSTRACT: Molecular Therapy (2004) 9, S373–S373; doi: 10.1016/j.ymthe.2004.06.948
975. TSTA Mediated Multi-Modality Gene-Expression Imaging in Hormone Refractory Prostate Cancer
Makoto Sato1, Mai Johnson1, Liqun Zhang2, Sanjiv S. Gambhir3,4, Michael Carey2,3 and Lily Wu1,31Urology2Biochemistry3Crump Institute for Molecular Imaging, UCLA School of Medicine, Los Angeles, CA4Department of Radiology and Bio-X Program, Stanford Univ, Stanford, CA